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  • Articles  (1,271)
  • Industrial Chemistry and Chemical Engineering  (1,133)
  • biofilm  (72)
  • continuous culture  (66)
  • Process Engineering, Biotechnology, Nutrition Technology  (1,271)
  • 1
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Bioresource Technology 47 (1994), S. 275-282 
    ISSN: 0960-8524
    Keywords: Anaerobic filter ; acetate ; biofilm ; biomass ; egg albumin ; glucose ; growth yield ; methane production ; two-phase process
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Process Engineering, Biotechnology, Nutrition Technology
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  • 2
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Bioresource Technology 48 (1994), S. 155-161 
    ISSN: 0960-8524
    Keywords: Effective diffusivity ; anaerobic digestion ; biofilm ; bioflocs ; fatty acids ; methane ; reaction-diffusion
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Process Engineering, Biotechnology, Nutrition Technology
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  • 3
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Enzyme and Microbial Technology 16 (1994), S. 1079-1083 
    ISSN: 0141-0229
    Keywords: Trichoderma reesei ; cellulase biosynthesis ; comparison with other mutants of T. reesei and soluble substrates ; continuous culture ; growth and maintenance parameters ; lactose
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 4
    Electronic Resource
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    Amsterdam : Elsevier
    Enzyme and Microbial Technology 16 (1994), S. 467-477 
    ISSN: 0141-0229
    Keywords: Catharanthus roseus ; Cinchona robusta ; Hydrodynamic stress ; Nicotiana tabacum ; Tabernaemontana divaricata ; batch culture ; continuous culture ; plant cells
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 5
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    Amsterdam : Elsevier
    Enzyme and Microbial Technology 15 (1993), S. 652-656 
    ISSN: 0141-0229
    Keywords: Dihydrofolate reductase ; Escherichia coli ; continuous culture ; methotrexate
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 6
    ISSN: 0141-0229
    Keywords: Curve fitting ; antibiotics ; continuous culture ; secondary metabolism
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 7
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    Springer
    Journal of industrial microbiology and biotechnology 15 (1995), S. 45-48 
    ISSN: 1476-5535
    Keywords: Kluyveromyces fragilis ; lactose transport ; continuous culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Lactose transport was studied inKluyveromyces fragilis grown in lactose-limited chemostat cultures. Kinetic parameters were determined using a method based on genetic population evolution. Lactose transport was carried out via three carriers characterized respectively byK m of 0.1 mM, 3 mM and 15.5 mM. The synthesis of these lactose carriers and their capacity (V max) are dependent on the dilution rate (D). At D=0.12 h−1, the high affinity transporter is prominent. For intermediate dilution rate, only the high and the medium affinity systems are present. In cells growing at D=0.4 h−1, these carriers are absent but instead, the low affinity transporter is present. The effect on lactose transport of such metabolic inhibitors as CCCP, a proton ionophore, and Antimycin A, an energy inhibitor, were also investigated. The high affinity system is the most sensitive to the effect of these inhibitors. Lactose transport through this carrier is probably a mechanism dependent on the proton motive force.
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  • 8
    ISSN: 1476-5535
    Keywords: S. epidermidis ; biofilm ; slime ; lectin marker
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A lectin-biotin assay was developed for use in the specific detection of slime produced byStaphylococcus epidermidis RP62A and M187sp11 grown in a chemically defined medium. Mature biofilm was formed on polyvinylchloride (PVC) disks using a combined chemostat-modified Robbins device (MRD) model system. Specimens fixedin situ were: 1) stained with ruthenium red; 2) reacted overnight with biotin-labeled lectins (WGA, succinyl-WGA, Con A, or APA) followed by treatment with gold-labeled extravidin; or 3) reacted with antibodies againstS. epidermidis RP62A capsular polysaccharide/adhesin (PS/A) using an immunogold procedure. WGA and succinyl-WGA (S-WGA), which specifically bindN-acetylglucosamine, were shown by TEM to react only with slime, both cell-associated and exocellular. In contrast, Con A, APA and anti-PS/A reacted with the bacterial cell surface but did not react with slime. These results indicate the usefulness of WGA lectin as a specific marker for detection of the presence and distribution of slime matrix material inS. epidermidis biofilm.
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  • 9
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    Springer
    Journal of industrial microbiology and biotechnology 16 (1996), S. 124-128 
    ISSN: 1476-5535
    Keywords: denitrification ; continuous culture ; oxygen ; Comamonas sp
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Continuous cultures ofComamonas sp SGLY2 were grown anaerobically prior to establishing steady states at different oxygen flow rates. At a low oxygen transfer rate, no dissolved oxygen accumulated in the medium and all nitrate was reduced to dinitrogen. Concurrently with the increase of dissolved oxygen concentration in the liquid phase, the rate of denitrification decreased. However, at a dissolved oxygen concentration near saturation (33 mg L−1), a part of the electron flow always diverted to nitrate with production of dinitrogen: the aerobic denitrification rate was equivalent to 35% of that calculated under anaerobic conditions. These experiments reflected the co-utilization of oxygen and N-oxides and the production of dinitrogen, up to saturated conditions, which implied synthesis and activity of the four denitrifying enzymes under various aeration conditions.
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  • 10
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    Springer
    Journal of industrial microbiology and biotechnology 16 (1996), S. 249-256 
    ISSN: 1476-5535
    Keywords: ethanol ; biofilm ; plastic composite-supports ; Zymomonas ; Saccharomyces
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Continuous ethanol fermentations were performed in duplicate for 60 days withZymomonas mobilis ATCC 331821 orSaccharomyces cerevisiae ATCC 24859 in packed-bed reactors with polypropylene or plastic composite-supports. The plastic composite-supports used contained polypropylene (75%) with ground soybean-hulls (20%) and zein (5%) forZ. mobilis, or with ground soybean-hulls (20%) and soybean flour (5%) forS. cerevisiae. Maximum ethanol productivities of 536 gL−1 h−1 (39% yield) and 499 gL−1 h−1 (37% yield) were obtained withZ. mobilis on polypropylene and plastic composite-supports of soybean hull-zein, respectively. ForZ. mobilis, and optimal yield of 50% was observed at a 1.92h−1 dilution rate for soybean hull-zein plastic composite-supports with a productivity of 96gL−1h−1, whereas with polypropylene-supports the yield was 32% and the productivity was 60gL−1h−1. With aS. cerevisiae fermentation, the ethanol production was less, with a maximum productivity of 76gL−1h−1 on the plastic composite-support at a 2.88h−1 dilution rate with a 45% yield. Polypropylene-support bioreactors were discontinued due to reactor plugging by the cell mass accumulation. Support shape (3-mm chips) was responsible for bioreactor plugging due to extensive biofilm development on the plastic composite-supports. With suspensionculture continuous fermentations in continuously-stirred benchtop fermentors, maximum productivities of 5gL−1h−1 were obtained with a yield of 24 and 26% withS. cerevisiae andZ. mobilis, respectively. Cell washout in suspensionculture continuous fermentations was observed at a 1.0h−1 dilution rate. Therefore, for continuous ethanol fermentations, biofilm reactors out-performed suspension-culture reactors, with 15 to 100-fold higher productivities (gL−1h−1) and with higher percentage yields forS. cerevisiae andZ. mobilis, respectively. Further research is needed with these novel supports to evaluate different support shapes and medium compositions that will permit medium flow, stimulate biofilm formation, reduce fermentation costs, and produce maximum yields and productivities.
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  • 11
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    Springer
    Journal of industrial microbiology and biotechnology 17 (1996), S. 228-234 
    ISSN: 1476-5535
    Keywords: colonization ; biofilm ; diversity ; proximal vertical packing ; cell-cell interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Using laminar flow chambers and time-lapse video imaging, colonization of surfaces by four marine bacteria revealed a diverse range of morphological characteristics and cell-cell interactions. The strain SW5 formed a compact, multilayered single- and double-cell biofilm on hydrophobic surfaces but developed long multicellular chains on hydrophilic surfaces. The morphologically similar SW8 showed unusual proximal vertical packing of cells on both substrata.Vibrio sp strain S14 exhibited cyclical colonization-detachment events on both substrata.Pseudomonas sp strain S9 initially displayed reversible and then irreversible adhesion apparently triggered by a cell density phenomenon that led to the development of regular microcolonies on both substrata with individual cells translocating between the colonies. The length of time bacteria were exposed to and their density at a surface influenced behavioral traits, with diverse and distinctive species-specific behavioral events.
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  • 12
    ISSN: 1573-0778
    Keywords: continuous culture ; death ; hybridoma ; lactate dehydrogenase ; lysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The death of the hybridoma VO 208 in a continuous culture at pH 7 and 6.8 was investigated by measuring both the appearance of visible dead cells which do not exclude the trypan blue dye and the release of lactate dehydrogenase (LDH) in the culture medium. The intracellular LDH was found to be completely released either when live cells lysed or when they were transformed into visible dead cells. No significant lysis of blue dead cells could be observed at the two different pH. Using a LDH balance over the culture system, cell lysis was found negligible at pH 7, but accounted for 20% of the total cell death at pH 6.8. A methodology is proposed to evaluate the rate constants of hybridoma lysis and total death. For the investigated cell line in continuous culture, the calculated total cell death rate constant was found to increase from 0.002 h−1 to 0.01 h−1 when decreasing the pH from 7 to 6.8.
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  • 13
    ISSN: 1573-0778
    Keywords: antibody production ; continuous culture ; culture age ; hybridoma cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The stability of the hybridoma cell line MN12 in a long-term homogeneous continuous culture was studied using a panel of analytical methods. These include two flow cytometry methods, for the determination of relative cytoplasmic and membrane IgG content. In addition, the antibody production was determined by an ELISA, and the metabolic state of the cells was determined by means of glucose consumption and lactate production. These results indicate a possible selection of variants of MN12 hybridoma cells with an overall aerobic metabolism, but with a higher glucose consumption rate and a higher lactate production rate. These variants are mainly characterized by a different membrane IgG content and cytoplasmic antibody content. These changes may possibly be affected by the culture age.
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  • 14
    ISSN: 1573-0778
    Keywords: animal cell culture ; hybridoma ; monoclonal antibody ; packed bed reactor ; continuous culture ; perfusion ; protein-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The present work demonstrates the utility of packed bed reactors for the production of monoclonal antibody. We present data from a continuous process run for the production of over 100 grams of antibody, using serum-free medium. An additional pilot run also demonstrates the potential for continued antibody production under protein-free conditions, using a standard basal medium.
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  • 15
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    Springer
    Cytotechnology 5 (1991), S. 201-209 
    ISSN: 1573-0778
    Keywords: continuous culture ; monoclonal antibodies ; passive MCAB release
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Evidence is presented for the passive release of monoclonal antibodies (MCAB) from hybridoma cells grown in either batch or continuous-flow culture. This release is promoted at room temperature. Passively released MCAB is indistinguishable from that released by actively growing cells, as judged by SDS-polyacrylamide gel electrophoresis. The significance of these observations in relation to the continuous culture of hybridoma cells is discussed. Maximum MCAB content of TB/C3 hybridoma cells is about 55pg per cell, any additional MCAB produced is secreted.
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  • 16
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    Springer
    Cytotechnology 7 (1991), S. 103-112 
    ISSN: 1573-0778
    Keywords: adaptation ; ammonia ; hybridoma ; continuous culture ; serum-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Using two mouse-mouse hybridoma cell lines, the response to ammonia step and serial changes was investigated in batch and continuous cultures with serum-free medium. The inhibitory effect of ammonia on cell growth depended on the cultivation mode, and differed markedly between cell lines. The cell line, 4C10B6 producing IgG monoclonal antibody against Pseudomonas, showed a high adaptation ability to ammonia. The 4C10B6 cells could grow under ammonia concentration as high as 21 mmol/l NH4Cl with a viability of 80% in the continuous culture with serial increase in ammonia concentration. Whereas, in the batch culture with ammonia step change the cell growth completely ceased at 12 mmol/l NH4Cl. The other cell line, TO-405 producing IgG monoclonal antibody against hepatitis B surface antigen, could not adapt to ammonia, and the cell growth did not occur at 9 mmol/l NH4Cl even under the ammonia serial change.
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  • 17
    ISSN: 1573-0778
    Keywords: cell cycle ; CHO ; continuous culture ; flow cytometry ; perfusion culture ; spin-filter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The importance of cell cycle analysis in cell culture development has been widely recognised. Whether such analysis is useful in indicating future performance of high cell density culture is uncertain. Using flow cytometric approach to address this question, we utilised the fraction of cells in the S phase to control specific growth rate and productivity in spin filter perfusion cultures and found a significant increase in the accumulated interferon-γ over that obtained from the nutrient-based controlled fed culture. While a general decrease with time exists in both percentage of S phase cells and specific growth rate, a clear oscillatory behaviour of both parameters is found in perfusion cultures.
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  • 18
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    Springer
    Cytotechnology 29 (1999), S. 71-84 
    ISSN: 1573-0778
    Keywords: bioreactor ; continuous culture ; hybridoma cells ; hyperosmolality ; monoclonal antibody production ; non-producing subpopulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In this paper, we propose an alternative strategy to the ones proposed before (Oh et al., 1993; Øyaas et al., 1994a) to get real increases of global final antibody titer and production at hyperosmotic stress, by reducing the detrimental effect of such a stress on cell growth, and conserving the stimulating effect on antibody production. It consists of cultivating the cells in continuous culture and increasing the osmolality stepwise. In this way, the cells could progressively adapt to the higher osmolality at each step and antibody titers could be nearly doubled at 370 and 400 mOsm kg-1, compared to the standard osmolality of 335 mOsm kg-1. Surprisingly, the stimulation of antibody production was not confirmed for higher osmolalities, 425 and 450 mOsm kg- 1, despite the minor negative effect on cell growth. Intracellular IgG analysis by flow cytometry revealed at these osmolalities a significant population of non-producing cells. However, even when taking into account this non-producing population, a stimulating effect on antibody production could not be shown at these highest osmolalities. It seems to us that osmolality has a significant effect on the appearance of these non-producing cells, since they were not observed in continuous cultures at standard osmolality, of comparable duration and at an even higher dilution rate. The appearance of the non-producing cells coincides furthermore with modifications of the synthesised antibody, as shown by electrophoretic techniques. It is however not really clear if these two observations reflect actually the same phenomenon. Hyperosmolality affects the cell behaviour in continuous culture in multiple ways, independently of the growth rate, counting all at least partially for the observed stimulation of antibody production: acceleration of the amino acid, and in particular the glutamine metabolism, increase of the cell volume, increase of the intracellular pH and accumulation of cells in the G1 cell cycle phase.
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  • 19
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    Cytotechnology 31 (1999), S. 243-254 
    ISSN: 1573-0778
    Keywords: continuous culture ; growth inhibition ; osmolality ; perfusion culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Continuous culture is frequently used in the cultivation of mammalian cells for the manufacturing of recombinant protein pharmaceuticals. In such operations a large volume of medium is turned over each day, especially in the case where cell recycle, or perfusion cultivation, is practiced. In principle, the volumetric throughput of medium can be reduced by using a more concentrated feed while maintaining the same nutrient provision rate. Overall, the medium components are divided into two categories: ‘consumable nutrients' and ‘unconsumable inorganic bulk salts’. In such fortified medium, the concentrations of consumable nutrients, but not bulk salts, are increased. With a stoichiometrically-balanced medium, the large amount of nutrients fed into the culture is largely consumed by cells to give rise to residual concentrations of these nutrients in their optimal range. However, unless care is taken to initiate the continuous culture, overshoot of nutrients may occur during the transient period. The high nutrient concentration during overshoot may be inhibitory by itself, or the resulting high osmolality may retard the growth. Using a mathematical model that incorporates the growth inhibitory effect of high osmolality we demonstrate such a potentially catastrophic effect of nutrient and osmolality overshoot by simulation. To avoid overshoot a controlled nutrient feeding scheme should be devised at the initiation of continuous culture.
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  • 20
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    Journal of industrial microbiology and biotechnology 15 (1995), S. 257-262 
    ISSN: 1476-5535
    Keywords: bacteria ; interaction ; biofilm ; mixed-species ; community
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Interactions among bacterial populations can have a profound influence on the structure and physiology of microbial communities. Interspecies microbial interactions begin to influence a biofilm during the initial stages of formation, bacterial attachment and surface colonization, and continue to influence the structure and physiology of the biofilm as it develops. Although the majority of research on bacterial interactions has utilized planktonic communities, the characteristics of biofilm growth (cell positions that are relatively stable and local areas of hindered diffusion) suggest that interspecies interactions may be more significant in biofilms.
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  • 21
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    Journal of industrial microbiology and biotechnology 15 (1995), S. 339-346 
    ISSN: 1476-5535
    Keywords: polysaccharides ; bacterial capsule ; biofilm
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract There has been much written on bacterial exopolysaccharides (EPS) and their role in virulence. Less has been published regarding EPS in free living species. This review focuses on that subject, emphasizing their functions in the environment and the use of antibody probes to study them.
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  • 22
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    Journal of industrial microbiology and biotechnology 16 (1996), S. 48-56 
    ISSN: 1476-5535
    Keywords: Staphylococcus epidermidis ; biofilm ; laser scanning confocal microscopy ; slime ; lectin marker
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A new dual fluorescence technique is described which, when combined with scanning confocal laser microscopy (SCLM), can be used to visualize the components of biofilm produced byStaphylococcus epidermidis. Chemostat cultures of RP62A (a well-characterized slime-producing strain ofS. epidermidis) were used to produce mature biofilm on polyvinylcholoride (PVC) disks immobilized in a modified Robbins device using a ‘seed’ and ‘feed’ model system. Serial horizontal and vertical optical thin sections, as well as three-dimensional computer reconstructions, were obtained onin situ biofilm using the dual fluorescence procedure. Bacteria were visualized by green autofluorescence excited at 488 nm with an Argon laser. Cell-associated and exocellular matrix material (slime) was visualized by red fluorescence excited at 568 nm with a Krypton laser after interaction of the biofilm with Texas Red-labeled wheat germ agglutinin which is a slime-specific lectin marker. Structural analysis revealed that the cocci grew in slime-embedded cell clusters forming distinct conical-shaped microcolonies. Interspersed open channels served to connect the bulk liquid with the deepest layers of the mature, hydrated biofilm which increased overall surface area and likely facilitated the exchange of nutrients and waste products throughout the biofilm. The combined dual fluorescence technique and SCLM is potentially useful as a specific noninvasive tool for studying the effect of antimicrobial agents on the process of biofilm formation and for the characterization of the architecture ofS. epidermidis biofilm formedin vivo andin vitro on medical grade virgin or modified inert polymer surfaces.
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  • 23
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    Journal of industrial microbiology and biotechnology 17 (1996), S. 6-10 
    ISSN: 1476-5535
    Keywords: biofilm ; cooling water ; microbiologically influenced corrosion ; microbial fouling ; stainless steel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Coupons of stainless steel type AISI-304 were exposed to the industrial cooling system of a petrochemical plant fed by seawater from the Guanabara Bay, Rio de Janeiro, Brazil, in order to study thein situ formation of biofilms. Bacteria, microalgae and fungi were detected on the coupons as soon as 48 h after exposure. Their respective numbers were determined at times 48, 96 and 192 h and over the following 8 weeks. Aerobic, anaerobic and sulfate-reducing bacteria were quantified according to the technique of the most probable number, and fungi by the pour plate technique. The number of microorganisms present in the forming biofilm varied over the experimental period, reaching maximal levels of 14×1011 cells cm−2, 30×1013 cells cm−2, 38×1011 cells cm−2 and 63×105 cells cm−2, respectively, for aerobic bacteria, anaerobic bacteria, sulfate-reducing bacteria and fungi, and the dynamics of this variation depended on the group of microorganisms.Bacillus sp,Escherichia coli, Serratia sp andPseudomonas putrefaciens were identified among the aerobic bacteria isolated. Additionally, microalgae and bacteria of the genusGallionella were also detected. Nonetheless, no evidence of corrosion was found on the stainless steel type AISI-304 coupons over the experimental period.
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  • 24
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    Journal of industrial microbiology and biotechnology 15 (1995), S. 169-175 
    ISSN: 1476-5535
    Keywords: dental plaque ; biofilm ; adhesion ; co-aggregation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Dental plaque is the diverse microbial community found on the tooth surface embedded in a matrix of polymers of bacterial and salivary origin. Once a tooth surface is cleaned, a conditioning film of proteins and glycoproteins is adsorbed rapidly to the tooth surface. Plaque formation involves the interaction between early bacterial colonisers and this film (the acquired enamel pellicle). To facilitate colonisation of the tooth surface, some receptors on salivary molecules are only exposed to bacteria once the molecule is adsorbed to a surface. Subsequently, secondary colonisers adhere to the already attached early colonisers (co-aggregation) through specific molecular interactions. These can involve protein-protein or carbohydrate-protein (lectin) interactions, and this process contributes to determining the pattern of bacterial succession. As the biofilm develops, gradients in biologically significant factors develop, and these permit the co-existence of species that would be incompatible with each other in a homogeneous environment. Dental plaque develops naturally, but it is also associated with two of the most prevalent diseases affecting industrialised societies (caries and periodontal diseases). Future strategies to control dental plaque will be targeted to interfering with the formation, structure and pattern of development of this biofilm.
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  • 25
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    Journal of industrial microbiology and biotechnology 15 (1995), S. 263-276 
    ISSN: 1476-5535
    Keywords: biofilm ; on-line monitoring ; nondestructive monitoring ; microscopy ; Fourier-transform infrared spectrometry ; bioluminescence ; microelectrode ; quartz crystal microbalance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A fundamental requirement for the understanding and control of biofilms is the continuous nondestructive monitoring of biofilm processes. This paper reviews research analytical techniques that monitor biofilm processes in a continuous nondestructive manner and that could also be modified for industrial applications. To be considered ‘continuous’ and ‘nondestructive’ for the purpose of this review a technique must: (a) function in an aqueous system; (b) not require sample removal; (c) minimize signal from organisms or contaminants in the bulk phase; and (d) provide real-time data. Various microscopic, spectrochemical, electrochemical, and piezoelectrical analysis methods fulfill these criteria. These techniques monitor the formation of biofilms, the physiology of the microorganisms within biofilms, and/or the interaction of the biofilms with their environment. It is hoped that this review will stimulate development and use of biofilm monitoring techniques in industrial and environmental settings.
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  • 26
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    Journal of industrial microbiology and biotechnology 15 (1995), S. 347-351 
    ISSN: 1476-5535
    Keywords: polysaccharides ; bacterial capsule ; adhesion ; biofilm
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Hyphomonas MHS-3 is a biphasic, marine bacterium that synthesizes an exopolysaccharide (EPS) capsule, which has a role in attaching the adherent, prosthecate developmental stages to solid substrata. To correlate structure with function, we characterized this integral EPS. It has a relatively homogeneous molecular weight of approximately 60000 daltons, is acidic, and putatively contains large concentrations ofN-acetylgalactosamine (GalNAc). The theoretical identity of the anionic component of the polymer, and the similarities betweenHyphomonas MHS-3 EPS and other adhesive marine/aquatic bacterial EPS are discussed.
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  • 27
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    Journal of industrial microbiology and biotechnology 15 (1995), S. 391-396 
    ISSN: 1476-5535
    Keywords: biofilm ; prevention ; polymer modification ; glow discharge treatment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Bacterial biofilm formation on synthetic polymers plays an important role in industry and in modern medicine, leading, for example, to difficult-to-treat infections caused by colonized foreign bodies. Prevention of biofilm formation is a necessary step in the successful prophylaxis of such infections. One approach is to inhibit bacterial adherence by polymer surface modification. We have investigated polymer modification by glow discharge treatment in order to study the influence of the modified surface on bacterial adherence. Surface roughness, surface charge density and contact angles of the modified polymers were determined and related to the adherence ofStaphylococcus epidermidis KH6. Although no influence of surface roughness and charge density on bacterial adherence was noticed, a correlation between the free enthalpy of adhesion (estimated from contact angle measurements) and adherence was observed. There seems to exist a certain minimum bacterial adherence, independent of the nature of the polymer surface. Modified polymers with negative surface charge allow for bacterial adherence close to the adherence minimum. These polymers could be improved further by the ionic bonding of silver ions to the surface. Such antimicrobial polymers are able to prevent bacterial colonization, which is a prerequisite for biofilm formation. It is suggested that modification of polymers and subsequent surface coupling of antimicrobials might be an effective approach for the prevention of bacterial biofilm formation.
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  • 28
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    Journal of industrial microbiology and biotechnology 16 (1996), S. 241-248 
    ISSN: 1476-5535
    Keywords: ethanol ; biofilm ; Zymomonas ; Saccharomyces ; Streptomyces ; plastic composite-supports
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Biofilms are a natural form of cell immobilization that result from microbial attachment to solid supports. Biofilm reactors with polypropylene composite-supports containing up to 25% (w/w) of various agricultural materials (corn hulls, cellulose, oat hulls, soybean hulls or starch) and nutrients (soybean flour or zein) were used for ethanol production. Pure cultures ofZymomonas mobilis, ATCC 31821 orSaccharomyces cerevisiae ATCC 24859 and mixed cultures with either of these ethanol-producing microorganisms and the biofilm-formingStreptomyces viridosporus T7A ATCC 39115 were evaluated. An ethanol productivity of 374g L−1 h−1 (44% yield) was obtained on polypropylene composite-supports of soybean hull-zein-polypropylene by usingZ. mobilis, whereas mixed-culture fermentations withS. viridosporus resulted in ethanol productivity of 147.5 g L−1 h−1 when polypropylene composite-supports of corn starch-soybean flour were used. WithS. cerevisiae, maximum productivity of 40 g L−1 h−1 (47% yield) was obtained on polypropylene composite-supports of soybean hull-soybean flour, whereas mixed-culture fermentation withS. viridosporus resulted in ethanol productivity of 190g L−1 h−1 (35% yield) when polypropylene composite-supports of oat hull-polypropylene were used. The maximum productivities obtained without supports (suspension culture) were 124 g L−1 h−1 and 5 g L−1 h−1 withZ. mobilis andS. cerevisiae, respectively. Therefore, forZ. mobilis andS. cerevisiae, ethanol productivities in biofilm fermentations were three- and eight-fold higher than suspension culture fermentations, respectively. Biofilm formation on the chips was detected by weight change and Gram staining of the support material at the end of the fermentation. The ethanol production rate and concentrations were consistently greater in biofilm reactors than in suspension cultures.
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  • 29
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    World journal of microbiology and biotechnology 14 (1997), S. 113-118 
    ISSN: 1573-0972
    Keywords: Aspergillus ; continuous culture ; glucoamylase ; growth ; fungi ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Maltose-limited continuous culture of Aspergillus niger was carried out with potassium nitrate to investigate growth and glucoamylase formation characteristics. Glucoamylase production was dependent on the specific growth rate. The maximal amount of glucoamylase (units/l and U/g dry weight) was obtained at μ=0.08h−1, and the maximum specific rate of production (units/g/dry weight per hour) was at μ=0.2h−1. The maintenance coefficients (ms and mATP) were higher than for some other fungi. Maximal growth yields on substrate, oxygen and ATP (Yxsm, YxO2m and Yxam) were very efficient (high) and the value of Yxam, which cannot exceed the theoretical maximal value, is obtained when a P/O ratio of 1:1 is assumed. This indicates that biomass formation is energetically inexpensive and most of the expended energy has to be invested in the process of glucoamylase excretion.
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  • 30
    ISSN: 1573-0972
    Keywords: Acetic acid production ; carbon metabolism ; continuous culture ; Escherichia coli ; metabolic engineering
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The growth kinetics of an Escherichia coli wild type strain and two derivative mutants were examined in batch cultures and in glucose-limited chemostats. One mutant (PB12) had an inactive phosphotranferase transport system and the other (PB25) had interrupted pykA and pykF genes that code for the two pyruvate kinase isoenzymes. In both batch and continuous culture, important differences in acetic acid accumulation and other metabolic activities were found. Compared to the wild type strain, we observed a reduction in acetic acid accumulation of 25 and 80% in PB25 and PB12 strains respectively, in batch culture. Continuous culture experiments revealed that compared to the other two strains, PB25 accumulated less acetic acid as a function of dilution rate. In continuous cultures, oxidoreductase metabolic activities were substantially affected in the two mutant strains. These changes in turn were reflected in different levels of biomass and CO2 production, and in oxygen consumption.
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  • 31
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    Biotechnology and Bioengineering 59 (1998), S. 261-272 
    ISSN: 0006-3592
    Keywords: effective diffusive permeability ; diffusion coefficient ; biofilm ; cell density ; review ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Experimental measurements of effective diffusive permeabilities and effective diffusion coefficients in biofilms are reviewed. Effective diffusive permeabilities, the parameter appropriate to the analysis of reaction-diffusion interactions, depend on solute type and biofilm density. Three categories of solute physical chemistry with distinct diffusive properties were distinguished by the present analysis. In order of descending mean relative effective diffusive permeability (De/Daq) these were inorganic anions or cations (0.56), nonpolar solutes with molecular weights of 44 or less (0.43), and organic solutes of molecular weight greater than 44 (0.29). Effective diffusive permeabilities decrease sharply with increasing biomass volume fraction suggesting a serial resistance model of diffusion in biofilms as proposed by Hinson and Kocher (1996). A conceptual model of biofilm structure is proposed in which each cell is surrounded by a restricted permeability envelope. Effective diffusion coefficients, which are appropriate to the analysis of transient penetration of nonreactive solutes, are generally similar to effective diffusive permeabilities in biofilms of similar composition. In three studies that examine diffusion of very large molecular weight solutes ( 〉 5000) in biofilms, the average ratio of the relative effective diffusion coefficient of the large solute to the relative effective diffusion coefficient of either sucrose or fluorescein was 0.64, 0.61, and 0.36. It is proposed that large solutes are effectively excluded from microbial cells, that small solutes partition into and diffuse within cells, and that ionic solutes are excluded from cells but exhibit increased diffusive permeability (but decreased effective diffusion coefficients) due to sorption to the biofilm matrix. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:261-272, 1998.
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  • 32
    ISSN: 1573-0778
    Keywords: continuous culture ; free solution capillary electrophoresis ; hollow fibers ; monoclonal antibody characterization ; protein-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The performance of a protein-free medium was compared in culture flasks with a serum-supplemented medium and with a serum free medium in terms of cell growth and monoclonal antibody production by a murine hybridoma. We present results of continuous production in hollow fiber culture systems using serum-free medium and protein-free medium. In protein-free medium, it has been possible to produce large quantities of monoclonal antibody with a productivity similar to that obtained in serum-free medium. After a two steps purification process, monoclonal antibodies were characterized by SDS-PAGE, High Performance Size Exclusion Chromatography and Free Solution Capillary Electrophoresis. SDS-PAGE and high performance chromatography analysis have showed that purified monoclonal antibodies produced in serum-free medium or protein-free medium were similar. Furthermore, Capillary Electrophoresis characterization revealed that both MAbs were constituted by three isoforms with equivalent electrophoretic mobilities.
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  • 33
    ISSN: 1573-0778
    Keywords: antibody production ; carbon dioxide ; cell metabolism ; continuous culture ; inhibition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract CO2 partial pressure (pCO2) in industrial cell culture reactors may reach 150–200 mm Hg, which can significantly inhibit cell growth and recombinant protein production. Due to equilibrium with bicarbonate, increased pCO2 at constant pH results in a proportional increase in osmolality. Hybridoma AB2-143.2 cell growth rate decreased with increasing pCO2 in well-plate culture, with a 45% decrease at 195 mm Hg with partial osmolality compensation (to 361 mOsm kg- 1). Inhibition was more extensive without osmolality compensation, with a 63% decrease in growth rate at 195 mm Hg and 415 mOsm kg-1. Also, the hybridoma death rate increased with increasing pCO2, with 31- and 64-fold increases at 250 mm Hg pCO2 for 401 and 469 mOsm kg- 1, respectively. The specific glucose consumption and lactate production rates were 40–50% lower at 140 mm Hg pCO2. However, there was little further inhibition of glycolysis at higher pCO2. The specific antibody production rate was not significantly affected by pCO2 or osmolality within the range tested. Hybridomas were also exposed to elevated pCO2 in continuous culture. The viable cell density decreased by 25–40% at 140 mm Hg. In contrast to the well-plate cultures, the death rate was lower at the new steady state at 140 mm Hg. This was probably due to higher residual nutrient and lower byproduct levels at the lower cell density (at the same dilution rate), and was associated with increased cell-specific glucose and oxygen uptake. Thus, the apparent effects of pCO2 may vary with the culture system.
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  • 34
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    Cytotechnology 10 (1992), S. 147-155 
    ISSN: 1573-0778
    Keywords: batch culture ; continuous culture ; hybridoma ; kinetics ; specific rates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract For the mouse hybridoma cell line VO 208, kinetics of growth, consumption of glucose and glutamine, and production of lactate, ammonia and antibodies were compared in batch and continuous cultures. At a given specific growth rate, different metabolic activities were observed: a 40% lower glucose and glutamine consumption rate, but a 70% higher antibody production rate in continuous than in batch culture. Much higher metabolic rates were also measured during the initial lag phase of the batch culture. When representing the variation of the specific antibody production rate as a function of the specific growth rate, there was a positive association between growth and antibody production in the batch culture, but a negative association during the transient phase of the continuous culture. The kinetic differences between cellular metabolism in batch and continuous cultures may be result of modifications in the physiology and metabolism of cells which, in continuous cultures, were extensively exposed to glucose limitations.
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  • 35
    ISSN: 1573-0778
    Keywords: antithrombin III ; mammalian cell culture ; continuous culture ; capillary electrophoresis ; product monitoring ; recombinant protein ; temperature influence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The importance of mammalian cell cultures for biotechnological production processes is steadily increasing, despite the high demands of these organisms on their culture conditions. Efforts towards a more efficient bioprocess generally concentrate on maximizing the culture's life time, the cell number, and the product concentration. Here recombinant BHK 21 c13 cells are used to produce rh-AT III, an anticoagulant of high therapeutic value. The influence of the process mode (batch, repeated batch, continuous perfusion) and the process temperature (30°C vs. 37°C) on the above mentioned parameters is investigated. It is possible to increase the length of the culture from 140 h (batch) to more than 500 h (continuous perfusion culture), while concomitantly increasing the cell density from 0.72 106/ml (batch) to 2.27 106/ml (repeated batch) and 2.87 106/ml (continuous perfusion culture). The accumulation of toxic metabolites, such as lactate, can be curtailed by reducing the bioreactor temperature from 37°C to 30°C during the later part of the exponential growth phase. Fast and reliable product monitoring became essential during process optimization. Capillary zone electrophoresis (CZE) in uncoated fused silica capillaries was studied for that purpose and compared to the standard ELISA. Under optimized conditions an AT III quantification could be done within 2 min with CZE. The detection limit was 5 μg/ml. A relative standard deviation of less than 0.9% was calculated. The detection limit could be lowered by one order of magnitude by using a two dimensional system, where an liquid chromatographic (LC) system is coupled to the CZE. Concomitantly the resolution is improved. The two-dimensional analysis required 5 min. Membrane adsorbers (MA) were used as stationary phase in the LC-system, to allow the application of high flow rates (5–10 ml/min). The correlation between the LC-CZE analysis and the standard AT III-ELISA was excellent, with r2: 0.965. Using the assay for at line product monitoring, it is shown, that the process temperature is of no consequence for the productivity whereas the process mode strongly influences this parameter.
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  • 36
    ISSN: 1573-0778
    Keywords: cell cycle ; CHO cells ; continuous culture ; SV40 promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Foreign protein expression from the commonly used SV40 promoter has been found to be primarily during the S-phase of the cell cycle. Simple mathematical models with this cell cycle phase dependent expression of foreign protein suggest that the specific production rate will be proportional to the cell growth rate, which is particularly disadvantageous in high cell density fed-batch or perfusion bioreactors. In this study we investigate this predicted relationship between the production rate and growth rate by culturing recombinant CHO cells in a continuous suspension bioreactor. One CHO cell line, GS-26, has been stably transfected with the plasmid pSVgal, which contains the E. coli lac Z gene under the control of the SV40 promoter. This GS-26 cell line was grown in suspension cultures over a range of specific growth rates in batch and continuous modes. The intracellular β-galactosidase activity was assayed using a standard spectrophotometric method after breaking the cells open and releasing the enzyme. A strong growth associated relationship is found between the intracellular β-galactosidase content and the specific growth rate in batch and continuous cultures, as predicted.
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  • 37
    ISSN: 1573-0778
    Keywords: adenovirus major late promoter ; β-galactosidase ; Chinese hamster ovary cells ; continuous culture ; G1 phase expression ; inverse-growth associated production
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Protein synthesis in mammalian cells can be observed in two strikingly different patterns: 1) production of monoclonal antibodies in hybridoma cultures is typically inverse growth associated and 2) production of most therapeutic glycoproteins in recombinant mammalian cell cultures is found to be growth associated. Production of monoclonal antibodies has been easily maximized by culturing hybridoma cells at very low growth rates in high cell density fed- batch or perfusion bioreactors. Applying the same bioreactor techniques to recombinant mammalian cell cultures results in drastically reduced production rates due to their growth associated production kinetics. Optimization of such growth associated production requires high cell growth conditions, such as in repeated batch cultures or chemostat cultures with attendant excess biomass synthesis. Our recent research has demonstrated that this growth associated production in recombinant Chinese hamster ovary (CHO) cells is related to the S (DNA synthesis)-phase specific production due to the SV40 early promoter commonly used for driving the foreign gene expression. Using the stably transfected CHO cell lines synthesizing an intracellular reporter protein under the control of SV40 early promoter, we have recently demonstrated in batch and continuous cultures that the product synthesis is growth associated. We have now replaced this S-phase specific promoter in new expression vectors with the adenovirus major late promoter which was found to be active primarily in the G1-phase and is expected to yield the desirable inverse growth associated production behavior. Our results in repeated batch cultures show that the protein synthesis kinetics in this resulting CHO cell line is indeed inverse growth associated. Results from continuous and high cell density perfusion culture experiments also indicate a strong inverse growth associated protein synthesis. The bioreactor optimization with this desirable inverse growth associated production behavior would be much simpler than bioreactor operation for cells with growth associated production.
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  • 38
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    World journal of microbiology and biotechnology 11 (1995), S. 643-645 
    ISSN: 1573-0972
    Keywords: Chemostat ; 4-chlorosalicylic acid ; continuous culture ; degradation ; microbial consortium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A mixed, stable microbial community, obtained by continuous enrichment of a sediment core using 4-chlorosalicylic acid as sole source of carbon and energy, contained 10 different bacterial species, including Klebsiella pneumonia, Pseudomonas fluorescens, P. mendocina and P. cichorii. The members of the community were grown separately on various chlorinated compounds which were readily degraded.
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  • 39
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    World journal of microbiology and biotechnology 12 (1996), S. 395-397 
    ISSN: 1573-0972
    Keywords: Biocide ; biofilm ; Hormoconis ; immunofluorescence ; Kathon FP ; stainless steel ; sulphate-reducing bacteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Biofilms containing single or mixed cultures of the fungus Hormoconis resinae and anaerobic sulphate-reducing bacteria (SRB) on stainless steel were incubated with an isothiazolone biocide (Kathon FP) at 28°C for 24 h. H. resinae within the biofilm was enumerated by immunofluorescence microscopy using specific antiserum, and SRB were assayed by culture. Fungal numbers in mixed biofilms were considerably reduced in comparison with those in pure biofilms. The biocide was shown to be effective against H. resinae in pure biofilms at 50 and 100 ppm, but in mixed biofilms only at the higher concentration. This concentration also reduced the sessile SRB numbers by 99%.
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  • 40
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    World journal of microbiology and biotechnology 15 (1999), S. 115-117 
    ISSN: 1573-0972
    Keywords: Candida utilis ; pineapple cannery effluent ; continuous culture ; steady-state
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Candida utilis was grown on a pineapple cannery effluent in a chemostat at dilution rates ranging between 0.05 and 0.65 h−1 to establish optimal conditions for biomass production and chemical oxygen demand (COD) reduction. Sucrose, fructose and glucose were the main sugars in the effluent. Maximum value for cell yield coefficient and productivity were (0.686, gx/gs) and (2.96, gx/l/h) at a dilution rate of 0.425 and 0.475 h−1, respectively, while maximum COD reduction (98%) was attained at a dilution rate of 0.1 h−1. The maintenance coefficient attained a value of (0.093, gs/gx/h). An increase in dilution rate produced a higher protein content of the biomass.
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  • 41
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    World journal of microbiology and biotechnology 15 (1999), S. 235-238 
    ISSN: 1573-0972
    Keywords: Arthrospira platensis ; bioenergetic yield ; continuous culture ; irradiance ; specific growth rate ; mixotrophy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Reassessement of bioenergetic growth yield of Arthrospira platensis was performed by using continuous culture under both autotrophic and mixotrophic conditions. Continuous culture was carried out at dilution rates of 0.017, 0.023 and 0.030 h−1. Under these dilution rates bioenergetic yields ranged between 4.45–6.03 × 10−3 g biomass kJ−1 and between 5.42–7.46 × 10−3 g biomass kJ−1, under autotrophic and mixotrophic conditions respectively. A maximum bioenergetic yield of 8.1 × 10−3 g biomass kJ−1 using an autotrophic culture can be calculated. Pigment accumulation (chlorophyll a and carotenoids) may be related to light irradiance, reaching a maximum pigment concentration under light saturation irradiance. Phycocyanin concentration increased during light limitation.
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  • 42
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    Biotechnology and Bioengineering 38 (1991), S. 877-882 
    ISSN: 0006-3592
    Keywords: microtechnique ; microprobe ; biofilm ; dissolved oxygen concentration ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel in situ microtechnique allows evaluating parameters of diffusion-controlled reactions in biofilms. A microprobe, 15 μm in diameter, was used to simultaneously measure the dissolved oxygen concentration and the optical density at different depths in a submerged biofilm. Based on the results, the biofilm diffusion coefficient for dissolved oxygen, Df the dissolved oxygen flux through the biofilm surface, J02, and the half velocity coefficient, Ks, have been calculated.
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  • 43
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    Biotechnology and Bioengineering 38 (1991), S. 781-787 
    ISSN: 0006-3592
    Keywords: hybridoma culture ; monoclonal antibody production ; perfusion culture ; continuous culture ; cell cycle ; tangential filtration ; cell separation ; nuclepore membranes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In order to elucidate the hybridoma life cycle and the limiting factors in perfusion systems, we performed cultures in a stirred tank bioreactor, coupled to an external tangential flow filtration unit. Cell density and antibody production in perfusion were consistent with previous studies. The average life span of the cells (2.1-2.2 days), antibody, productivity per cell produced (30-38 mg/109 cells) and cell size diameter evolution appeared similar to values observed in batch cultures. These observations highly suggest a similar “grow or die” life cycle. Cell and antibody production, strictly related to the medium perfusion rate, seem to be under the control of the nutrient availability. A hypothesis to explain such a life cycle of hybridoma cells in perfusion systems and a model for viable and dead cell density is proposed.
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  • 44
    ISSN: 0006-3592
    Keywords: methylotroph ; continuous culture ; oxiturbidostat ; growth-limiting substrate pulse ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The growth characteristics of a chemostat culture of the obligate methylotrophic bacterium Methylobacillus flagellatum have been determined. Steady-state cultures growing at a rate of 0.73-0.74 h-1, equal to the maximal growth rate, were obtained under oxyturbidostat cultivation conditions. The response of a chemostat culture to a pulse increase of methanol concentration was studied. It was shown that slow and rapidly growing cultures of M. flagellatum responded differently to pulse methanol addition. The growth characteristics of slow-growing cultures decreased after methanol addition compared to those of stationary chemostat cultures. The growth characteristics of rapidly growing cultures were practically unchanged with and without pulse methanol addition.
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  • 45
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    Biotechnology and Bioengineering 41 (1993), S. 43-54 
    ISSN: 0006-3592
    Keywords: acetic acid ; alkaline protease ; Bacilus firmus ; continuous culture ; extracellular enzymes ; carbon/nitrogen/phosphorus limitation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Invariance of culture conditions in steady state continuous cultures make these a very valuable tool to study the influence of various culture parameters on cell growth and synthesis of primary and secondary metabolites. The result of a parametric study on production of protease in continuous suspension cultures of Bacillus firmus NRS 783 are reported in this article. This strain is a superior producer of an alkaline protease with major application in the detergent industry. The parameters investigated include dilution rate and concentrations of yeast extract, ammonium, and inorganic phosphate in the bioreactor feed, glucose being the principal carbon source in all experiments. The regulatory effects of the key culture parameters on cell growth, synthesis and secretion of protease, and production of acetic acid are investigated. The relations among the specific cell growth rate, specific utilization rates of the principal carbon, nitrogen, and phosphorous sources, and specific production rates of two nonbiomass products, viz., acetic acid and protease, are examined, and the effects of the manipulated culture parameters on these relations, specific protease activity, and yields of cell mass, protease, and acetic acid on the basis of the principal carbon, nitrogen, and phosphorous sources are studied. An increase in dilution rate led to increases in specific utilization rates of the principal carbon, nitrogen, and phosphorous sources and specific production rates of acetic acid and protease and decreases in bulk activities/concentrations of the three products (acetic acid, cell mass, and protease). As a result, the productivities of the three species were maximized at an intermediate dilution rate. Increased supply of yeast extract (a rich source of amino acids, proteins, and vitamins, besides being an additional source of carbon, nitrogen, and phosphorus) promoted cell mass formation but reduced protease production per unit cell mass. Increased supply of nitrogen and phosphorous sources stimulated protease synthesis up to certain threshold levels and repressed the enzyme synthesis beyond the threshold levels. With increased supply of the nitrogen source, the phosphorous source was more efficiently utilized for cell growth and protease synthesis. Stable maintenance of continuous cultures of B. firmus over prolonged period is demonstrated in this study. © 1993 John Wiley & Sons, Inc.
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  • 46
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    Biotechnology and Bioengineering 41 (1993), S. 572-580 
    ISSN: 0006-3592
    Keywords: phenol degradation ; continuous culture ; Pseudomonas putida ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pure cultures of Pseudomonas putida (ATCC 17484) were grown in continuous culture on phenol at dilution rates of 0.074-0.085 h-1 and subjected to step increases in phenol feed concentration. Three distinct patterns of dynamic response were obtained depending on the size of the step change used: low level, moderate level, or high level. During low level responses no accumulations of phenol or non-phenol, non-glucose-dissolved organic carbon, DOC(NGP), were observed. Moderate level responses were characterized by the transient accumulation of DOC(NGP) with a significant delay prior to phenol leakage. High level responses demonstrated a rapid onset of phenol leakage and no apparent accumulations of DOC(NGP). The addition of phenol to a continuous culture of the same organism on glucose did not result in transient DOC(NGP) accumulations, although transient phenol levels exceeded 90 mg l-1. These results were consistent with intermediate metabolite production during phenol step tests coupled with substrate-inhibited phenol uptake and suggested that traditional kinetic models based on the Haldane equation may be inadequate for describing the dynamics of phenol degrading systems. © 1993 John Wiley & Sons, Inc.
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  • 47
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    Biotechnology and Bioengineering 43 (1994), S. 434-438 
    ISSN: 0006-3592
    Keywords: hybridoma ; continuous culture ; ammonia ; growth inhibition ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The nature and temporal development of ammonia inhbition were investigated in batch, fed-batch, and continuous cultures. Significant inhibition was observed when cells were inoculated in serum-containing or chemically defined medium containing more than 2 mM of ammonia. In contrast, no inhibition was observed at greater than 10 mM when the ammonia concentration was gradually increased over the span of a batch culture by feeding ammonium chloride. Strong growth inhibition was observed after each of five step changes (2.8 → 3.7 → 4.0 → 4.9 → 7.7 → 13.5 mM) in continuous culture. Following a period of adaptation at each higher value, the viable cell density stabilized at a new lower value. The lowering in viable cell density was caused by an increase in specific death rate and a decreased cell yield on glucose, glutamine, and oxygen. Increased ammonia concentration had little or no effect on the steady-state specific growth kinetics or specific antibody productivity. © 1994 John Wiley & Sons, Inc.
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  • 48
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    Biotechnology and Bioengineering 41 (1993), S. 728-735 
    ISSN: 0006-3592
    Keywords: biofilm ; shear stress ; substrate loading ; biofilm detachment ; Pseudomonas aeruginosa ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: One of the least understood processes affecting biofilm accumulation is detachment. Detachment is the removal of cells and cell products from an established biofilm and subsequent entrainment in the bulk liquid. The goal of this research was to determine the effects of shear stress and substrate loading rate on the rate of biofilm detachment.Monopopulation Pseudomonas aeruginosa and undefined mixed population biofilms were grown on glucose in a RotoTorque biofilm reactor. Three levels of shear stress and substrate loading rate were used to determine their effects on the rate of detachment. Suspended cell concentrations were monitored to determine detachment rates, while other variables were measured to determine their influence on the detachment rate. Results indicate that detachment rate is directly related to biofilm growth rate and that factors which limit growth rate will also limit detachment rate. No significant influence of shear on detachment rate was observed.A new kinetic expression that incorporates substrate utilization rate, yield, and biofilm thickness was compared to published detachment expressions and gives a better correlation of data obtained both in this research and from previous research projects, for both mono- and mixed-population biofilms. © John Wiley & Sons, Inc.
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  • 49
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    Biotechnology and Bioengineering 44 (1994), S. 194-204 
    ISSN: 0006-3592
    Keywords: biofilm ; biofilm reactors ; structure ; heterogeneity ; kinetics ; modeling ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A rotating annular reactor (Roto Torque) was used for qualitative and quantitative studied on biofilm heterogeneity. In contrast to the classic image of biofilms as smooth, homogeneous layers of biomass on a substratum, studies using various pure and mixed cultures consistently revealed more-dimensional structures that resembled dunes and ridges, among others. These heterogeneities were categorized and their underlying causes analyzed. Contrary to expectations, motility of the microorganisms not a decisive factor in determining biofilm homogeneity. Small Variations in substratum geometry homogeneity. Small variations in substratum geometry and flow patterns were clearly reflected in the biofilm pattern. Nonhomogeneous flow and shear patterns in the reactor, together with inadequate mixing resulted in significant, position-dependent differences in surface growth. It was therefore not possible to take representative samples of the attached biomass. Like many other types of reactors, the Roto Torque reactor is valuable for qualitative and morphological biofilm experiments but less suitable for quantitative physiological and kinetics studies using attached microorganisms. © 1994 John Wiley & Sons, Inc.
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  • 50
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    Biotechnology and Bioengineering 44 (1994), S. 867-879 
    ISSN: 0006-3592
    Keywords: biofilm ; microbeads ; solids retention time ; airlift reactor ; particulates ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fluorescent microparticles were used as tracer beads to measure the dynamics of solids in spherical biofilms in a biofilm airlift suspension reactor. Attachment to, release from, and penetration into the biofilms of the tracer beads were measured. The coverage of the biofilm surface was low and the steady state particle concentration on the surface was dependent on the biofilm surface characteristics. The measured attachment rate constant was identical in both experiments and appeared to be determined by the hydrodynamic conditions in the turbulent reactor. The attachment rate was much faster than the release rate of the tracer beads and, therefore, the solidsretention time in the biofilm particle is not due to a simple reversible adsorption-desorption process. The heterogeneity of the distribution oftracer beads on different sectors on the biofilm surface decreased duringthe attachment period. Due to random detachment processes the heterogeneity of the tracer bead distribution increased during the release periodThe tracer beads quickly penetrated into the biofilm and became distributed throughout the active layer of the biofilm. The observed penetration into biofilms, the nonuniform distribution on the biofilm surface, and the fast uptake and slow release of tracer beads cannot be described by a simple model based on a reversible adsorption-desorption mechanism, nor withexisting biofilm models. These biofilm models, which balance growth and advection assuming a uniform biofilm with a homogeneous surface, are inadequate for the description of the observed solids retention time in biofilms. Therefore, a new concept of biofilm dynamics is proposed, in which formation of cracks and fissures, which are rapidly filled with growing biomass, combined with nonuniform local detachment, explains the observed fast penetration into the biofilm of tracer beads, the long residence time, and the nonuniform distibution of fluorescent microparticles. © 1994 John Wiley & Sons, Inc.
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  • 51
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    Biotechnology and Bioengineering 45 (1995), S. 107-115 
    ISSN: 0006-3592
    Keywords: biofilm ; waste gas treatment ; hydrophobic microporous membrane ; mass transfer ; propene ; Xanthobacter ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel type of bioreactor for waste gas treatment has been designed. The reactor contains a microporous hydrophobic membrane to create a large interface between the waste gas and the aqueous phase. To test the new reactor, propene was chosen because of its high air/water partition coefficient, which causes a low water concentration and hampers its removal from air. Propene transfer from air to a suspension of propene-utilizing Xanthobacter Py2 cells in the membrane bioreactor proved to be controlled by mass transfer in the liquid phase. The resistance of the membrane was negligible. Simulated propene transfer rates agreed well with the experimental data. A stable biofilm of Xanthobacter Py2 developed on the membrane during prolonged operation. The propene flux into the biofilm was 1 × 10-6 mol m-2 s-1 at a propene concentration of 9.3 × 10-2 mol m-3 in the gas phase. © 1995 John Wiley & Sons, Inc.
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  • 52
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    Biotechnology and Bioengineering 44 (1994), S. 303-321 
    ISSN: 0006-3592
    Keywords: hybridoma metabolism ; continuous culture ; suspension culture ; antibody productivity ; amino acids ; vitamins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of step-change increase in the concentrations of amino acids and vitamins on the metabolism, growth, and antibody productivity of a murine hybridoma cell line grown in continuous culture on serum-free medium are presented. Additions of the amino acids cysteine with methionine, tryptophan, and isoleucine with valine and vitamin B12 (as cyanocobalamin) resulted in significant increases in viable cell concentrations. Additions of aspartate with asparagine, and threonine with vitamin B1 (as thiamine hydrochloride) resulted in significant increases in final antibody concentrations. Substantial decrease in the fraction of amino acid nitrogen excreted as ammonia occurred upon supplementation with three times the normal concentrations of branched chain amino acids. Decreases in the fraction of amino acid nitrogen converted to ammonia were paralleled by increases in the fraction converted to alanine. © 1994 John Wiley & Sons, Inc.
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  • 53
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    Biotechnology and Bioengineering 45 (1995), S. 503-510 
    ISSN: 0006-3592
    Keywords: biofilm ; thickness ; heterogeneity ; roughness ; microscopy ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The thickness variability of biofilms of Pseudomonas aeruginosa, Klebsiella pneumoniae, and the binary population combination of these two species was quantified. The experimental method involved cryoembedding biofilms with a commercial tissue embedding agent, sectioning, and applying image analysis to construct thickness profiles along linear transects (up to 1 cm in length) across the substratum. Biofilms embedded and sectioned by this method were locally as thin as a single cell attached to the surface (〈5 μm) and as thick as 1000 μm. Week-old biofilms of three different species compositions displayed distinct structural features as indicated by their mean thicknesses and by a roughness coefficient. Monopopulation biofilms of P. aeruginosa (29 μm mean thickness) or K. pneumoniae (100 μm mean thickness) were thinner than the binary population biofilm (400 μm mean thickness). A roughness coefficient developed in this investigation corroborated the qualitative visual characterization of P. aeruginosa biofilms as relatively uniformly thick (mean roughness coefficient 0.15), K. pneumoniae biofilms as patchy (mean roughness coefficient 1.14), and the binary population biofilm as intermediate (mean roughness coefficient 0.26). Whereas P. aeruginosa and binary population biofilms covered the substratum completely, significant areas of essentially bare substratum were apparent in K. pneumoniae biofilms. The patchiness of K. pneumoniae biofilms may be due to the fact that this organism is nonmotile. A spatial correlation analysis of the thickness data indicated that thickness measurements were still correlated even when separated by distances that exceeded the mean biofilm thickness. Cell aggregates, some of them hundreds of microns in size, were observed in the effluent of K. pneumoniae and binary population biofilm reactors. Measurements of thickness variability and other observations reported in this article provide a quantitative basis for analysis of microscale structural heterogeneity of biofilms. © 1995 John Wiley & Sons, Inc.
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  • 54
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    Biotechnology and Bioengineering 45 (1995), S. 524-535 
    ISSN: 0006-3592
    Keywords: respiration quotient ; carbon dioxide evolution rate ; continuous culture ; cell metabolism ; bicarbonate buffer ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The determination of the respiration quotient (RQ = CER/OUR) has not been used so far as a tool for understanding animal cell metabolism. This is due to problems in measuring the carbon dioxide evolution rate (CER) rather than the oxygen uptake rate (OUR). The determination of the CER is complicated by the use of bicarbonate in the medium. Using liquid and gas balances we have derived an equation for continuous culture to quantify the amount of CO2 that comes from the bicarbonate in the feed. Under cell-free conditions, values predicted by this equation agree within 4% with the experimental results. In continuous culture using hybridoma cells, the CO2 from the feed, as determined by an IR-gas analyzer, was found to represent a significant amount of the total measured CO2 in the off-gas (50% in a suboptimal, and 30% in high-growth medium). Furthermore, the problem of CO2 loss from the medium during medium preparation and storage was solved using both a theoretical and an experimental approach. RQ values in continuous culture were evaluated for two different growth media. Small but significant differences in RQ were measured, which were matched by differences in specific antibody rates and other metabolic quotients. In a medium with Primatone RL, an enzymatic hydrolysate of animal cell tissue that causes a more than twofold increase in cell density, the RQ was found to be 1.05, whereas in medium without Primatone RL (but containing amino acids equivalent in composition and concentration to Primatone RL) the RQ was found to be 0.97. We suggest the RQ to be a useful parameter for estimating the physiological state of cells. Its determination could be a suitable tool for both the on-line control of animal cell cultivations and the understanding of cell metabolism. © 1995 John Wiley & Sons, Inc.
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  • 55
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    Biotechnology and Bioengineering 38 (1991), S. 75-81 
    ISSN: 0006-3592
    Keywords: mycelial morphology ; Fusarium graminearum ; mycoprtein ; continuous culture ; chemostat ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The morphology of mycelial fungi in liquid culture effects culture rheology and this in turn may affect product yield. It is therefore important to understand how environmental factors influence mycelial morphology and this paper describes the effect of dilution rate on two strains of Fusarium graminearum, the relatively sparsely branched parental strain (A3/5) and a relatively highly branched “colonial” variant (C106). At any given dilution rate, the concentration of mycelial fragments present at steady state of both strains remained approximately constant with time, suggesting that mycelial fragmentation occurred in a regular manner. However, for both strains fragment concentration decreased with increasing dilution rate. The strains had a similar morphology at a dilution rate of 0.07 h-1. The length of the hyphal growth unit of A3/5 increased with increase in dilution rate, while that of C106 decreased with increase in dilution rate. At all dilution rates, C106 produced up to ten times more macroconidia than A3/5.
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  • 56
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    Biotechnology and Bioengineering 38 (1991), S. 254-259 
    ISSN: 0006-3592
    Keywords: light limitation ; shading ; maintenance ; mathematical model ; continuous culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Light-limited growth in continuous cultures of phototrophic organisms is modeled. It is assumed that light energy up-take rate depends hyperbolically on light intensity and that the maintenance costs are proportional to biomass. Modeling the light distribution caused by shading within the vessel is necessary to explain the existence of steady state in light-limited chemostats. The model fits well to experimental data from literature on light-limited chemostats and turbidostats. Attention is given to the implications of the model for the estimation of the specific maintenance rate constant in light-limited continuous cultures.
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  • 57
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    Biotechnology and Bioengineering 38 (1991), S. 557-560 
    ISSN: 0006-3592
    Keywords: levan ; continuous culture ; molecular weight ; Erwinia herbicola ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The optimal production of the fructan biopolymer levan by the bacterium Erwinia herbicola was investigated, including variations in nitrogen, carbon and phosphorous sources, pH, incubation time, culture yields up to 19% by weight produced based on conversion of sucrose as the carbon source when grown in a continuous culture system and processed by tangential flow filtration. Product identity was confirmed with gas chromatography (GC) and 13C nuclear magnetic resonance (NMR). Gel permeation chromatography (GPC) and low-angle laser light scattering (LALLS) determination of the molecular weight of the product showed a significant difference in molecular weight values dependent on the method of analysis. Analysis by GPC resulted in molecular weight one order of magnitude lower than LALLS independent of sample, underscoring the unusual nature of this biopolymer.
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  • 58
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    Biotechnology and Bioengineering 38 (1991), S. 972-976 
    ISSN: 0006-3592
    Keywords: cell culture ; antibody production ; fermentation ; continuous culture ; cell growth ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A clonal derivative of a transfectant of the SP2/O myeloma cell line producing a chimeric monoclonal antibody was maintained in steady-state, continuous culture at dilution rates ranging from 0.21 to 1.04 day-1. The steady-state values for nonviable and total cell concentrations increased as the dilution rate decreased, while the viable cell concentration was roughly independent of the dilution rate. At steady state, the specific growth rate increased and the specific death rate decreased as the dilution rate increased. The maximum specific growth rate was 1.15 day-1. Antibody production was growth associated and the specific rate of antibody production increased linearly as the specific growth rate increased.
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  • 59
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    Biotechnology and Bioengineering 38 (1991), S. 1020-1028 
    ISSN: 0006-3592
    Keywords: hybridoma ; cell culture ; continuous culture ; kinetics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A hybridoma cell line, AFP-27-P, was cultivated in continuous culture under glucose-limited conditions. The viable cell concentration, dead-cell concentration, and cell volume all varied with the dilution rate. A model previously developed for a nonproducing clone of the same cell line, AFP-27-NP, was extended to describe the behavior of the cells. The relationship between the specific growth rate and glucose concentration is described by a function similar to the Monod model. A threshold glucose concentration and a minimum specific growth rate are incorporated; the model is meaningful only at glucose concentration and a minimum specific growth rate are incorporated; the model is meaningful only at glucose concentrations and specific growth rates above these levels. The relationship between the death rate and the glucose concentration is described by an inverted Monod-type function. Furthermore, the yield coefficient based on glucose is constant in the lower range of specific growth rates and changes to a new constant value in the upper range of specific growth rates. No maintenance term for glucose consumption is used; in the plot of specific glucose consumption rate vs. specific growth rate, the line intercepts the specific growth rate at a value close to the minimum growth rate. The productivity of antibody as a function of the specific growth rate is described by a mixed type model with a noon-growth-associated term and a negative-growth-associated term. The values for the model parameters were determined from regression analysis of the steady state data.
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  • 60
    ISSN: 0006-3592
    Keywords: catabolite repression ; protein A ; membrane proteins ; continuous culture ; protein expression ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Although widely used in experimental and industrial situations, genetically engineered plasmids containing the lac promoter from Escherichia coli are subject to catabolite repression when grown in glucose-containing media. Several methods of overcoming this problem have been investigated by studying the expression of the protein A gene from Staphylococcus aureus under the control of the Escherichia coli lac promoter. When glycerol is used as a sole carbon source, the plasmid is unstable and is rapidly lost from the culture. When the bacteria are grown in chemostats under glucose limitation, the plasmid is maintained, even at high dilution rates, and the expression of protein A is similar to that observed when glycerol was used. The balance between metabolic load and protein A expression seems to be maintained by reducing the gene dose to a tolerable level. Depending on the metabolic conditions prevailing in the culture, this is achieved, either by reducing the copy number of the plasmid or in extreme cases by removing the plasmid altogether.
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  • 61
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    Biotechnology and Bioengineering 39 (1992), S. 504-510 
    ISSN: 0006-3592
    Keywords: hybridoma ; continuous culture ; dialysis ; monoclonal antibody ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Hybridoma cell growth and monoclonal antibody production in dialyzed continuous suspension culture were investigated using a 1.5-L Celligen bioreactor. Medium supplemented with 1.5% fetal bovine serum was fed directly into the reactor at a dilution rate of 0.45 d-1. Dailysis tubing with a molecular weight cut-off (MWCO) of 1000 was coiled inside the bioreactor. Fresh medium containing no serum or serum substitues passed through the dialysis tubing at flow rates of 2 to 5 L/d. The objective was to remove low molecular weight inhibitors, such as lactic acid and ammonia, by diffusion through the tubing, while continuoulsy replenishing essential nutrients by the same mechanism. Due to the low MWCO of the dialysis tubing high molecular weight components such as growth factors and antibody were not removed by the dialyzing stream. In the batch start-up phase, the monoclonal antibody (MAb) titer was almost 3 times that achieved in typical batch cultures (i.e., 170 to 180 mg/L). During dialyzed continuous operation, a substantial increase (up to 40%) in cell density, monoclonal antibody (MAb) titer, and reactor MAb productivity was observed, as compared with a conventional continuous suspension culture. The cell viability and the specific MAb productivity remained practically constant at different dialysis rates. This finding suggests that the steady state growth and death rate in continuous suspension hybridoma cultures are not direct functions of the nutrient or inhibitor concentrations.
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  • 62
    ISSN: 0006-3592
    Keywords: Fusarium graminearum ; continuous culture ; chemostat ; morphological mutants ; selection coefficient ; Ks value ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Highly branched (colonial) mutants (MC1-1-, CC1-1, and C106) of Fusarium graminearum A3/5 were each grown with the parental strain (A3/5) in continuous flow cultures at high and low dilution rates using a variety of nutrient limitations. MC1-1 replaced A3/5 in all nutrient-limited cultures tested (glucose-, Mg2+-, ammonium-, and sulphate-limited cultures), suggesting that it has a higher maximum specific growh rate than A3/5. Compared with A3/5, C106 was positively selected for in Mg2+-limited cultures and its selection coefficient was higher at low than at high dilution rates, suggesting that, compared with A3/5, it has a reduced saturation constant (Ks) for Mg2+. However, in batch culture, C106 and A3/5 had the same (15 μM) appaent Ks value for Mg2+. C106 was replaced (negative selection coefficient) by A3/5 in gluose-, ammonium-, and phsophate-limited continuous flow cultures, but was neither at an advantage nor a disadvantage (i.e., it behaved as a neutral mutation) in sulphate-limited cultures. CC1-1 replaced A3/5 when they were grown together in glucose-, maltose-, or ribose-limited continuous flow cultures, but not in fructose-, xylose-, ammonium-, or phsophate-limited cultures. Because A3/5 and CC1-1 had similar Km values (30 μM) for glucose, and because the selective advantage of CC1-1 was maintained in maltose-limited cultures (maltose was not hydrolyzed extracellularly), it was concluded that the selective advantage of CC1-1 did not result from it having a lower Ks for glucose than the parental strain. Rather, the data suggested that the activity of phosphoketopentoepimerase may be altered by the CC1-1 mutation. © 1992 John Wiley & Sons, Inc.
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  • 63
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    Biotechnology and Bioengineering 41 (1993), S. 937-946 
    ISSN: 0006-3592
    Keywords: protein excretion ; continuous culture ; Escherichia coli ; β-lactamase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The stable continuous overproduction of a plasmidencoded protein, β-lactamase, for at least 50 days by Escherichia coli K-12, RB791(pKN), with release into the culture medium has been demonstrated in two-stage chemostats. The second-stage culture was continuously induced with 0.1 mM IPTG. Continuous expression of β-lactamase could not be sustained with this strain in a single-stage chemostat because of cell death and selection for lac-1 cells. β-Lactamase production in the second stage was sensitive to the second-stage dilution rate and the distribution of the limiting substrate (i.e., glucose) between the first and second stages. The fraction of viable, excreting cells and the average copy number in the induced culture was measurably higher under those conditions of dilution rate and substrate distribution which yielded high β-lactamase levels. The best operating conditions found at 20°C were a first-stage dilution rate of 0.12 h-1, a second-stage dilution rate of 0.03 h-1, and equal glucose feed supplied to each stage. Enzymatically active β-lactamase was produced at a level of 25% of total cellular protein with 90% excretion yielding 300 mg β-lactamase/L that was 50% pure at an OD600 〈 6. © 1993 Wiley & Sons, Inc.
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  • 64
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    Biotechnology and Bioengineering 42 (1993), S. 1046-1052 
    ISSN: 0006-3592
    Keywords: fungus ; rheology ; morphology ; continuous culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Aspergillus niger was grown in a 7-L chemostat at biomass levels of 7 to 9 gL-1; dilution rates of 0.03, 0.05, 0.075, and 0.009 h-1; and dissolved oxygen tensions of 7%, 12%, and 40% of air saturation. Broth rheological measurements were made on-line, while off-line image analysis was used to measure mycelial morphology, including characterization of mycelial aggregates (clumps). Under all conditions, more than 87% of the hyphase were in clumps, the shape of which determined the rheological characteristics of the broth. In particular, the power law consistency index could be correlated with the biomass concentration and the roughness factor of the clumps, which describes their hairiness. A decrease in specific growth rate decreased roughness, possibly due to changes in the amount of clump breakup. However, decreases of roughness with increasing dissolved oxygen tension might rather imply some effect on hyphal-hyphal interactions within the clumps. © 1993 John Wiley & Sons, Inc.
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  • 65
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    Biotechnology and Bioengineering 40 (1992), S. 1218-1232 
    ISSN: 0006-3592
    Keywords: methanotroph ; biofilm ; fluidized-bed ; attached-film ; film thickness ; film density ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The feasibility of using methanotrophs in an attached-film, fluidized-bed (MAFFB) reactor system has been under investigation since 1987. Mixed culture, methane-utilizing attached biofilms were developed on diatomaceous earth particles and on granular activated carbon. The required feed gases, methane and oxygen, were supplied to the attached biofilm in disolved form using separate gas-liquid aeration columns. Biofilm growth was steady despite low influent dissolved methane concentrations (1 to 3 mg/L). A breeder MAFFB operated consistently for 4.1 years with attached biofilm concentrations as high as 51.7 g VS/L static-bed with minimal biomass wasting and with minimal buffer and nutrient inputs. The maximum biomass concentration observed was 75.6 g VS/L static-bed in a MAFFB reactor treating trichloroethene. Biofilm thickness reached 160 μm with typical values of 70 μm under methane and oxygen growht-rate-limited conditions. Biofilm densities of 120 to 190 g VS/L film were observed. Growth rates varied from 〈0.01/d to 0.17/d. Greater than 90% of the biomass concentration in the bed was attached, and effluent total suspended solids ranged from 5 to 74 mg/L, with an average of 24 mg/L over 27 runs in four MAFFB systems at upflow velocities of 11.4 to 25 m/h. Heterotrophic attached-film methanotrophs appear to be stable and useful for applications in toxics treatment, and other product manipulations. © 1992 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 479-488 
    ISSN: 0006-3592
    Keywords: acidogenesis kinetics ; lactose ; lactose ; biofilm ; mass transfer resistance ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The intrinsic fermentation kinetics of lactose in acidogenic biofilms were investigated in situ in a continuous flow fermentor at 35°C and pH 4.6. The external and internal mass transfer resistances to lactose molecules from bulk solution to inside the biofilms were experimentally minimized or eliminated in a thin biofilm and recycled medium. In a chemically defined culture medium, the immobilized acidogens converted lactose mainly to acetate and butyrate; the minor products included ethanol. propionate, lactate, and hydrogen. The utilization rate of lactose, as a function of lactose concentration in the fermentor, can be described by a Michaelis-Menten equation, as can the formation rates of acetate, butyrate, and ethanol. The production rates of propionate and lactate had a liner relationship with lactose concentration under the experimental conditions. The low pH (4.6) of culture medium could depress the formation of propionate, and intermediate which is most difficulty digested by acetogenic bacteria located in the second fermentor in a two-phase process. Production rate of acetate quickly reached a constant, and additional utilization of lactose produced more butyrate and other minor products. © 1993 John Wiley & Sons, Inc.
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  • 67
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    Biotechnology and Bioengineering 44 (1994), S. 586-594 
    ISSN: 0006-3592
    Keywords: sulphate-reducing bacteria ; biofilm ; granulation ; gas-lift reactor ; hydrogen sulphide toxicity ; mass transfer ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Feasibility and engineering aspects of biological sulphate reduction in gas-lift reactors were studied. Hydrogen and carbon dioxide were used as energy and carbon source. Attention was paid to biofilm formation, sulphide toxicity, sulphate conversion rate optimization, and gasliquid mass transfer limitations. Sulphate-reducing bacteria formed stable biofilms on pumice particles. Biofilm formation was not observed when basalt particles were used. However, use of basalt particles led to the formation of granules of sulphate-reducing biomass. The sulphate-reducing bacteria, grown on pumice, easily adapted to free H2S concentrations up to 450 mg/L. Biofilm growth rate then equilibrated biomass loss rate. These high free H2S concentrations caused reversible inhibition rather than acute toxicity. When free H2S concentrations were kept below 450 mg/L, a maximum sulphate conversion rate of 30 g SO42-/L · d could be achieved after only 10 days of operation. Gas-to-liquid hydrogen mass transfer capacity of the reactor determined the maximum sulphate conversion rate. © 1994 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 44 (1994), S. 595-608 
    ISSN: 0006-3592
    Keywords: biofilm ; aerobic waste water treatment ; airlift reactor ; waste water ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this article, the conditions for aerobic biofilm formation on suspended particles, the dynamics of biofilm formation, and the biomass production during the start-up of a Biofilm Airlift Suspension reactor (BAS reactor) have been studied. The dynamics of biofilm formation during start up in the biofilm airlift suspension reactor follows three consecutive stages: bare carrier, microcolonies or patchy biofilms on the carrier, and biofilms completely covering the carrier. The effect of hydraulic retention time and of substrate loading rate on the formation of biofilms were investigated. To obtain in a BAS reactor a high biomass concentration and predominantly continuous biofilms, which completely surround the carrier, the hydraulic retention time must be shorter than the inverse of the maximum growth rate of the suspended bacteria. At longer hydraulic retention times, a low amount of attached biomass can be present on the carrier material as patchy biofilms. During the start-up at short hydraulic retention times the bare carrier concentration decreases, the amount of biomass per biofilm particle remains constant, and biomass increase in the reactor is due to increasing numbers of biofilm particles. The substrate surface loading rate has effect only on the amount of biomass on the biofilm particle. A higher surface load leads to a thicker biofilm.A strong nonlinear increase of the concentration of attached biomass in time was observed. This can be explained by a decreased abrasion of the biofilm particles due to the decreasing concentration of bare carriers. The detachment rate per biofilm area during the start-up is independent of the substrate loading rate, but depends strongly upon the bare carrier concentration.The Pirt-maintenance concept is applicable to BAS reactors. Surplus biomass production is diminished at high biomass concentrations. The average maximal yield of biomass on substrate during the experiments presented in this article was 0.44 ± 0.08 C-mol/C-mol, the maintenance value 0.019 ± 0.012 C-mol/(C-mol h). The lowest actual biomass yield measured in this study was 0.15 C-mol/C-mol. © 1994 John Wiley & Sons, Inc.
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  • 69
    ISSN: 0006-3592
    Keywords: hybridoma ; subclone ; continuous culture ; batch culture ; igG-mRNA ; biosynthetic activities ; antibody production ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: During long-term continuous culture of the hybridoma cell line 11317, a better-producing subclone (I1317-SF11), giving improved productivity, has been selected. The comparison of the original cell line (I1317-DC) with this subclone revealed that although the growth patterns of both clones were similar, both in continuous and in batch cultures, considerable differences could be seen between the clones with respect to monoclonal antibody (MAB) accumulation, MAB production rate, the levels of mRNA coding for heavy and light chains of IgG, and some metabolic activities. In continuous culture as well as in batch culture, I1317-SF11 showed increased levels of mRNA coding for kappa and gamma chains compared with I1317-DC and/or a modified ratio of the mRNA species when compared to that in I1317-DC. Using pulse experiments, it could be established that the biosynthesis of both chains was augmented in I1317-SF11. Although the kappa and gamma mRNA levels were modified or inversed for I1317-SF11, the cells always synthesized more kappa than gamma chains. The overall increase in the synthetic activity of I1317-SF11 is suggested as one reason for the considerable increase of IgG productivity and product accumulation in continuous culture as well as in repeated batch cultures. Tests concerning metabolic activity revealed that I1317-SF11 had a predominantly glycolytic metabolism independent of growth requirements, whereas for I1317-DC the metabolism became increasingly glycolytic with increased growth. The antibody yield coefficient of I1317-SF11 on glutamine was significantly higher than that of I1317-DC for the continuous culture, whereas the antibody coefficients on glucose were almost similar for both clones under the different culture conditions used. Both antibody coefficients were considerablly influenced by the specific growth rate.All these facts together lead to the conclusion that subclone I1317-SF11 uses more of the energy available, or it was the energy and/or precursors available for the synthesis and production of MAB more efficiently than the thesis and production of MAB more efficiently than the original cell line. Although the levels of mRNA coding for heavy and light chains of IgG were modified, it could be confirmed that the overall regulation of MAB-synthesis and -production occurs post-translationally and that at higher growth rates, more biosynthetic activity is diverted to biomass production. © 1994 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 47 (1995), S. 520-524 
    ISSN: 0006-3592
    Keywords: bacillus subtilis ; plasmid ; continuous culture ; CAT ; recombinant cultures ; acid formation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The feasibility of continuous production of proteins in chemostat cultures of Bacillus subtilis was investigated. An expression system consisting of the bacterium B. subtilis BR151 carrying plasmid p602/19 was used. The plasmid contains the cat (chioramphenicol acetyltrans-ferase) gene downstream of a strong vegetative T5 promoter. It was found that, at a dilution rate of 0.2 h-1 production of relatively high levels of CAT protein (about 4% ofcellular protein) can be sustained. But, experiments at a higher dilution rate of 0.4 h-1 were unproductive because of high acidformation and washout. Combination of low cell yield, which results from excessive acid formation, and low dilution rate led to a low volumetric CAT productivity. Our recent work with the nonrecombinant cells, has demonstrated that uptake of small amounts of citrate significantly reduces or entirelyeliminates the acid formation. This superior performance in the presence ofcitrate was hypothesized, based on strong experimental evidence, to be the result of a reduction in glycolysis flux through a sequence of events leading to a reduction in pyruvate kinase and phosphof- ructokinase activities, the regulatory enzymes of glycol-ysis. In this study, it is demonstrated that cofeeding of glucose and citrate substantially reduces theorganic acid formation and significantly increases the recombinant culture productivity. The combination of high specific CAT activity and cell density resulted in a total of six- to tenfold higher culture productivitywhen citrate and glucose were cometabolized than when glucose was the only carbon source. © 1995 John Wiley & Sons Inc.
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  • 71
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    Biotechnology and Bioengineering 40 (1992), S. 413-426 
    ISSN: 0006-3592
    Keywords: Bacillus polymyxa ; product profiles ; continuous culture ; bioenergetic model ; oxygen availability ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Bacillus polymyxa ferments glucose to 1-2,3 butanediol, acetoin, ethanol, acetic acid, lactic acid, and formic acid. This research investigates product formation as a function of oxygen availability. A predictive model that simulates product distribution at known oxygen transfer rates is developed on the hypothesis that, in an energy-limited environment, B. polymyxa utilizes glucose and oxygen in the most efficient manner. The efficiency of utilization of glucose and oxygen is measured in terms of the ATP yields of each oxidative pathway. The identity of the products constituting the profile at the given oxygen transfer rate is determined by comparing the ATP production and consumption rates. While the ATP generated is calculated from a knowledge of the oxygen transfer rate and ATP yields of the oxidative pathways, the ATP consumption is estimated by the Pirt expression in terms of growth- and nongrowth-associated components. The product formation rates are obtained by solving ATP and NAD balance equations. They equate the production and consumption rates of these intermediates and are derived from the pseudo-steady-state hypothesis. The model is applied to continuous culture systems that are both open and closed with respect to biomass. At a given oxygen transfer rate, dilution rate, and inlet glucose concentration, the model predicts steady-state concentrations of two dominant fermentation endproducts with the help of four parameters that can be determined from independent experiments. In contrast with earlier approaches, the experimental studies are carried out in continuous culture. Product profiles are obtained at various oxygen transfer rates, fer rates, inlet glucose concentrations, and dilution rates. The effect of pH on the relative distribution of products is also demonstrated. Results indicate that the model is fairly successful in predicting product profiles as a function of oxygen availability. © 1992 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 39 (1992), S. 865-869 
    ISSN: 0006-3592
    Keywords: cellulase ; xylose ; Trichoderma reesei ; continuous culture ; growth modeling ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Trichoderma reesei (QM 9414) produced cellulase in continuous culture, on media containing xylose (1%) supplemented with sorbose (0.3%) to induce cellulase production. Maximum cell mass of 4.54 kg/m3 occurred at pH 4.0 and a dilution rate of 0.0391 h-1 where residual substrate was 0.43 kg/m3, but no cellulase was produced. Maximum cellulase production of 0.69 FPU occurred at pH 3.5 and a dilution rate of 0.0110 h-1, where cell mass production was 2.56 kg/m3 and residual substrate was 0.15 kg/m3. Monod kinetic constants, corrected for endogenous metabolism, were 0.091 h-1, 0.469 kg/m3, 0.00923 h-1, and 0.470 kg cells/kg xylose at pH 3.5, for the maximum specific growth rate, Michaelis-Menten coefficient, endogenous metabolism coefficient, and yield coefficient, respectively. Specific growth rate fitted a maturation time model, which predicted decreasing maturation time with increasing pH.
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  • 73
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    Biotechnology and Bioengineering 41 (1993), S. 111-117 
    ISSN: 0006-3592
    Keywords: biofilm ; detachment ; model ; physiology ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A general mathematical framework for modeling biofilm detachment is presented. The approach is founded on a material balance on biomass that equates the detachment rate to the product of a detachment frequency and a detaching particle mass. The model provides a theoretical basis for deriving many of the empirical detachment rate expressions in common use and can thus lend some insight into their physical and biological significance. By allowing for variation in the detachment frequency with depth in the biofilm, the model permits derivation of detachment expressions that reflect a dependence on chemical or physiological gradients in the biofilm. Analysis of literature data sets from two different biofilm systems suggests, in both cases, that detachment is a growth-associated phenomenon. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 42 (1993), S. 111-117 
    ISSN: 0006-3592
    Keywords: biofilm ; particle ; Pseudomonas aeruginosa ; transport ; roughness ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fluorescent latex microbeads added to a Pseudomonas aeruginosa biofilm as tracers of particle movement penetrated the biofilm and remained in it much longer than predicted by a model of advective displacement due to cell growth. Beads with a nominal diameter of 1 μm that were added in the bulk fluid became distributed throughout the biofilm depth. Some microbeads penetrated to the substratum within the 24-h bead addition period. The biofilms had a mean thickness of approximately 34 μm but have been previously shown to be quite rough. Measured rates of bead release from the biofilm corresponded to first order time coefficients of 0.01-0.03 h-1. These bead release rates were approximately an order of magnitude less than the predicted time scale of advective transport, which is just the experimentally measured specific cellular growth rate of 0.15 h-1. Computer simulations of bead transport using the biofilm model BIOSIM were compared with bead release rate data and with bead position distributions within the biofilm as determined by microscopic examination of thin cross sections of embedded biofilm. The model predicted much faster release of beads from the biofilm than actually occurred. It is hypothesized that both the ability of beads to penetrate the biofilm and the unexpectedly low advective displacement velocity of particles in the biofilm were due to the rough nature of the biofilm. © 1993 John Wiley & Sons, Inc.
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  • 75
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    Biotechnology and Bioengineering 42 (1993), S. 557-570 
    ISSN: 0006-3592
    Keywords: mathematical model of cell growth ; continuous culture ; protein excretion ; β-lactamase ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simple mathematical model is developed to help explain the complex population dynamics of an Escherichia coli host-plasmid expression/excretion system for β-lactamase within single- and two-stage reactors. The model successfully integrates the individual regulatory (tac promoter induction), genetic (runaway plasmid replication), and population dynamics (culture instability) aspects of the system. The model predicts, and experiment confirms, that high-level β-lactamase production and excretion cannot be easily maintained in single-stage reactors using the current plasmid construction. Stable target protein production and excretion is mathematically predicted, and experimentally confirmed, within two-stage reactors. The model is used to provide insight into engineering a more stable host-vector expression/excretion system for use in single-stage reactors. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 42 (1993), S. 974-986 
    ISSN: 0006-3592
    Keywords: hybridoma cell culture ; batch culture ; continuous culture ; integrated product recovery ; monoclonal antibodies ; proteases ; glycosylation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The molecular integrity of monoclonal antibodies (MCAB) produced by murine hybridoma cell line TB/C3 was studied in batch and continuous-flow cultures. In batch culture, one band of MCAB was detected initially by Western blotting of sodium dodecyl sulfate (SDS)-polyacrylamide gels run under unreduced conditions, but heterogenous MCAB bands appeared as the culture aged. The latter were due to the degradation of MCAB by proteases active at the neutral pH of the culture. The deleterious effect of proteases was minimized in the continuous-flow cultures which were integrated for product recovery. The MCAB of high quality was purified over 26 days from a culture grown at a dilution rate of 0.025 h-1 (experiment 1). However, at a lower dilution rate of 0.015 h-1 (experiment 2), the integrity of MCAB was compromised after the initial 13 days of culture. This was shown to be due to the variation in the carbohydrate content of MCAB produced, as judged by the increased sialylation of heavy chains and the varied reactivity of MCAB with lectins (Maackia amurensis agglutinin, Galanthus nivalis agglutinin, and Datura stramonium agglutinin) as the age of the culture increased. The concentration of the purified MCAB samples by enzyme-linked immunosorbent assay (ELISA) (used normally) was usually higher than that estimated by absorbance at 280 nm. Best correlation between the two methods (ELISA-280 nm ratio of 1.02-1.25) was obtained with experiment 1 samples. This ratio increased in experiment 2 and batch culture samples as the heterogeneity of MCAB produced increased, being 1.03-2.94 and 2.53-4.62, respectively. Therefore, ELISA overestimated MCAB concentration when the molecular integrity of the latter was compromised. The ELISA-A280 nm ratio might hence provide a useful indicator for assessing the quality of MCAB produced. Comparison of SDS-polyacrylamide gels stained with Coomassie Brilliant Blue R and silver showed that the former correlated better with the MCAB activity stain, whereas the silver stained both the protein- and carbohydrate-rich components. Comparison of the patterns produced with these two stains might therefore offer another parameter to monitor the overall integrity of MCAB produced. Finally, the data presented have important implications on the validity of using long-term and intensive cultures for generating MCAB because such cultures would be subjected to the additive effects reported for batch and continuous modes of growth. © 1993 John Wiley & Sons, Inc.
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  • 77
    ISSN: 0006-3592
    Keywords: biofilm ; extracellular biopolymer ; lead microbe interaction ; metal toxicity ; structured models ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The interfacial interactions of a toxic trace metal, Pb, with a surface modified by a marine film-forming bacterium, Psedomonas atlantica, were predicted by a structured biofilm model used in conjunction with a chemical speciation model. The validity of the integrated model was tested for batch and continuous operations. Dynamic responses of the biophase due to transient lead concentration increases were also stimulated. The reasonable pre dictions achieved by the model demonstrate its utility in describing trace metal distributions in complex systems where the adsorption properties of inorganic surfaces are modified by adherent bacteria production of extracellular polymers. © 1994 John Wiley & Sons, Inc.
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  • 78
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    Keywords: biofilm ; structured models ; extracellular biopolymer ; microbial attachment/detachment ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Adsorbent surfaces in natural and engineered systems are frequently modifies by bacterial attachment, growth of a biofilm, and bacterial production of extracellular polymer. Attached cells or sorbed polymers may alter the metal-binding characteristics of the supporting substratum and influence metal partitioning. The interdependent behavior of toxic trace metal partitioning and biofilm development requires description of the interaction between cell growth with its accompanying polymer production and metal speciation. In this article, the first of a two part series, a mechanistic model is developed to describe the growth of a film-forming bacterium which adheres to a substratum through the production of extracellular biopolymers. Each bacterial cell was modeled as a two-component structure consisting of active cell mass and biopolymer. The biopolymer component was further divided into cell-associated and dissolved categories to distinguish which remained naturally bound to cell surfaces from that which did not. Use of this structured model permitted independent description of the dynamics of cell growth, and polymer production, both of which may influence trace metal behavior. Employing parameters obtained from independent experiments as well as published values, the model satisfactorily predicts experimental observations of bacterial growth, attachment and detachment, biopolymer production, and adsorption of polymer onto solid (glass) surfaces. The model stimulated transient and steady-state biofilm systems equally well. In the second article in this series, we describe how this model may be extended and utilized to make predictions of the behavior of transient and steady-state biofilm systems in the presence of a toxic transition metal(Pb). © 1994 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 43 (1994), S. 471-476 
    ISSN: 0006-3592
    Keywords: hexavalent chromium ; microbial reduction ; Pseudomonas sp. ; continuous culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Reduction of hexavalent chromium was studied in three bench-scale continuous stirred tank reactors. The inoculum was a culture of Pseudomonas sp., capable of giving 83% to 87% chromate reduction in 72-h batch assays with 60 mg Cr(VI) L-1 in synthetic medium. The continuous culture studies were conducted for about 100 days using synthetic feed containing different levels of chromate (5 to 124 mg L-1) at 28° to 30°C and pH 6.8. The feed rate was varied over the range 0.5 to 1 L d-1 to obtain hydraulic retention time of 36 to 72 h. Chromate reduction efficiency was 81% to 91% and 100% for influent Cr(VI) concentrations of 15 to 124 and 5 mg L-1, respectively, with a hydraulic retention time of 72 h. © 1994 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 44 (1994), S. 1279-1287 
    ISSN: 0006-3592
    Keywords: waste gas ; trickling filter ; biofilm ; dichlo-romethane ; biofiltration ; air pollution ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Clogging is well-known phenomenon in the application of a biological tricking filter for both waste gas and wastewater treatment. Nevertheless, no such observations or even significant changes in pressure drop have ever been recorded during the long-term processing of a waste gas containing dichloromethane (DCM) as a sole carbon source. To obtain more information about this phenomenon, a detailed investigation into the carbon balance of this system has been performed. During a period of operation of about 200 days the rate of DCM elimination and the overall rate of CO2 production in a continuously operating filter were therefore recorded daily, thus allowing an evaluation of the overall conversion process. Furthermore pseudo-steady-state measurements were carried out on a regular basis. These experiments reveal more detailed information on the actual DCM conversion by Hyphomicrobium GJ21 within the biofilm. The combined results of the experiments described in this article show that on an overall basis a so-called biological equilibrium, i.e., a situation of no net biomass accumulation, is obtained in the course of time. It appeared that the overall rate of CO2 production slowly increased until, after some 200 days, it finally counter-balanced the conversion rate of DCM on a molar-basis. As opposed to this result, all pseudo-steady-state experiments indicated that about 60% of the eliminated primary carbon source is converted into biomass. This is in good agreements with results from microkinetic experiments. Based on these results and evaluation of the experimental data, it is concluded that interactions between several microbial populations are involved in this biological equilibrium. These interactions include both biomass growth and biomass degradation. © 1994 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 47 (1995), S. 585-595 
    ISSN: 0006-3592
    Keywords: biofilm ; wastewater treatment ; airlift reactor ; nitrification ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: For a stable and reliable operation of a BAS-reactor a high, active biomass concentration is required with mainly biofilm-covered carriers. The effect of reactor conditions on the formation of nitrifying biofilms in BAS-reactors was investigated in this article. A start-up strategy to obtain predominantly biofilm-covered carriers, based on the balancing of detachment and a biomass production per carrier surface area, proved tp be very successful. The amount of biomass and the fraction of covered carrier were high and development of nitrification activity was fast, leading to a volumetric conversion of 5 kgN · m-3 · d-1 at a hydraulic retention time of 1h. A 1-week, continuous inoculation with suspended purely nitrifying microorganisms resulted in a swift start-up compared with batch addition of a small number of biofilms with some nitrification activity. The development of nitrifying biofilms was very similar to the formation of heterotrophic biofilms. In contrast to heterotrophic bio-films, the diameter of nitrifying biofilms increased during start-up. The detachment rate from nitrifying biofilms decreased with lower concentrations of bare carrier, in a fashion comparable with heterotrophic biofilms, but the nitrifying biofilms were much more robust and resistant. Standard diffusion theory combined with reaction kinetics are capable of predicting the activity and conversion of biofilms on small suspended particles. © 1995 John Wiley & Sons Inc.
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    Biotechnology and Bioengineering 38 (1991), S. 1271-1279 
    ISSN: 0006-3592
    Keywords: PL promoter ; plasmid content ; 45S RNA ; gene expression ; continuous culture ; two-stage ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In order to investigate how plasmid content, transcription efficiency, and translation efficiency affect the productivity of a cloned gene protein, a new vector (pPLc-RP4.5) was constructed. The vector has PL promoter and lacZ as a structure gene 4.5S RNA gene between PL promoter and lacZ gene. We took advantage of the characteristic that the 4.5S RNA is accumulated inside E. coli cells and can be quantitatively measured. A two-stage continuous culture system in combination with a temperature-sensitive gene switching system was used to study the performance of the recombinant fermentation. It was found that the plasmid content as varied by the dilution rate in the production stage showed a different pattern from that in the growth stage. The result showed that promoter strength had a greater influence on the overall gene expression efficiency of a cloned gene than the plasmid content, and the overall gene expression efficiency was largely dependent upon translation efficiency when a multicopy plasmid (pBR322 derivative and rop-) and a strong promoter (PL) were used to express a heterologous protein in E. coli.
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    Biotechnology and Bioengineering 40 (1992), S. 1243-1255 
    ISSN: 0006-3592
    Keywords: calorimetry, Saccharomyces cerevisiae ; Kluyveromyces fragilis ; yeast metabolism ; modeling ; continuous culture ; respiratory capacity ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Yeasts degrade glucose through different metabolic pathways, where the choice of the pathway is dependent on the nature of the limitation in the various substrates. When oxygen is limiting in addition to glucose, yeasts often grow according to a mixture of oxidative and reductive metabolism. Oxygen may be limiting either by supply or by inherent biological restrictions such as the respiratory bottleneck in Saccharomyces cerevisiae or by both. A unified model incorporating both supply and biological limitations is proposed for the quantitative prediction of growth rates, consumption and production rates, as well as key metabolite concentrations during mixed oxidoreductive metabolism occuring as a result of such oxygen limitations. This simple unstructured model can be applied to different yeast strains while at the same time requiring a minimum number of measured parameters. “Estimators” are utilized in order to predict the presence of supply-side or biological limitations. The values of these estimators also characterize the relative importance of oxidative to total metabolism. Results from the aerobic and oxygen-limited chemostat cultures were used to corroborate the model predictions. During these experiments, the heat released by the yeast cultures was also monitored on-line. The model correctly predicted the overall stoichiometry, steady-state concentrations, and rates including heat dissipation rates measured in the various situations of oxygen limitations. Direct continuous measurements such as heat can be used in conjunction with the unified model for on-line proces control. © 1992 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 40 (1992), S. 1293-1299 
    ISSN: 0006-3592
    Keywords: on-line rheology ; continuous culture ; fungal fermentation ; dissolved oxygen tension ; specific growth rate ; biomass level ; rheology ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The influence of oxygen tension, biomass concentration, and specific growth rate on the rheological properties of an Aspergillus niger fermentation broth was investigated by growing the fungus in continuous culture. The rheological properties were measured on-line using an impeller rheometer system. The effect of the specific growth rate on broth rheology was strongly influenced by the dissolved oxygen, (DO) concentration in the broth. At DO concentrations above 10% of saturation, K increased with the dilution rate, and at DO concentrations below 10% of saturation, K decreased with increasing dilution rate. The largest influence of a change in the DO concentration on the viscosity of the broth was found at the lowest growth rates and the lowest DO concentrations. K/x, a term that gives a simple description of the structure or the morphology of the culture, was found to increase with biomass concentration. © 1992 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 211-220 
    ISSN: 0006-3592
    Keywords: plasmid retention ; gene expression ; biofilm ; β-galactosidase ; segregational instability ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Differences in plasmid retention and expression are studied in both suspended and biofilm cultures of Escherichia coli DH5α(PMJR1750). An alternative mathematical model is proposed which allows the determination of plasmid loss probability in both suspended batch and continuously fed biofilm cultures. In our experiments, the average probability of plasmid loss of E. coli DH5α(pMJR1750) is 0.0022 in batch culture in the absence of antibiotic selection pressure and inducer. Under the induction of 0.17 MM IPTG, the maximum growth rate of plasmid-bearing cells in suspended batch culture dropped from 0.45 h-1 to 0.35 h-1 and the β-galactosidase concentration reached an experimental maximum of 0.32. pg/cell 4 hours after the initiation of induction. At both 0.34 and 0.51 mM IPTG, growth rates in batch cultures decreased to 0.16 h-1, about 36% of that without IPTG, and the β-galactosidase concentration reached an experimental maximum of 0.47 pg/cell 3 hours after induction.In biofilm cultures, both plasmid-bearing and plasmid-free cells in increase with time reaching a plateau after 96 hours n the absence of both the inducer and any antibiotic selection pressure. Average probability of plasmid loss for biofilm-bound E. coli DH5β(pMJR1750) population was 0.017 without antibiotic selection. Once the inducer IPTG was added, the concentration of plasmid-bearing cells in biofilm dropped dramatically while plasmid-free cell numbers maintained unaffected. The β-galactosidase concentration reached a maximum in all biofilm experiments 24 hours after induction; they were 0.08, 0.1, and 0.12 pg/cel under 0.17, 0.34, and 0.51 mM IPTG, respectively. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 39 (1992), S. 914-922 
    ISSN: 0006-3592
    Keywords: biofilm ; competition ; modeling ; multispecies ; nitrification ; species distribution ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A steady-state model for quantifying the space competition in multispecies biofilms is developed. The model includes multiple active species, inert biomass, substrate utilization and diffusion within the biofilm, external mass transport, and detachment phenomena. It predicts the steady-state values of biofilm thickness, species distribution, and substrate fluxes. An experimental evaluation is carried out in completely mixed biofilm reactors in which slow-growing nitrifying bacteria compete with acetate-utilizing heterotrophs. The experimental results show that the model successfully describes the space competition. In particular, increasing acetate concentrations causes NH4+-N fluxes to decrease, because nitrifiers are forced deeper into the biofilm, where they experience greater mass-transport resistance.
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    Biotechnology and Bioengineering 39 (1992), S. 1069-1079 
    ISSN: 0006-3592
    Keywords: carbon dioxide ; bicarbonate ; alkalophilic cultures ; nonideal solutions ; continuous culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The chemical reactions involving carbon dioxide in mineral culture media are considered. A mathematic model is set up, based on published data, which is valid at pH values below 9, and in which the nonideality of the solution is taken into account. The crucial parameter is the constant expressing the equilibrium between carbon dioxide and bicarbonate, K1.The reactions were studied in three different aqueous solutions: water, mineral salt medium, and a suspension with nongrowing bacterial cells. For each situation, three methods were compared for the determination of the bicarbonate concentration in the solution: equilibrium state total carbon analysis, dynamic monitoring of the rate of acid or alkali addition, and dynamic measurement of the carbon dioxide gas phase mole fraction.In a batch-stirred tank reactor, the equilibrium constant K1 agreed with the published value, and the three bicarbonate analysis methods give the same results. If the nonideality is not taken into account, the result significantly differed from the published value and is likely to be incorrect.A real alkalophilic process, using Acinetobacter calcoaceticus in a continuous stirred tank reactor at steady state, also gave results that are in accord with the literature. However, the results do not allow validation of the equation expressing the nonideality.The steady state in the batch system and in continuous culture can be well described with the mathematical model. However, in the transient state there are some unexplained differences between simulation and measurement.
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  • 88
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 39 (1992), S. 1141-1146 
    ISSN: 0006-3592
    Keywords: biofilm ; diffusion ; diffusivity ; immobilized cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An experimental reactor consisting of two chambers, separated by a porous ceramic immobilization matrix, was constructed to measure the effective diffusivity of different compounds and the consumption rates of acetate in developing biofilms. In initial experiments, effective diffusivities for acetate, propionate, isopropanol, and lithium salt through the ceramic immobilization matrix in the absence of biofilm were determined to be 40% to 50% less than in water at infinite dilution. The effective diffusivity of the lithium salt was similar to that of acetate. The effective diffusivity of the lithium salt through biofilms of thickness in the range of 200 to 1200 μm was essentially constant with a value of approximately 7% of that in water at infinite dilution. Acetate consumption in the biofilm was linearly proportional to biofilm thickness up to a biofilm depth of 800 μm. Deviation from linearity appeared in biofilm thicknesses greater than 800 μm. Results of these experiments support previous reports that immobilized cell reactors have significantly higher bioconversion rates than suspended cell systems.
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  • 89
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 44 (1994), S. 636-641 
    ISSN: 0006-3592
    Keywords: biofilm ; hydrodynamics ; mass transport ; particle tracking ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Liquid flow was studied in aerobic biofilms, consisting of microbial cell clusters (discrete aggregates of densely packed cells) and interstitial voids. Fluorescein microinjection was used as a qualitative technique to determine the presence of flow in cell clusters and voids. Flow velocity profiles were determined by tracking fluorescent latex spheres using confocal microscopy. Liquid was flowing through the voids and was stagnant in the cell clusters. Consequently, in voids both diffusion and convection may contribute to mass transfer, whereas in cell clusters diffusion is the dominant factor. The flow velocity in the biofilm depended on the average flow velocity of the bulk liquid. The velocity profiles in biofilms were linear and the velocity was zero at the substratum surface. The velocity gradients within biofilms were 50% of that near walls without biofilm coverage. The influence of the biofilm roughness on the flow velocity profiles was similar to that caused by rigid roughness elements. © 1994 John Wiley & Sons, Inc.
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  • 90
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 47 (1995), S. 696-702 
    ISSN: 0006-3592
    Keywords: Bacillis subtilis ; spore mutant ; fed-batch ; continuous culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: To alleviate plasmid instability and to prolong the production phase of subtilisin, integrable plasmid and spore mutants are used. Compared with batch-type shake flask cultures, spore mutants' ability to produce subtilisin can be well pronounced in fed-batch and continuous cultures. Hence, the two culture methods make it possible to identify the peculiar characteristics of the spore mutants unobtainable in batch culture. Spore mutants can enhance subtilisin productivity and prolong subtilisin production time in fed-batch culture as well as enable us to use very low dilution rates (〈0.1 h-1) without losing productivity in continuous culture, thereby improving the conversion yield of the nitrogen source. At 0.05 h-1 the spollG mutant of Bacillus subtilis DB104 (Δnpr Δapr) (Emr) spollG (Bimr):: pMK101 (Cmr) showed a subtilisin yield about ten times higher than that from wild-type DB104 (Δnpr Δapr)::pMK101 (Cmr). © 1995 John Wiley & Sons, Inc.
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  • 91
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The variation in the behaviour of bulk solids as a result of sudden pressure changes in the surrounding gas phase is of interest in the application of lock hoppers for transfer of bulk solids into pressure vessels. By using such sluice systems, problems occur due to consolidation of the pressurized bulk solids. This paper presents the results of experimental and theoretical research, which allow to describe the influence of several parameters on pressure compensation and the degree of consolidation. With reference to the forces and mechanisms responsible for consolidation, it is possible to describe the different consolidation behaviour of incompressible (limestone) and compressible (hard coal) bulk solids.
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  • 92
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    Weinheim : Wiley-Blackwell
    Chemical Engineering & Technology - CET 14 (1991), S. 65-72 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A study was made of the flow characteristics of large oscillating drops of pure liquid-liquid systems, using a thermostatically-controlled, rising drop column, 50 mm in diameter and 1000 mm in length. Mirrors in the jacket enabled front and side views of drops to be photographed simultaneously. Single drops in the size range 5-10 mm were investigated with both mutually-saturated phases and when the solute was being transferred from the dispersed phase. The systems studied were (1) toluene and acetone (dispersed)-water (continuous), and (2) n-heptane and acetone (dispersed)-water (continuous). Acetone concentrations were varied up to 3.75 kmol/m3. The oscillations of a travelling drop were asymmetrical; therefore, the amplitude cannot be expressed accurately in terms of only two axes. The area change of the drop compared to that of a sphere of equal volume ‘ε’, was shown to represent the amplitude accurately. The periods of droplet oscillation were uniform for the mutually saturated systems of constant physical and flow properties but changed when mass transfer was taking place. The interfacial tension exerted a marked effect on the amplitude, which also depended upon the oscillation frequency. The amplitude changed with droplet size in a similar manner to the terminal velocity, i.e. it increased with increasing size until it reached a maximum, subsequently decreasing less rapidly. The drag coefficient increased with increasing rate of mass transfer from the drop. Correlation of the results and the area eccentricity ‘ε’ by dimensional analysis embracing all possible parameters and physical properties affecting drop oscillation, resulted in the correlation ε = 0.22 Sr0.42 We-0.53 M0.13 with a mean deviation of ± 14%. This will facilitate more accurate prediction of the interfacial area for mass transfer calculations, relating to equipment containing droplets in the oscillating regime.
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  • 93
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    Weinheim : Wiley-Blackwell
    Chemical Engineering & Technology - CET 14 (1991), S. 113-117 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The reduction of nitric oxide with ammonia on iron oxide catalysts has been studied in a continuous-flow recycle reactor using simulated flue gas in the temperature range from 573 to 673 K. NO and HN3 concentrations were varied between 0 and 1000 vpm, O2 and H2O concentrations between 0 and 9 vol.-%, the remainder being nitrogen. In the presence of oxygen, the formulated reaction rate equation describes the measured rates of the main reaction NO + 2/3 NH3 ⇄ 5/6 N2 + H2O. Its form corresponds to the Langmuir-Hinshelwood type. The rate equation well fits the data, which cover the whole industrial temperature and concentration range. In the absence of oxygen, the measured reaction rates can be best described by a power law.
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  • 94
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new transport system has been developed which combines vibration of the conveying pipe with amplitudes of the order of fractions of a millimetre with rather low superficial velocities of the carrier gas. This new system was specially designed for gentle transport of fragile particulate materials.
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  • 95
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    Weinheim : Wiley-Blackwell
    Chemical Engineering & Technology - CET 14 (1991), S. 147-161 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this study, we investigated the limits of safe operation for a cooled reactor, operated batchwise. As an example of a single-phase reaction, we studied the decomposition of t-butyl peroxypivalate, a well-known organic peroxide, undergoing self-heating at relatively low temperatures. If sufficiently diluted, it can be supplied to a polymerization process from large, cooled but unstirred vessels. We present a number of extensions to the existing homogeneous explosion theory, namely a practical definition of the critical condition, its calculation, and expressions for the available time before runaway in the case of a supercritical condition, taking into account the effects of natural convection inside the vessel and the reactant conversion. The extensions of the theory were confirmed by adiabatic and non-adiabatic runaway experiments on bench scale, and natural convection cooling experiments with liquids in various packages.
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  • 96
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This contribution deals with the consequences of the decrease in diffusion coefficients at the boundaries of miscibility in fluid systems and its effect on the mechanisms of mass transfer. The liquid-liquid (1-1) extraction involving several components is taken as an example for the discussion of this topic. A short review of Hampe's theory on transport mechanisms in the transition regions of interfaces [1] explains the intrinsic interdependence between mass and momentum transfer. Finally, a discussion of the peculiarities of resistances to mass transfer in boundary layers, based on the changes in molecular transport properties, is presented.
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  • 97
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    Weinheim : Wiley-Blackwell
    Chemical Engineering & Technology - CET 14 (1991), S. 200-208 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The catalytic properties of wide-pore zeolites, namely Ni- and Cu-doped faujastie H-Y and H-mordenite (H-M) are compared on the example of conversion of n-hexane. After thermal pretreatment of ion-exchanged zeolites, reduction with hydrogen (PH2 ≤ 40 bar) leads to a large number of metallic clusters and crystallites as well as ionic species in the zeolites. Notwithstanding this variety, the different types of catalysts can be divided into three classes: classical bifunctional catalysts (for example NiHM, NiHY) with ion exchange degrees 〉 30%, monofunctional catalysts, such as CuHM and CuHY with high copper contents (ion exchange degree 〉 40%) and metal-doped (Ni and Cu) zeolites which, depending on the conditions of pretreatment, act as bifunctional and/or monofunctional compounds and, therefore, show very unstable reaction behaviour. In addition to the extent of reduction, the cation distribution affects the activity and deactivation behaviour and, to a lesser extent, the product selectivity. The reaction mechanism of bifunctional catalysis is similar to that observed in platinum-doped catalysts, while the mechanism of monofunctional catalysis resembles the process on undoped zeolites.
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  • 98
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This work investigates the thermodynamically based assumption that diffusion at binodal states of unstable fluid systems vanishes. It is shown experimentally that a scaling law, which describes the fall in diffusion near critical points, may also be applied in an extended form to its abatement at the limits of solubility. These results may profoundly affect the modelling of mass transfer between phases whose bulks exhibit states of saturation.
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  • 99
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    Weinheim : Wiley-Blackwell
    Chemical Engineering & Technology - CET 14 (1991), S. 240-247 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A systematic method of controller design is introduced to determine the overall charcteristic behaviour developed on process pole removal. The time delay compensation is automatically incorporated in the proposed control law. The implemented tuning parameters in the law are confined to the range between 0 and 1 to guarantee the stability of the overall control system. Subsequently, the fixed and adaptive control strategies are implemented to simulate a batch PVC reaction system. The adaptive control scheme provides good, roubust control of this simulated reactor notwithstanding the wide range of operating conditions and non-linear dynamics of the system. However, the fixed control scheme performs well only for a noise-free system. In addition, two limiting control laws, derived from the proposed method, are also used to simulate the reactor. The results indicate that these laws are not suitable for this non-linear reaction system.
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  • 100
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article describes the effects which occur during three-component mass transfer in liquid-liquid extraction between saturated and undersaturated phases of the system glycerol-acetone-water. The description is based on experiments in a countercurrent spray column. The physical modelling starts with the balance equations of phase flows, taking into account the variation of concentrations and mass flow rates along the column axis as well as backmixing effects. In addition, recent findings on the mechanisms of mass transfer in saturated and undersaturated phasses, deduced from theoretical and experimental results, were used to formulate mass transfer coefficients. These are valid for the region near the interface of drops in a fluid and are inserted into the balance equations. A comparison between experiments and calculations of the observed phenomena is presented.
    Additional Material: 15 Ill.
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