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  • Biochemistry and Biotechnology  (3,182)
  • Wiley-Blackwell  (3,182)
  • 1990-1994  (3,182)
  • 1
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 35 (1990), S. 8-14 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A fast inferential, multivariable adaptive optimization algorithm based on a fast responding off-gas data, the carbon dioxide evolution rate (CER), has been developed and applied to a continuous baker's yeast culture to maximize the cellular productivity in simulation and experimental studies. In the simulation study the process was optimized based on CER measurements using readily available steady-state data on the ratio between the cellular productivity and the CER. It was shown that the algorithm is two to three times faster than the algorithm based on cell mass concentration measurements. In the experimental study the CER was maximized without any information on the relationship between the cellular productivity and the CER. It took about 40 h for the process to converge, while about 80 h was required when the optimization was based on cell mass measurements. The attained steady state was found to be different but fairly close to that obtained with cell measurements. Briefly discussed is a switching to the cell-mass-based algorithm at the final stage of the optimization to overcome a potential inaccuracy.
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  • 2
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    Biotechnology and Bioengineering 35 (1990), S. 43-49 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Measurements of kLa were carried out in 1. 5- and 5-L New Brunswick Scientific CelliGen® bioreactors. The measured kLa in water were identical for both vessel sizes operated in similar condition. The mass transfer rate increased with temperature, mixing speed, and aeration rate, with this last parameter being the most significant. Surface aeration alone gave kLa values of 0. 4 to 1. 6 h-1. A 25% decrease in kLa was observed above an aeration rate of 1. 6 vvm. This was caused by the particular foam breaker of the CelliGen bioreactor. Measurements of kLa using a mammalian cell culture medium supplemented with 5% fetal calf serum (FCS) have confirmed the negative effect of the foam breaker on kLa The measured value in this medium was 1. 2 h-1 for all aeration rates, more than 60% of which was attributed to surface aeration.
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  • 3
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    Biotechnology and Bioengineering 35 (1990), S. 94-98 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 4
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    Biotechnology and Bioengineering 35 (1990), S. 103-107 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 5
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 35 (1990), S. 73-86 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A major cost consideration in the use of anaerobic digestion to convert biomass and waste to utility-grade gas is the expense of separating CO2 from the product gas. Anaerobic digestion has a number of inherent properties that can be exploited to increase the methane content of the gas directly produced by the digester, the most important of which is the high solubility of CO2(40-60 times that of methane) in water under digestion conditions. The methane enrichment concept examined in this study involved the recirculation of a liquid stream from the digester through a CO2 desorption process and the return of the liquid stream back to the digester for absorption of additional CO2 produced by the conversion of organic materials. A steady-state equilibrium model predicted that a digester gas methane content exceeding 94% could be achieved with this scheme using modest recirculation rates provided a desorption process could be designed to achieve a 60+% CO2 removal efficiency in the degassing of the liquid recycle stream. Using fixed-film laboratory digesters operated on synthetic feedstocks, the technique of methane enrichment was tested under pressurized and unpressurized conditions. A 93 + 2% methane gas stream was produced from a volatile-acid-fed bench-scale digester simulating the methanogenic stage of two-phase digestion under conditions of (1) a pH swing achieved without caustic addition that allowed digestion at pH 7. 5 and air stripping at pH 6. 5-7. 0, (2) digester pressurization to 30 psig, and (3) a recycle rate of 0. 33 L/L reactor/day. Significant but lower levels of methane enrichment were achieved with the single-stage digester at the low experimental recycle rate. However, the narrow range among all experiments of CO2 desorption efficiencies achieved in air stripping the recycle stream (35-60% CO2 removal) suggests that comparable methane enrichment-may be achieved with unpressurized single-stage digestion using greater recycle rates. A materials balance analysis of data from an unpressurized, single-stage digester employing no chemical addition and using laboratory degassing efficiencies indicated that 94% methane could be produced at recycle rates of less than 1. 4 L/L reactor/day with a methane loss of less than 2%.
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  • 6
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    Biotechnology and Bioengineering 35 (1990), S. 87-93 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A water-soluble, ligand-bound polymer has been synthesized for the purpose of isolation of urokinase, an important plasminogen activator. The affinity polymer was formed by copolymerizing N-acryloyl-m-aminobenza-midine and acrylamide in the absence of oxygen. An affinity ultrafiltration process was then developed for isolating urokinase from an artificial solution containing peroxidase and urokinase and from a crude urine source. The process yields were determined to be 86% and 49%, respectively. The recovered urokinase exhibited a specfic activity close to that of the highest commercial grade. This article also presents a new technique for assaying urokinase by coupling plasminogen with L-benzoyl arginine-p-nitroanilide (L-BAPNA), an inexpensive chromogenic substrate.
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  • 7
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: By feeding ethanol at various high rates to low cell density cultures of Saccharomyces cerevisiae it was shown that the sharp fall in viability when ethanol is produced during rapid fermentations is in part a direct consequence of the high rate of change of extracellular ethanol concentration. Nevertheless, the fall in viability in high cell density rapid fermentations which produced 98 g L-1 ethanol in 3 h considerably exceeded that of control low cell density cultures to which ethanol was added at the same rate. This difference was shown to be not due to intracellular ethanol accumulation or to differences in glucose concentration between the cultures. The concentrations of a range of potentially toxic fatty acids, higher alcohols, and esters were measured during rapid fermentations, but when added at these concentrations to control cultures in the presence of ethanol they had no significant toxic effect. However, when rapid fermentations were conducted in rich medium containing 80 g L-1 yeast extract, the apparent difference in toxicity of produced and added ethanol virtually disappeared. Magnesium was shown to be the component of yeast extract primarily responsible for this effect. The high rate of fall of viability when ethanol is rapidly produced is suggested to be partly due to the inability of the cells to adapt quickly enough to the rising ethanol concentration and partly to an increased demand for magnesium at higher ethanol concentrations which cannot be met in Mg-unsupplemented high cell density fermentations.
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  • 8
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    Biotechnology and Bioengineering 35 (1990), S. 559-564 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The method of resting cells has been of interest in the development of biocatalysts applied to organic reactions.This article deals with the use of resting cells of a thermophilic archaebacterium Sulfolobus solfataricus, in the asymmetric reduction of acyclic, cyclic, and aromatic ketones. The system allows the continuous regeneration of endogenous coenzyme with the coupled substrate approach. The results indicate that the direction of hydride attack was equatorial on the re face of the carbonyl group of substrates producing (S)-alcohols with a good optical yield. A convenient system for the reuse of resting cells has been set out to synthesize (S)-alcohols on a preparative scale.
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  • 9
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    Biotechnology and Bioengineering 35 (1990), S. 578-585 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new fiber-optic dissolved oxygen sensing technique was applied to the study of two-phase aqueous/perfluorocarbon (pfc) dispersions. These dispersions were examined for their oxygen transfer enhancement capability in the absence and presence of an oxygen-consuming reaction. For the pfc-in-water dispersions, oxygen uptake rate (OUR) enhancements were equal both with and without oxygen-consuming cells present in the aqueous phase. In contrast, for water-in-pfc dispersions, OUR enhancements inthe presence of reaction were limited by oxygen diffusion across the aqueous phase droplets. Nevertheless, enhancement factors of 5-10 on an aqueous phase volume basis were obtained in a 75% pfc dispersion.These oxygen transfer enhancements were directly translatable into enhancements in overall fermenter productivity for actual microbial cultivation systems.
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  • 10
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    Biotechnology and Bioengineering 35 (1990), S. 650-652 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 11
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 35 (1990) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 12
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    Biotechnology and Bioengineering 35 (1990), S. 99-102 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 13
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    Biotechnology and Bioengineering 35 (1990) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 14
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    Biotechnology and Bioengineering 35 (1990), S. 138-145 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pirt's maintenance model has been widely accepted for the effects of growth rate and maintenance on growth yield. However, the interpretation of parameters in Pirt's model as biological constants is difficult for energy-sufficient culture growth. In this study, a mechanistic model for the growth energetics of energy-sufficient chemostat cultures is proposed and verified with literature data. In the model, the overutilization of the energy substrate in energy-sufficient culture growth is attributed to the defective regulation of the energy substrate metabolism and energy uncoupling. The model also uses an “energy surplus” concept to collectively represent the effects of energy excessiveness. The proposed model provides a better quantitative understanding of the maximum growth yield and maintenance of energy-sufficient cultures. It also explains the glucose concentration effect reported in the literature.
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  • 15
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    Biotechnology and Bioengineering 35 (1990), S. 146-151 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of charged residues in peptide antigens on the binding characteristics of polyclonal antipeptide antibodies were studied using immunoadsorbents prepared by coupling the antibodies to CNBr-activated Sepharose 4B. Among the antipeptide antibodies, an antibody to the peptide without charged residues showed the most stable interaction with the peptide to the changes in pH. Conversely, the binding affinity of antibodies to the pep-tides with histidine residues having a unique pKa value of 6.0 decreased steeply with pH at around 6.0. The binding affinity of an antibody to the peptide with many charged residues decreased steeply with an increase in the ionic strength (adjusted by NaCl). Since circular dichroism (CD) spectrum measurements indicate that these peptides show disordered structures in the pH range of adsorption measurement, the dependence of peptide-antibody interaction on environmental conditions is attributed to the characteristics of side chains of the peptides. These results indicate that the dependence of the binding affinity of antipeptide antibodies on pH and the ionic strength is dominantly affected by the number and the pKa values of charged residues in the peptides.
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  • 16
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    Biotechnology and Bioengineering 35 (1990), S. 207-210 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 17
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    Biotechnology and Bioengineering 36 (1990), S. 28-38 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Sustained oscillations of biomass, ethanol, and ammonium concentrations, specific growth rate, and specific uptake rates of ethanol, ammonium, and oxygen were found in continuous cultures of Saccharomyces cerevisiae under controlled dissolved oxygen (DO), pH, and temperature conditions. The period of oscillations was approximately 2.5-3 h at a pH of 5.5 and 2-2.5 h at a pH of 6.5. Oscillations were observed only under conditions of low carbon (glucose below the minimum detectable level), nitrogen nutrient (ammonium concentration varied between 0.00001 and 0.0015M), and ethanol concentration (0.002-0.085 g/L) in the bioreactor.The oscillatory behavior at pH 5.5 was also characterized by partially synchronized cell growth and reproduction. Not only did the total percentage of budding cells oscillate with the same period as observed for the global biomass and nutrient concentrations, but the peaks in the individual subpopulations of initial budding, middle budding, and late budding cells appeared sequentially during the oscillation period. This provides strong evidence of the hypothesis that variations in metabolism during different periods in the cell cycle of a partially synchronized cell population are responsible for the observed oscillatory bioreactor behavior.The specific nutrient uptake rates for ammonium and oxygen as well as the net specific ethanol uptake rate oscillated with the same period as the biomass oscillations. These results show a dramatic increase in the ammonium and oxygen consumption rates prior to the initial budding of the synchronized subpopulation and a decrease in these rates during the late budding phase. At a pH of 5.5, the late budding phase is characterized by high specific ethanol productivity; however, the ethanol productivity lags the late budding phase at a pH pf 6.5. The observed time-varying metabolism in the oscillatory operating regime appears to be the result of the metabolic changes which occur during the cell cycle. Models which can predict the oscillatory biomass concentration and nutrient levels in this regime must be capable of predicting the concentrations and metabolic rates of the subpopulations as well.
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  • 18
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    Biotechnology and Bioengineering 36 (1990), S. 39-46 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Mandelonitrile lyase (EC 4.1.2.10) catalyzes the formation of D-mandelonitrile from HCN and benzaldehyde. Mandelonitrile lyase was immobilized by adsorption to support materials, for example, Celite. The enzyme preparations were used in diisopropyl ether for production of D-mandelonitrile. In order to obtain optically pure D-mandelonitrile it was necessary to use reaction conditions which favor the enzymatic reaction and suppress the competing spontaneous reaction, which yields a racemic mixture of D, L-mandelonitrile. The effects of substrate concentrations, water content, and support materials on both the spontaneous and enzymatic reactions were studied. The enzymatic reaction was carried out under conditions where the importance of the spontaneous reaction was negligible and high enantiomeric purity of D-mandelonitrile was achieved (at least 98% enantiomeric excess). The operational stability of the enzyme preparations was studied in batch as well as in continuous systems. It was vital to control the water content in the system to maintain an active preparation. In a packed bed reactor the enzyme preparations were shown to be active and stable. The reactors were run for 50 h with only a small decrease in product yield.
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  • 19
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    Biotechnology and Bioengineering 36 (1990), S. 55-63 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The cyclic, semicontinuous production of L-phenylacetyl carbinol (L-PAC) from a benzaldehyde substrate by Saccharomyces cerevisiae ATCC 834 immobilized in calcium alginate beads was substantially enhanced to about 4.5 g/L in a second cycle by reactivation in fresh medium for 24 h, following an earlier 24-h period of production from substrate. Intermittent feeding of benzaldehyde was employed (four doses in 3 h). In subsequent similar cycles, however, the production returned to that produced in the first cycle, viz. L-PAC concentration of 2-3 g/L in the medium. Production of L-PAC was also increased by adaptation of the cells over 200 h of exposure to the benzaldehyde substrate (compared to wild-type cells) and by continuous (as compared to intermittent) feeding of the substrate. A liter as great as 10 g/L was obtained with wild-type cells by continuous feeding of benzaldehyde over 6 h. Immobilization not only protected the cells from toxic effects of substrate but also permitted them to be used during 7 cycles of semicontinuous operation over more than 200 h.
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  • 20
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    Biotechnology and Bioengineering 36 (1990) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 21
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    Biotechnology and Bioengineering 36 (1990), S. 142-148 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A bioenergetic model has been developed for the fermentation of glucose by Bacillus polymyxa. This model uses energy balances to determine which pathways are utilized by the substrate. The model can predict substrate consumption, biomass formation, and the product distribution for this fermentation. The products are carbon dioxide, water, 2,3-butanediol, and ethanol, where ethanol represents lumped anaerobic products.
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  • 22
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    Biotechnology and Bioengineering 36 (1990), S. 166-178 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The notion that the regulated and flux-controlling enzyme in a metabolic network need not correspond suggests that the purpose of regulation may not be flux homeostasis under all physiological circumstances. Additionally, the fact that diversity in the function of intact metabolic networks exists suggests that in addition to time constant separation, other kinetic structure/regulatory mechanism patterns exist. In order to compliment and expand prior work on identifying kinetic structure-property relationships in networks, the present work explores in a general way how the control, dynamic, and energetic properties of metabolic networks depend on operating point, kinetic structure, and regulatory mechanism. The basic feature of trade-offs between properties is illustrated and used as a basis for indicating how particular subsets of structure, regulatory mechanism, and operating point emphasize certain properties that can be associated with a physiological function. Examples of scavenging trace metabolites and amphibolite coordination are proposed. Microstructure logic in terms of turnover number distributions as well as a potential mixed polynomial network analysis approach are also discussed.
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  • 23
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    Biotechnology and Bioengineering 36 (1990), S. 198-206 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The energy of the total transmitted light was subtracted from that of the incident light in a culture vessel and the difference was divided by the weight of cells. The value thus obtained was defined as the amount, Ex, of light energy absorbed per unit cell weight per unit time.Batch and continuous cultures of Chlorella vulgaris were carried out at 30°C in the pH range of 6.4-6.7 while restricting illumination. Next the specific growth rate, μ, in the batch culture and the fixed dilution rate, D, in the continuous culture were plotted against Ex. The results showed that the relation between D and Ex can be expressed in a Michaelis-Menten equation, where the maximal specific growth rate is 0.24 h -1 and the saturation constant is 6.58 kcal/g · h.Cell concentration calculated by substituting the apparent concentration, Xe, of incubated cells and the apparent maintenance constant, Me, for this equation agreed with that observed in almost all growth phases. Furthermore, from the change of chlorophyll productivity and the relationship between D and Ex expressed in this equation, it is assumed that Ex involves the light energy directly utilized in photosynthesis in the cells and that which is converted into, e.g., heat. This equation also indicated that a maximum in the growth yield existed. Then the growth yield of 0.029 g/kcal obtained at the incident light of 1.46 or 2.63 cal/cm2 · h was maximum (maximal conversion efficiency of light energy, 15.6%).These results indicate that this method of deriving the equation for the growth rate from this study is a useful procedure for obtaining bioengineering findings.
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  • 24
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    Biotechnology and Bioengineering 36 (1990), S. 233-242 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Accurate estimates of plasmid copy number in a cell are a prerequisite for predicting plasmid stability and protein production. A refined version of a structured model for the pBR322 plasmid replication mechanism is described. The model is capable of accurately predicting pBR322 plasmid copy number in Escherichia coli B/r for a wide range of growth rates. The refinements include better estimates of promoter strength, the degradation rate of RNA species, binding constant of RNAI-RNAII reaction, and dependency of promoter strength on growth rate. The predictions of the model are verified by recent experimental observations but differ from some previous reports. This model can also be used to predict the binding constant of the RNAI-RNAII reaction of ColE1 type plasmids. At 37°C, the binding constant is estimated to be 77 ± 11 × 10-13 mL/molecule-h for pBR322.
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  • 25
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    Biotechnology and Bioengineering 35 (1990), S. 691-701 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Predictions may be made for the influence of solvent choice on the equilibrium position of biocatalyzed reactions, based on data for the liquid-liquid distribution of the reactants. The most reliable predictions are probably for dilute systems, based on partition coefficients or correlations derived from them. The effective equilibrium constant for esterification reactions is predicted to alter by more than four orders of magnitude on changing between different water-immiscible solvents. The equilibrium constant correlates well with the solubility of water in the solvent, and is most favorable for synthesis in the least polar solvents (aliphatic hydrocarbons). Similar effects seem to apply for other reactions, including oxidation of alcohols and hydrolysis of chlorides. Predictions can be made for nondilute systems using the UNIFAC system of group contributions, but the reliability of these is more questionable.
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  • 26
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    Biotechnology and Bioengineering 35 (1990), S. 727-731 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pichia stipitis NRRL Y-7124 is a xylose-fermenting yeast able to accumulate ca. 57 g/L ethanol. Because optimum process conditions are important, data were collected to determine the effects of temperature and pH on growth and fermentation rates and product accumulations. Temperatures (26-35°C) providing optimum biomass and ethanol productivities did not necessarily provide maximum ethanol accumulation. Xylitol and residual xylose concentrations increased with temperature. Maximum ethanol selectivity was achieved at 25-26°C with minimal sacrifice to production rates. The temperature optimum for xylose could not be generalized to glucose fermentations, in which ethanol productivity and accumulation were optimum at 34°C. The optimum pH range for growth and fermentation on xylose was 4-7 at 25°C.
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  • 27
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Production of various extracellular enzymes (the β-lactamases from Streptomyces albus G, Streptomyces cacaoi, Actinomadura R39, and the DD-carboxypeptidase from Streptomyces R61) by genetically engineered Streptomyces lividans TK24 in Lennox broth medium reached a maximum after 36 to 48 h. Subsequently, the enzyme activity drastically decreased probably due to an increased pH value and the production of an inactivator by Streptomyces lividans. Protease activity did not seem to play a major role. The increased pH and inactivator synthesis are related to amino acid catabolism and generally result in cellularlysis. The use of a medium where the catabolism of amino acids was made less likely by the presence of glucose and NH4Cl and by buffering at pH 7.4 considerably inproved the yield. Furthermore, the water activity of the medium seemed to be an important parameter for the production of extracellular proteins by genetically engineered Streptomyces. Better production was observed when the water activity was decreased to 0.96-0.98 by addition of sucrose.Under those conditions, the concentration of extracellular enzyme reached about 0.3 g (1 g in the best case)/L of culture supernantant.
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    Biotechnology and Bioengineering 36 (1990), S. 207-217 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Acetone-butanol-ethanol (ABE) fermentation was performed continuously in an immobilized cell, trickle bed reactor for 54 days without, degeneration by maintaining the pH above 4.3. Column clogging was minimized by structured packing of immobilization matrix. The reactor contained two serial glass columns packed with Clostridium acetobutylicum adsorbed on 12- and 20-in.-long polyester sponge strips at total flow rates between 38 and 98.7 mL/h. Cells were initially grown at 20 g/L glucose resulting in low butanol (1.15 g/L) production encouraging cell growth. After the initial cell growth phase a higher glucose concentration (38.7 g/L) improved solvent yield from 13.2 to 24.1 wt%, and butanol production rate was the best. Further improvement in solvent yield and butanol production rate was not observed with 60 g/L of glucose. However, when the fresh nutrient supply was limited to only the first column, solvent yield increased to 27.3 wt% and butanol selectivity was improved to 0.592 as compared to 0.541 when fresh feed was fed to both columns. The highest butanol concentration of 5.2 g/L occurred at 55% conversion of the feed with 60 g/L glucose. Liquid product yield of immobilized cells approached the theoretical value reported in the literature. Glucose and product concentration profiles along the column showed that the columns can be divided into production and inhibition regions. The length of each zone was dependent upon the feed glucose concentration and feed pattern. Unlike batch fermentation, there was no clear distinction between acid and solvent production regions. The pH dropped, from 6.18-6.43 to 4.50-4.90 in the first inch of the reactor. The pH dropped further to 4.36-4.65 by the exit of the column. The results indicate that the strategy for long term stable operation with high solvent yield requires a structured packing of biologically stable porous matrix such as polyester sponge, a pH maintenance above 4.3, glucose concentrations up to 60 g/L and nutrient supply only to the inlet of the reactor.
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  • 29
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    Biotechnology and Bioengineering 36 (1990), S. 224-232 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: If a microorganism has a growth coupled production or consumption of acid or alkali, it is possible to use the pH-auxostat as a means of control in continuous fermentation. In using the pH-auxostat, it is possible to separate the inlet substrate flow in two different streams. These will both be pH controlled, with one main flow, consisting of nutrients and a second minor but concentrated flow, of acid or alkali. Hereby, it is possible to vary the difference in pH between the fermentor and the inlet medium. This pH difference is proportional to the steady-state cell mass concentration.1,2 It is shown that by separating the inlet flow in two different streams and cultivating without any substrate limitation, the maximum growth rate may be obtained while the cell mass concentration will be controlled. This will also give the possibility to reach high cell mass concentrations at μmax without the risk of wash-out. A modified expression, based on hydrogen, of the steady-state bio-mass concentration, X, is developed as \documentclass{article}\pagestyle{empty}\begin{document}$$ X = Y_{X/H} \cdot [F_{{\rm Hin}} /(F_{{\rm Hin}} + F_{{\rm Min}} )] \cdot (C_{{\rm Hin}} - C_{{\rm HFERM}} ) $$\end{document} where YX/H is the yield coefficient of cell mass per acid produced. The indexes Hin and Min refer to the inflows of alkali and medium, respectively; CHin is the inlet concentration of hydrogen ions. The boundary condition for the cell mass shows that Sin 〉 X/YX/S, where Sin is the medium substrate concentration and YX/S is the yield of biomass per consumed substrate. It is shown that when the cell mass concentration exceeds this value, the flow stops. The applicability of the pH-auxostat method is then verified from different experiments. It is hereby used to detect a deviation from the maximal growth rate showing effects on the microbial physiology. With Escherichia coli used as the model organism, the effect on the growth rate of temperature and high concentration of ammonia were investigated.
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  • 30
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    Biotechnology and Bioengineering 36 (1990), S. 263-269 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We prepared artificial cells each containing leucine dehydrogenase (EC 1.4.1.9), urease (EC 3.5.1.5), soluble dextran-NAD+, and one of the following coenzyme regenerating dehydrogenases: glucose dehydrogenase (EC 1.1.1.47); yeast alcohol dehydrogenase (EC 1.1.1.1); malate dehydrogenase (EC 1.1.1.37); or lactate dehydrogenase (EC 1.1.1.27). Artificial cells were packed in small columns. L-Leucine, L-valine, and L-isoleucine were continuously produced with simultaneous dextran-NADH regeneration. The maximum production ratios depended on the coenzyme regenerating systems used: 83-93% for D-glucose and glucose dehydrogenase system; 90% for ethanol and yeast alcohol dehydrogenase system; 45-55% for L-malate and malate dehydrogenase system; and 64-78% for L-lactate and lactate dehydrogenase system. Kinetic experiments were also carried out. The apparent Km values are as follows: 0.33 mM for α-ketoisocaproate (KIC); 0.51 mM for α-ketoisovalerate (KIV); 0.58 mM for DL-α-keto-β-methyl-n-valerate (KMV); 3.52 mM for urea; 27.82 mM for D-glucose; 3.89 mM for ethanol; 3.02 mM for L-malate; and 16.67 mM for L-lactate. Kinetic analysis showed that KIC, KIV, and KMV were all competitive inhibitors in the reactions catalyzed by leucine dehydrogenase. Their inhibitor constants were the corresponding Km values.
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  • 31
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    Biotechnology and Bioengineering 36 (1990), S. 1070-1082 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A method is presented for the estimation of the standard Gibbs energies of formation of biochemical compounds (and hence the Gibbs energies and equilibrium constants of biochemical reactions) from the contributions of groups. The method employs a large set of groups and special corrections. The contributions were estimated via multiple linear regression, using screened and weighted literature data. For most of the data employed, the error is less than 2 kcal/mol. The method provides a useful first approximation to Gibbs energies and equilibrium constants in biochemical systems.
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  • 32
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    Biotechnology and Bioengineering 36 (1990), S. 1105-1109 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: It was observed that about 90% of free-swimming Thiobacillus ferrooxidans in 9 K medium was adsorbed on added activated carbon when the concentration of the cultivated bacteria reached about 4 × 1013 cells m-3. The oxidation of ferrous iron and the leaching of copper ore were carried out in shake flasks and in aerated columns. The rates of oxidation and leaching increased when bacteria adsorbed on activated carbon were used. However, the evaluation of the reaction rates by eliminating the catalytic effect of activated carbon showed that the contribution to the reaction by the adsorbed microorganism was very small.
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  • 33
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    Biotechnology and Bioengineering 36 (1990), S. 1133-1140 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In order to develop an efficient process for large-scale production of recombinant protein, various factors were studied which affect the productivity of Sf-9 (Spodoptera frugiperda) insect cells when using the baculovirus expression system. It was shown that upon infection with the Bac-BRV6L recombinant baculovirus, the level per cell of VP6 (a bovine rotavirus nucleocapsid protein) would drop 10-fold when host cell density at the time of infection increased from 2 × 106 to 3 × 106 cells/mL. The decrease was found to be totally reversible by culture medium renewal after infection, even when cells were infected at the stationary phase. Recombinant protein production was 4-6 times higher using TNMFH medium supplemented with 10% fetal bovine serum (FBS) than in IPL/41 serum-free medium. Fine-tuning of infection parameters in a 4-L surface-aerated bioreactor resulted in the production of typically 350 mg/L of VP6 protein, representing more than 25% of total cell proteins.
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  • 34
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    Biotechnology and Bioengineering 37 (1991) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 35
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    Biotechnology and Bioengineering 37 (1991), S. 17-25 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Bacterial biofilm removal processes due to shear and catastrophic sloughing have been investigated in a turbulent flow system under conditions of carbon versus oxygen substrate limitations and varying aqueous phase calcium concentrations. Biofilm cellular and extracellular polymer carbon, total biofilm carbon and mass, and biofilm calcium concentrations are measured for pure culture biofilms of the facultative aerobe, Pseudomonas putida ATCC 11172. Results indicate oxygen-limited biofilms reach a higher steady-state biofilm organic carbon level than carbon-limited biofilms. Oxygen-limited biofilms also exhibit (1) a higher extracellular polymer-carbon: cell-carbon ratio throughout biofilm development and (2) a higher biofilm calcium content than carbon-limited biofilms. Increasing aqueous phase calcium concentrations increase the amount of biofilm calcium in both cases; the rate of calcium accumulation in oxygen-limited biofilms increases with increasing liquid phase calcium concentrations over the entire range studied while the rates of calcium accumulation in carbon-limited biofilms appear independent of aqueous phase calcium concentrations above 11.0 mg/L. Oxygen-limited biofilms with their higher extracellular polymer and calcium content exhibit shear removal rates that are 20-40% of those observed for carbon-limited biofilms. However, it is the oxygen-limited biofilms that experience catastrophic sloughing events. The carbon-limited biofilms studied here never sloughed even if subjected to intentional long-term deprivation of all nutrients. Reduced shear removal and the susceptibility to sloughing of the oxygen-limited biofilms are attributed to their more cohesive structure bought about by their relatively greater extracellular polymer production.
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  • 36
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The growth of two lymphocyte cell lines, a hybridoma cell line and a human cutaneous T cell lymphoma (HuT78), was studied in fed-batch culture, and unstructured models of growth developed. A criteria was established to insure that the growth rate varied by less than a specified tolerance throughout the culture period. Glutamine and serum were growth-limiting nutrients for both cell lines with half-maximal growth rates at 0. 53 mM glutamine and 0. 55%(v/v) serum for the hybridoma cells and 0. 21 mM glutamine and 1. 5% serum for the HuT-78 cells. Over the range of glucose concentrations from 5. 5 mM to 28 mM, the specific growth rate of hybridoma cells was independent of glucose concentration, whereas glucose concentrations above 5. 5 mM inhibited HuT-78 growth. For both cell lines, the growth rate was significantly inhibited by the addition of ammonium, although the hybridoma cell line was more affected by ammonia than was the HuT-78 cell line. Growth of HuT-78 cells increased in the presence of interleukin-2. Unstructured models for the hybridoma cells were similar to other models presented in the literature. Applications of these models to adoptive immunotherapy are discussed.
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  • 37
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    Biotechnology and Bioengineering 36 (1990), S. 547-562 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A stochastic model is proposed to simulate the growth of anchorage dependent cells on a flat surface. The model, based on representing the cell shapes on the surface as external irregular polygons with the nuclei distributed as a set of Poisson points (producing a modified Voronoi tessellation of 2 space) and incorporating a distribution function to describe cell division of the perimeter cells of the colony, provides data not only on population dynamics but also on the patterns produced by clusters of cells in the colony. These patterns produced by the model are qualitatively similar to observations reported for some cell cultures. The periods of induction, rapid growth, and decreasing growth asymptoting to zero as confluence is reached are predicted by the model. Quantitative comparison with published experimental data for this is good. The specific growth rate computed for the period of rapid growth predicted by the model is dependent on the distribution function describing the cell division time. As the standard deviation of this increases, the specific growth rate decreases as with a consequent increase in time to achieve confluence. The removal of cells from the colony by shear forces or death is considered in the model. As the probability for removal increases, the cell density at confluence and specific growth rate decrease. The clusters of cells, patterns, in the colony are very sensitive to cell removal. By analyzing these patterns in experiments, an estimate of cell removal can be made. The areas covered by cells on a substrate are fractal patterns. The fractal dimension is always greater than 1 and is a function of the removal probability.
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  • 38
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    Biotechnology and Bioengineering 36 (1990), S. 593-600 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A lipoprotein lipase (LPL) was made water insoluble by immobilizing onto the surface of polyacrolein (PAA) microspheres with and without oligoglycines as spacer. The activity of the immobilized LPL was found to remain high toward a small ester substrate, p-nitrophenyl laurate (pNPL). The relative activity of the immobilized LPL without spacer decreased gradually with the decreasing surface concentration of the immobilized LPL on the PAA microsphere. On the contrary, the immobilized LPL with oligoglycine spacers gave an almost constant activity for the substrate hydrolysis within the surface concentration region studied and gave a much higher relative activity than that without any spacer. The Michaelis constant Km and the maximum reaction velocity Vm were estimated for the free and the immobilized LPL. The apparent Km was larger for the immobilized LPL than for the free one, while Vm was smaller for the immobilized LPL. The pH, thermal, and storage stabilities of the immobilized LPL were higher than those of the free one. The initial enzymatic activity of the immobilized LPL maintained almost unchanged without any leakage and inactivation of LPL when the batch enzyme reaction was performed repeatedly, indicating the excellent durability of the immobilized LPL.
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  • 39
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    Biotechnology and Bioengineering 36 (1990), S. 630-635 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of cell density as well as the concentration levels of glucose and glutamine on the specific respiration rate of a hybridoma cell line were investigated. The experimental oxygen consumption rate was found to be constant over a wide range of dissolved oxygen levels if the suspension medium contained glutamine. In glutamine-free medium, however, the rate of oxygen consumption decreased slowly with time.In a stationary flask batch culture, the specific respiration rate decreased from about 7 to 2.9 μmol/min per 109 cells as the cell density increased exponentially from 1 × 105 to 1.2 × 106/mL. To isolate the effect of cell density, cells were re suspended in fresh culture medium so that nutrient concentrations were the same for all experiments. The specific respiration rate decreased with increasing cell density in the same manner as in the stationary flask culture, falling from 8 to 4 μmol/min per 109 cells as the cell density increased from 105 to 106 cells/mL, then declining to 2 μmol/min per 109 cells when the cell density reached 107 cells/mL.Cells suspended in Hanks balanced sale solution (HBSS) were used to elucidate the effect of glucose and glutamine levels on respiration. The addition of glucose in concentrations of 0.25, 0.50, and 0.75 g/L had no observable effect on the specific oxygen uptake rate; however, a glucose concentration of 1 g/L reduced the uptake rate by 22%. Glutamine in a concentration of 0.30 g/L increased the specific respiration rate in HBSS containing 0 and 1 g/L glucose by approximately 13%.
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  • 40
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    Biotechnology and Bioengineering 36 (1990), S. 970-973 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 41
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    Biotechnology and Bioengineering 36 (1990), S. 983-992 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The viable fraction of immobilized cells in a bioreactor may be critical in predicting long-term or steady-state reactor performance. The assumption of near 100% viable cells in a bioreactor may not be valid for portions of immobilized cell reactors (ICRs) characterized by conditions resulting in appreciable death rates. A mathematical model of an adsorbed cell type ICR is presented in which a steady-state viable cell fraction is predicted, based on the assumptions of no cell accumulation in the reactor and a random loss of cells from the reactor. Data on cell death rates, cell growth rates, and productivity rates as functions of temperature, substrate, and ethanol concentration for the lactose utilizing yeast K. fragillis were incorporated into this model. The steady-state reactor viable cell fraction as predicted by this model is a strong function of both temperature and ethanol concentration. For example, a stable 20% viable fraction of the immobilized cells is predicted in ICR locations experiencing continuous conditions of either 30 g/L ethanol at 40°C, or 95 g/L ethanol at 25°C. Steady-state ICR “plug flow” concentration profiles and column productivities are predicted at three operating temperatures, 20, 30, and 40°C using two different models for ethanol inhibition of productivity. These profiles suggest that the reactor operating temperature should be low if higher outlet ethanol concentrations are desired. Three reactor design strategies are presented to maximize the viable cell fraction and improve long-term ethanol productivity in ICR's: (1) reducing outlet ethanol concentrations, (2) rotating segments of an ICR between high and low ethanol environments, and (3) simultaneous removal of the ethanol produced from the reactor as it is formed.
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  • 42
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The growth and metabolism of Saccharomyces cerevisiae was studied in steady-state chemostat cultures under conditions of scarce oxygen and excess glucose. The specific ethanol productivity and specific glucose uptake rate were stimulated by 50% within a narrow range of air/nitrogen mixtures to the fermentor. Fermentation was inhibited at slightly higher and lower air/nitrogen ratios, confirming similar results by previous investigators. This stimulation could not be caused by obvious mechanisms, such as the Pasteur or Crabtree effects. Since this maximum in the fermentation rate occurred in a steady-state chemostat and at a constant dilution rate, the ATP yield of the culture necessarily attained a minimum. Thus, changes in the energetic efficiency of growth or the degree of wasting of ATP were surmised. The steady-state biomass concentration at various oxygenation rates exhibited hysteresis phenomena. Ignition and extinction of the biomass concentration occurred as critical oxygen feed rates were passed. The hysteresis was prevented by adding yeast extract to or reducing the antifoam concentration in the medium. These medium alterations had the simultaneous effect of stimulating the fermentation rate, suggesting that ATP has a critical role in dictating the biomass concentration in micro-aerobic culture. Silicone polymer antifoam was found to stimulate glycerol production at the expense of ethanol production, having consequences for the energy generation and the biomass concentration of the culture.
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  • 43
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    Biotechnology and Bioengineering 36 (1990), S. 1049-1055 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Hybridoma cells (S3H5/γ2bA2) were cultivated in spinner flasks with 1% serum media and serum-free media. Monoclonal antibody productivity was maintained in 1% serum media. However, cells in serum-free media showed a decrease in antibody productivity, and it completely disappeared in IMDM-based low protein medium. This loss of antibody productivity was not observed when the cells were immobilized in alginate beads. In fact, immobilization enhanced the specific MAb productivity.
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  • 44
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    Biotechnology and Bioengineering 37 (1991), S. 491-496 
    ISSN: 0006-3592
    Keywords: potentiostatic control of enzyme activity ; immobilized enzyme ; graphite ; conducting support ; specific anion inhibition ; carbonic anhydrase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of both a positive and a negative applied potential on the p-NPA hydrolysis activity of bovine carbonic anhydrase (BCA) immobilized on graphite rods has been investigated. Background experiments show that the pH-activity profile for BCA free in solution is not affected by either a negative or a positive potential applied to graphite rods placed in the same solution. However, the activity of BCA immobilized by covalent attachment to a graphite rod is influenced by a potential externally applied to the graphite rod. An overall increase in activity (as determined by the initial rate of the p-NPA hydrolysis reaction) is observed in the presence of a -0.2 V (Ag/AgCl) applied potential, while decreased activity is evident at +0.6 V (Ag/AgCl). This is indicative of an electrolyte anion effect rather than a local pH effect. In the presence of the specific anion inhibitors Cl- and SCN-, the relative BCA activity increases at -0.2 V (Ag/AgCl) and decreases at +0.6 V (Ag/AgCl) are consistent with the different BCA inhibition constants for Cl- and SCN-. Accelerated loss of immobilized BCA activity also accompanies the application of the external potentials, particularly at +0.6 V (Ag/AgCl). Results described here represent an early example of potentiostatic control of nonredox enzyme activity. Several possible mechanisms are discussed including specific anion inhibition, enzyme surface charge/charged support material interactions, and charged product inhibition. It is likely that a combination of such mechanisms is operational in this system. The implications of external potentials affecting the activity of immobilized enzymes in the design of stable immobilized enzyme electrodes are also discussed.
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  • 45
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    Biotechnology and Bioengineering 37 (1991), S. 528-536 
    ISSN: 0006-3592
    Keywords: yeast thermotolerance ; Saccharomyces ; invertase ; sugarcane juice fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Maintenance of high cell viability was the main characteristic of our new strains of thermotolerant Saccharomyces. Total sugar conversion to ethanol was observed for sugarcane juice fermentation at 38-40°C in less than 10 h and without continuous aeration of the culture. Invertase activity differed among the selected strains and increased during fermentation but was not dependent on cell viability. Invertase activity of the cells and optimum temperature for growth, as well as velocity of ethanol formation, were dependent on medium composition and the type of strain used. At high sugarcane syrup concentrations, the best temperature for ethanol formation by strain 781 was 35°C. Distinct differences among the velocities of ethanol production using selected strains were also observed in sugarcane syrup at 35-38°C.
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  • 46
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    Biotechnology and Bioengineering 35 (1990), S. 132-137 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Six different types of materials including PVC, chitosan, chitin, agarose, Sepharose, and Trisacryl were evaluated for their lipase-coupling efficiencies. Among those tested, chitosan yielded the highest amount of lipase (79 mg/mL packed gel) immobilized but with lowest oil hydrolytic activity (0.03 mg eq/mL gel). The amount of lipase immobilized was affected by the length of the hydrocarbon chain attached to the PVC matrix but not by the pore size of the supports used. On the other hand, the specific activity of the immobilized lipase was affected by the pore size but not by the chain length of the hydrocarbon attached to the support. After immobilization, the optimal reaction pH was shifted from 7.5 to 8.5 and the optimal reaction temperature from 35 to 45-55°C. Lipase immobilized on PVC exhibited higher thermal stability than that on agarose. The half-life of the PVC immobilized lipase operating at 30°C in a packed-bed reactor was estimated to be about 400 h.
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  • 47
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    Biotechnology and Bioengineering 36 (1990), S. 834-838 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Alcohol oxidase, an enzyme which exhibits relatively weak substrate specificity among short chain alcohols, forms the corresponding aldehyde and hydrogen peroxide as coproduct. The ability of alcohol oxidase from Pichia pastoris yeast to convert ethanol to acetaldehyde and hydrogen peroxide was examined in an oxygen pressure reactor under conditions, such that oxygen availability was sufficient to permit rapid catalysis. Hydrogen peroxide levels of ∼1.8/M (6% w/w) were attained in 2-3 h with 2.8 μM enzyme, corresponding to a productivity of ∼30 g peroxide/g enzyme. Optimal conditions (within equipment limitations) were 900 psi oxygen, 2.6M ethanol, at 4 °C. Similar levels of products were reached in the reactor using enzyme immobilized covalently on controlled pore glass and noncovalently on an anion exchange support. Recycle of covalently immobilized enzyme was not possible as a result of enzyme inactivation after a single run. Limited recycle of noncovalently immobilized enzyme was accomplished with substantial decreases in levels of product attainable on each cycle.
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  • 48
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    Biotechnology and Bioengineering 36 (1990), S. 854-864 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Even when microorganisms are grown in highly agitated fermentors, calculation predicts a mismatch between the microscale of the turbulence (where the smallest eddy is typically 50-300 μm diameter) and the cellular dimensions (1-5 μm). The cell thus spends substantial portions of time in an apparently stagnant eddy, depleted of nutrients. The local fluid microscales were measured in a laboratory fermenter to confirm this. Using S. cerevisiae in continuous culture, it is shown that the local microscales influence cell metabolism dramatically. The issues addressed in this study are thus micromixing and microsegregation of reactants and how they influence cell yield.
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  • 49
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    Biotechnology and Bioengineering 36 (1990), S. 887-901 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Escherichia coli cells were immobilized and grown in hollow-fiber reactors allowing simultaneous NMR spectroscopy and perfusion with nutrient medium. The extent to which the cells were starved due to inadequate mass transfer was predicted using a mathematical model of reaction and diffusion. Reactors were experimentally characterized using 35S autoradiography to visualize spatial variations in protein synthesis rates and transmission electron microscopy to indicate spatial variations in cell morphology. Mass transfer limitations in reactors operated at 37 °C were shown to be severe, with regions of starved cells occupying up to 80% of the cell-containing region. Phosphorus-31 nuclear magnetic resonance (NMR) spectra of the immobilized, perfused cells revealed abnormally low volume-averaged concentrations of sugar phosphates, NTP, and ratios of NTP/NDP in these reactors. Intracellular pH was also depressed in the cells. In order to overcome mass transfer limitations in the cell layer, the reactor growth temperature was decreased. Sulfur-35 autoradiographs of a reactor operated at 16°C did not indicate the presence of starved cells. The NMR spectra obtained from this reactor showed near-normal intracellular pH, metabolite concentrations, and NTP/NDP ratios. The presence of significant mass transfer limitations in a perfused cell sample during NMR spectroscopy is generally undesirable since the resulting spectra can be ambiguous and difficult to interpret. The strategy adopted in this work, namely estimation of the relative rates of reaction and diffusion in the cell mass and appropriate changes in reactor design and operating parameters, should prove generally applicable for the design of perfused cell samples for NMR spectroscopic experiments.
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    Biotechnology and Bioengineering 37 (1991) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 51
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    Biotechnology and Bioengineering 37 (1991), S. 716-722 
    ISSN: 0006-3592
    Keywords: κ-carrageenan matrix ; Lactobacillus delbrueckii ; toxicity ; solvent diffusion ; extractive fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The toxicity of an Alamine 336/oleyl-alcohol extraction system on Lactobacillus delbrueckii was investigated. It was shown that the solvent affected the cells through the water-soluble portion and the immiscible portion of the solvent. While immobilization significantly protected the cells from the immiscible solvent phase, the water-soluble part of the solvent still caused toxicity to the microorganisms due to diffusion of the solvent into the matrix. Adding soybean oil to the κ-carrageenan matrix could trap the diffusing solvent molecules, and therefore reduce the toxic effect from the water soluble portion of the solvent. The protective ability of soybean oil was quantified through mathematical modeling and experimentation.
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  • 52
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    Biotechnology and Bioengineering 37 (1991), S. 746-754 
    ISSN: 0006-3592
    Keywords: Lactococcus cremoris ; cell-recycle fermentor ; cross-flow filtration ; high cell concentration cultures ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: High-cell concentration cultivation of Lactococcus cremoris, a homofermentative lactic acid producer, in a cell-recycle fermentor is described. Cross-flow filtration allowing continuous removal of the inhibitory metabollte, the influence of dilution rate on growth was investigated in total or partial cell-recycle cultures. The dependence of growth characteristics on operating conditions was identified and quantified using lactose as the carbon source. Growth kinetics could be described by both lactate removal efficiency and nutrient availability. Based on physiological observations, biomass and lactic acid productivities were predicted in partial cell-recycle cultures.
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  • 53
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    Biotechnology and Bioengineering 37 (1991), S. 778-789 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Biofilm research has focused on studies of undefined mixed microbial populations and, more recently, on investigations of monopopulation biofilms. In the first case, the biofilm is considered a homogeneous mass, ignoring the properties of individual species. The second case concentrates on the properties and processes of one microbial species in the biofilm. This article describes biofilm experiments conducted with monopopulations of Klebsiella pneumoniae and Pseudomonas aeruginosa and with binary populations of K. pneumoniae and P. aeruginosa. Process rates and stoichiometric coefficients were determined for the monopopulation and for the binary population biofilms and evaluated in light of the species distribution in the latter. Results indicate that neither the specific cellular product formation rate nor the glucose-oxygen stoichiometric ratio of K. pneumoniae or P. aeruginosa in the binary biofilm is affected by the presence of the other species. Consequently, species interaction was not observed. Although the specific cellular growth rate of K. pneumoniae is five times that of P. aeruginosa, the former species did not dominate the microbial population in the biofilm. Possible reasons for this unexpected behavior are discussed.
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  • 54
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    Biotechnology and Bioengineering 37 (1991), S. 802-808 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simple structured model is described and compared with experimental data for fermentations with recombinant. Escherichia coli. The model is a so-called compartment, model, where the different biomass components are lumped together in a few intracellular variables. The model is able to describe, in a biologically reasonable fashion, a majority of the observations that have been made through fermentations with recombinant microorganisms. The model is especially suited for description of dynamic changes in plasmid copy number, e.g., runaway plasmid replication.
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  • 55
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    Biotechnology and Bioengineering 37 (1991), S. 834-842 
    ISSN: 0006-3592
    Keywords: microalgae ; bioreactor ; CO2 ; absorption ; KLa ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: For the characterization of CO2 absorption in aerated microalgal culture systems, a different approach based on KLa(O2) determination and transformation was studied. To confirm the validity of this method, the influence of reactions between CO2 and compounds (OH-, H2O, and NH3) present in the culture medium upon the absorption mechanism was evaluated under different physical and chemical culture conditions. Under these conditions, knowledge of the relative magnitudes of the diffusion and reaction kinetics permitted the evaluation of their relative importance. For the determination of the parameters required for the calculation of the CO2 absorption constant, empirical correlations for KL0 and a were used that had been previously verified with experimental data for O2 absorption. Since, for the conditions studied, the absorption rate was shown to be independent of the chemical reactions taking place in the liquid phase, the KLa for CO2 could be directly related to the KLa for O2 by a simple factor that took into account the difference in aqueous diffusivity of the two gases. Thus, using methods developed for determining O2 absorption in gas-liquid contactors, it is possible to adequately characterize CO2 absorption for laboratory and pilot scale algal production systems.
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    Biotechnology and Bioengineering 37 (1991), S. 883-888 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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    Biotechnology and Bioengineering 37 (1991) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 37 (1991), S. 918-921 
    ISSN: 0006-3592
    Keywords: plant cells ; photosynthesis ; microemulsion ; organic solvents ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: With the aim of possibly extending plant microbiology and photosynthesis beyond the usual applicability in aqueous solution, we investigated the solubilization of plant cells inorganic media with the help of water-in-oil microemulsions. Cells isolated from leaves of Rumex obtusifolius were solubilized in a water/2-ethyl-hexyl-sodiumsulfosuccinate/isooctane system, containing 20% water (v:v) and 240 mM surfactant, and the oxygen evolution/consumption was measured polarographically. Although no oxygen evolution was detectable in the organic medium, the cells were able to carry out photosynthetic oxygen consumption at the expense of ascorbate. To a lesser extent, photosynthetic oxygen consumption was measured using N, N, N′, N′-tetramethyl-p-phenylenediamine as electron donor. The rate of ascorbate photooxidation was linearly related to the concentration of cells.
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  • 59
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The marine phytoplankter Tetraselmis suecica was grown in shallow outdoor flumes for a period of approximately 6 months at the Natural Energy Laboratory of Hawaii. In full sunlight, gross production rates were 15-20 g C m-2 d-1. The corresponding photosynthetic efficiencies (PE's) were 9-10%. Respiration losses removed about half the gross production. The CO2 utilization efficiencies of 96 ± 11% were achieved by bubbling CO2 into the culture with the use of a counterflow sump system. Adding the CO2 in the form of carbonated water resulted in utilization efficiencies of 81 ± 11%. Archimedes screws proved superior to both paddle wheels and propellers as a means of circulating the water in the flumes. Insertion of foil arrays into the flumes to effect systematic mixing of the culture significantly enhanced production. The enhancement was greater when the foils were oriented at a small angle relative to the horizontal than when they were oriented at the same angle relative to the vertical. Light modulation effects are implicated as the probable cause of most of the enhancement. Substitution of electric power plant stack gases for pure CO2 resulted in no significant change in the production of T. suecica grown in chemostat culture.
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    Biotechnology and Bioengineering 37 (1991), S. 967-972 
    ISSN: 0006-3592
    Keywords: papain ; organic solvent ; kinetics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of various concentrations of a water-miscible organic solvent [a 7:3 (v/v) mixture of N, N dimethylformamide and dimethylsulfoxide] on the kinetics of papain have been investigated. The parameters kcat and Km for the amidase and esterase activity of papain using N-α-benzoyl-DL-arginine-p-nitroanilide (BAPNA) and N-α-benzoyl-L-arginine ethyl ester (BAEE) as substrates were determined. For both types of activity, kcat initially increased (up to about 15% solvent), and then decreased with increasing concentrations of organic solvent. In contrast, Km increased sharply with the organic solvent concentration. Active site titration at 0 and 50% solvent indicated no change in the amount of active enzyme. Fluorometric measurements of the emission spectrum of papain did not indicate any major conformational changes with increasing concentrations of organic solvent.
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    Biotechnology and Bioengineering 37 (1991), S. 383-385 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 62
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Notes: Ethanol and cycloheximide inhibited the function of the ammonium transport system in growing cultures of Saccharomyces cerevisiae var. ellipsoideus measured as methylamine uptake. The effect was reversible with ethanol and irreversible with the antibiotic. The kinetic data are consistent with a reduction of the number of active carrier molecules located in the plasma membrane. In contrast, neither ethanol nor cycloheximide affected the specific rate of fructose uptake.
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    Biotechnology and Bioengineering 37 (1991), S. 397-403 
    ISSN: 0006-3592
    Keywords: immobilized Plant cells ; plant cells bioreactor ; Protoberberine Alkaloids ; Thalictrum rugosum ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Cultured Thalictrum rugosum cells were immobilized using a glass fiber substratum previously shown to provide optimum immobilization efficiency based on spontaneous adhesion mechanisms. When cultivated in shake flasks, immobilized cells exhibited decreased growth and protoberberine alkaloid production rates in comparison to freely suspended cells. Since alkaloid production is growth associated in T. rugosum, the decreased specific production rate was a function of the slower growth rate. Cells immobilized on glass fiber mats appear to be amenable for extended culture periods. Maximum biomass and protoberberine alkaloid levels were maintained for at least 14 days in immobilized cultures. In contrast, fresh weight, dry weight, and total alkaloid content decreased in suspension cultures following the linear growth phase.Glass fiber mats were incorporated in to a 4.5-L plant cell bioreactor as horizontal disks supported on a central rod. Mixing in the reactor was provided by the combined actions of a magnetic impeller and a cylindrical sparging colum. fThe magnetic impeller and a cylindrical sparging column. The entire inoculum biomass of T. rougosum, introduced as suspension, was spontaneously immobilized with in 8h. During liner phase, the growth rate of bioreactor cultivated immobilized cells (μ = 0.06 day-1) was 50% that immobilized cell viability in both systems was determined to be similar. The increase in specific production of protoberberine alklodis was initially similar in bioreactor-and culture period. The increase in specific production of protoberberine alkaloids was initially similar in bioreactor-and shake-flask-cultivated immobilized cells. However, the maximum specific production of bioreactor grown cultures was lower. The scale up potential of an immobilization strategy based on the spontaneous adhesion of immobilization strategy based on the spontaneous adhesion of cultured plant cells to glass fiber is demonstrated.
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    Biotechnology and Bioengineering 37 (1991), S. 190-195 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstrct.
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    Biotechnology and Bioengineering 37 (1991), S. 205-209 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The oxygen transfer characteristics of a 20-mm O.D. airlift contactor fitted with an oxygen microelectrode were determined by steady-state sulfite oxidation measurements. The volumetric mass transfer coefficient kLa was proportional to sparging power input per unit volume raised to a power which varied from 0.41 in water (coalescing bubbles) to 0.76 in NaCl solutions (noncoalescing bubbles). The highest observed kLa value was 0.012 s-1 which is sufficient to aerate Escherichia coli in an NMR spectrometer at moderate to high cell densities, depending on the physiological state of the cells.
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    Biotechnology and Bioengineering 37 (1991), S. 467-483 
    ISSN: 0006-3592
    Keywords: alkaline protease ; Bacillus firmus ; extracellular enzymes ; nitrogen/oxygen/phosphorous limitation ; acetic acid ; ethanol ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Proteolytic enzymes produced by Bacillus species find a wide variety of applications in brewing, detergent, food, and leather industries. Owing to significant differences normally observed in culture conditions promoting cell growth and those promoting production of metabolites such as enzymes, for increased efficacy of bioreactor operations it is essential to identify these sets of conditions (including medium formulation). This study is focused on formulation of a semidefined medium that substantially enhances synthesis and secretion of an alkaline protease in batch cultures of Bacillus firmus NRS 783, a known superior producer of this enzyme. The series of experiments conducted to identify culture conditions that lead to improved protease production also enables investigation of the regulatory effects of important culture parameters including pH, dissolved oxygen, and concentrations of nitrogen and phosphorous sources and yeast extract in the medium on cell growth, synthesis and secretion of protease, and production of two major nonbiomass products, viz., acetic acid and ethanol. Cell growth and formation of the three nonbiomass products are hampered significantly under nitrogen, phosphorous, or oxygen limitation, with the cells being unable to grow in an oxygen-free environment. Improvement in protease production is achieved with respect to each culture parameter, leading in the process to 80% enhancement in protease activity over that attained using media reported in the literature. Results of a few fed-batch experiments with constant feed rate, conducted to examine possible enhancement in protease production and to further investigate repression of protease synthesis by excess of the principal carbon and nitrogen sources, are also discussed. The detailed investigation of stimulatory and repressory effects of simple and complex nutrients on protease production and metabolism of Bacillus firmus conducted in this study will provide useful guidelines for design of bioreactors for production of protease and bulk chemicals by this bacterium.
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    Biotechnology and Bioengineering 37 (1991), S. 512-518 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pseudomonas putida biofilms were developed on and biofilm accumulation rate data were obtained for the following two classes of support materials: charged surfaces and noncharged hydrophobic and hydrophilic surfaces. The effects of surface roughness and porosity on the rate of microbial attachment were also examined.Materials bearing a net positive or negative surface charge supported the greatest biofilm accumulation and the highest biofilm accumulation rate. Uncharged hydrophobic materials achieved the next greatest biofilm accumulation, averaging approximately 50% of the total biomass which was accumulated on the charged surface materials after 16 days. Uncharged hydrophilic materials supported very little biofilm development. In general, biofilm accumulation increased with decreased surface roughness. The effect of pore size on biofilm accumulation was not conclusive.The biofilm accumulation kinetics showed an exponential accumulation rate for the charged surfaces and an approximately linear accumulation rate for the hydrophobic materials. This difference in accumulation kinetics is consistent with proposed differences in the physicochemical mechanism governing attachment to these two types of surface materials.
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    Biotechnology and Bioengineering 35 (1990) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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    Biotechnology and Bioengineering 35 (1990), S. 976-982 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The concept of pore size distribution is incorporated into the Clark model of enzyme immobilization in the present study. This refined model predicted that in the case of small harmonic pore radius with the same surface area and porosity of the support, more enzyme could be loaded in a support with nonuniform pores than that with uniform pores. In comparing the enzyme loading efficiency of the support with two different pore size distributions, the one with Gaussian distribution had the greater amount of enzyme immobilized than the other one with Rajagopalan's distribution. Furthermore, more enzyme could be loaded in a support with wider Gaussian pore size distribution than that with narrower distribution. The immobilized enzyme profile in the solid support with pore size distribution displayed a stepwise pattern which differed appreciably from the sigmoidal profile predicted for the support with uniform pore size. This stepwise enzyme distribution profile became sigmoidal with decreasing hT or increasing k. The new model could be used for designing protocols for an enzyme immobilization process.
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    Biotechnology and Bioengineering 35 (1990), S. 1000-1005 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new method for the immobilization of microbial cells has been developed. Whole cells of Escherichia coli with aspartase activity were immobilized by capture on the surface of cross-linked poly(N-benzyl-4-vinylpyridinium bromide) containing styrene (BVPS resin), an insoluble pyridinium-type resin. When a suspension of the bacterial cells in buffer solution was passed through a glass column containing beads of BVPS resin, the cells were captured on the resin surface and formed an immobilized cell system. A fixed-bed column reactor containing 300 mg of the bacterial cells immobilized by capture on 10 g of BVPS resin beads was used for the preparation of L-aspartic acid from ammonium fumarate. Continuous operation of tne bioreactor produced L-aspartic acid in a quantitative yield when the influent substrate concentration was 0.1M and the flow rate was 0.41-0.83 bed volumes per hour at pH 7.4-7.7 at 30°C.
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    Biotechnology and Bioengineering 35 (1990), S. 1024-1033 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A mathematical model for an ideal chemostat in which one microbial population feeds on another and where Monod's model is used for the specific growth rates of both populations predicts a less stable behavior for the system than the one observed experimentally. Various factors have been proposed as being the reason for the increased stability of such systems. In this work, the effect of spatial heterogeneity on the dynamics of the microbial feeding interaction is studied. It is concluded that spatial heterogeneity has a stabilizing effect on the system. This effect combined with other factors could be the reason for the increased stability observed in systems where a microbial feeding interaction occurs.
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    Biotechnology and Bioengineering 35 (1990), S. 1078-1087 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An oxygen microsensor was used to measure internal oxygen profiles in biocatalyst particles of different diameter and activity. The particles were made of agarose gel and contained an oxygen reducing enzyme, L-lactate mono-oxygenase. The kinetics of the enzyme could be well described by the Michaelis-Menten equation. From the internal substrate concentration profile the intrinsic kinetic parameters were determined by means of fitting a simulated profile to the measurements, using Marquardt's algorithm. The intrinsic kinetic parameters found following this procedure appeared to be independent of particle radius or enzyme loading used, proving the method to be reliable. These parameters were also compared with the kinetic parameters of the free enzyme which were determined in a biological oxygen monitoring system. The intrinsic kinetic parameters showed a decrease with a factor 2.3 for Vm value and with a factor 2.7 for the Km value compared to the parameters for the free enzyme. From this the conclusion can be drawn that the immobilization as such or the carrier material not only can have an effect on the maximum intrinsic conversion rate (Vm) but also on the affinity of the enzyme (Km) for oxygen.
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    Biotechnology and Bioengineering 35 (1990), S. 1120-1124 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A gene for ethionine resistance isolated from the yeast Saccharomyces cerevisiae DKD-5D-H conferred on the yeast cells resistance to seleno-L-methionine and capability to produce S-adenosyl-L-methionine in the cells. An enzymatic study of the L-methionine synthetic pathway of L-methionine proto- and auxotrophs and in dried yeast cells with or without the gene suggested that the cloned gene for ethionine resistance is responsible for the activity of S-adenosyl-L-methionine synthase. To produce S-adenosyl-L-methionine by yeast cells transformed with the ethionine resistance gene, some culturing conditions were determined.
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    Biotechnology and Bioengineering 35 (1990), S. 1135-1144 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of fluid flow and laminar shear on bacterial uptake was examined under conditions representative of the fluid environment of unattached and attached cells in wastewater treatment bioreactors. Laminar shear rates below 50 s-1 did not increase leucine uptake by suspended cultures of Zoogloea ramigera. However, leucine uptake by cells fixed in a flow field of ∼ 1 mm s-1 was 55-65% greater than uptake by suspended cells. Enhanced microbial uptake with advective motion is consistent with mass transfer rates calculated using Sherwood number correlations. Advective flow increases microbial uptake by increasing collisions between substrate molecules and cells through compression of the concentration boundary layer surrounding a cell. The rate of leucine uptake suggests that binding proteins used to transport leucine into the cell can occupy approximately 1% of the cell surface area.
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    Biotechnology and Bioengineering 35 (1990), S. 1169-1173 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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    Biotechnology and Bioengineering 36 (1990), S. 1-11 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The constitutive cytoplasmic expression in E. coli of human growth hormone (hGH) with different N-terminal extensions (3 or 4 amino acids) has been studied. These hGH precursors were used for in vitro cleavage to obtain the mature, authentic hormone. Small changes in the amino acid extensions of the hGH precursors led to three-fold differences in specific expression rates. The specific expression rate of the hGH precursors was inversely proportional to the ratios of the specific growth rates of plasmid containing and plasmid free cells (μ+/μ-) and also to the genetic stability. To ensure a satisfactory genetic stability in production fermentors, an hGH precursor with a moderate expression efficiency was chosen.The medium composition and growth conditions were studied, resulting in the choice of a glucose fed batch fermentation process using a complex medium. In this process a yield of 2000 mg/L of met-ala-glu-hGH (MAE-hGH) was obtained. The fermentation process comprised a glucose-limited growth phase followed by a second phase with increased glucose feed and exhaustion of phosphate from the medium. The second phase is characterized by an MAE-hGH production, whereas further biomass formation is blocked. High concentrations of glucose led to reduced specific expression of MAE-hGH - the specific and total yield in batch glucose fermentations is only about 30% of the yield in optimized fed batch fermentations. The physiological background for this was investigated. Chemostat experiments showed that the glucose concentration and the metabolic condition of the cells - i.e. with or without formation of acetate - was not critical per se in order to obtain a high specific yield of MAE-hGH. Therefore it is unlikely that formation of MAE-hGH is catabolite repressed by glucose. Furthermore it was shown that the specific production rate of MAE-hGH was independent of the specific growth rate and it was further demonstrated that the decrease in expression efficiency in glucose batch fermentation was a result of an inhibitory effect of acetic acid. In batch fermentations this inhibitory effect was enhanced by a salt effect caused by increased consumption of acid and base used to control pH. The identity of the acid and the base used are not important in this context.From studies of the expression of other proteins in E. coli. with constitutive as well as inducible promoters we conclude that glucose fed batch processes are often superior to batch processes in the production of heterologous proteins E. coli.
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  • 77
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    Biotechnology and Bioengineering 36 (1990), S. 47-54 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Immobilization of Saccharomyces cerevisiae ATCC 834 within alginate beads enhances microbiological conversion of benzaldehyde to L-phenylacetyl carbinol (L-PAC), a precursor employed for synthesis of L-ephedrine. Yields of 90% L-PAC on benzaldehyde (initially 0.6% in medium) were obtained with immobilized cells, in contrast to about 10% with free cells which tend to form pellets in the presence of benzaldehyde. The predominant favorable action of immobilization appears to be a reduction in the toxic or inhibitory effects of benzaldehyde. With an initial benzaldehyde concentration of about 0.6% in the medium the optimum cell mass concentration was observed to be about 28 g cell mass (immobilized) per liter of medium.
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  • 78
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    Biotechnology and Bioengineering 36 (1990), S. 92-96 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fluctuations in pH and head-space pressure in a fermentor introduce temporary changes in off-gas CO2 concentrations. These changes are quantified using a simple model based on kinetics of CO2 hydration and gas-liquid mass transfer. The model is verified experimentally. An eigenvalue analysis of the model indicates that mass transfer is the parameter which controls the dynamics of CO2 equilibration.
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  • 79
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    Biotechnology and Bioengineering 36 (1990), S. 104-108 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 80
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    Biotechnology and Bioengineering 36 (1990), S. 124-134 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Recombinant cell growth and protein synthesis by a recombinant Escherichia coli under various inducing conditions are compared to the predictions of a mathematical model. The mathematical model used was a combination of two literature models: (1) an empirical kinetic model for recombinant growth and product formation and (2) a genetically structured model of the lac promoter-operator on a multicopy plasmid. The experimental system utilized was recombinant E. coli CSH22 bearing the temperature-sensitive plasmid pVH106/172, which codes for the synthesis of β-galactosidase and the other lac operon genes under the control of a lac promoter. Mathematical model predictions for recombinant β-galactosidase yield and specific growth rate were compared with fermentation measurements of these same quantities for conditions of chemical induction with cyclic AMP and IPTG, copy number amplification (by shifting culture temperature), and combined chemical induction and copy number amplification. The model successfully predicted experimental product yields for most cases of chemical induction even though the product yields varied from 0.34 × 103 to 1500 × 103 units/g cell mass. The kinetic model also correctly predicted a decline in the specific growth rate with increasing levels of plasmid and recombinant protein. The model was less successful at predicting product amplification at high copy numbers. A comparison of model predictions and experimental results was also used to investigate some of the assumptions used in constructing the mathematical models.
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  • 81
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    Biotechnology and Bioengineering 37 (1991), S. 927-935 
    ISSN: 0006-3592
    Keywords: Bacillus subtilis ; recombinant plasmid ; deletion rate ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Recombinant plasmid pCEDS is structurally unstable in Bacillus subtilis cultures. We have previously shown that stability can be independently increased by changing from a complex medium supporting high growth rates to a chemically-defined medium supporting a lower growth rate and removal of a 4.77-kb EcoRI fragment from pCED3 to give plasmid YS1. Further stabilization was achieved by combining the two approaches. In the present work, we show that the stabilization of the plasmid-encoded LacZ+ phenotype can be explained solely by the effect on the growth rate ratio between cells containing modified and parental plasmids. By using modified stability experiments (where a single cell rather than a suspended colony was used to initiate growth), independent growth rate measurements, and a simple mathematical model, we can describe the kinetics of the loss of the LacZ+ phenotype in terms of two variables, α and p (where α is the ratio of growth rates between modified and parental cells, and p is the probability of obtaining modified cells from parental cells). Under the conditions tested, the average values of α were 1.52 for cultures growing in complex medium, 1.28 for cultures growing in defined medium, and 1.18 for cultures containing the modified plasmid pYS1 growing in complex medium. The calculated p values ranged between 10-8 and 10-10 under all conditions. Plasmid (pYS137) was used to directly estimate plasmid deletion rates in B. subtilis and it showed a rate between 5 × 10-8 and 1.1 × 10-9 deletions/cell/generation. In contrast to B. subtilis, there were no detectable differences in growth rates between Escherichia coli strains harboring plasmid pCEDS and plasmid-free cells. These results explain the observed stability of pCEDS in E. coli cultures and indicate that readily detected instability in B. subtilis cultures can be the result of rare deletion events.
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  • 82
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    Biotechnology and Bioengineering 37 (1991), S. 960-966 
    ISSN: 0006-3592
    Keywords: bioreactors ; oxygen transfer ; fermentation ; mycelial fermentation ; mass transfer ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The oxygen transfer in bioreactors with slurries having a yield stress was investigated. The volumetric mass transfer coefficients in a 40-L bubble column with simulated fermentation broths, the Theological properties of which were represented by the Casson model, were measured. Experimental data were compared with a theoretical correlation developed on the basis of a combination of Higbie's penetration theory and Kolmogoroff's theory of isotropic turbulence. Comparisons between the proposed correlation and data for the simulated broths show good agreement. The mass transfer data for actual mycelial fermentation broths reported previously by the authors were re-examined. Their Theological data was correlated by the Bingham plastic model. The oxygen transfer rate data in the mycelial fermentation broths fit the predictions of the proposed theoretical correlation.
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  • 83
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    Biotechnology and Bioengineering 37 (1991), S. 989-993 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 84
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    Biotechnology and Bioengineering 37 (1991), S. 998-1003 
    ISSN: 0006-3592
    Keywords: Arthrobacter simplex ; controlled ultrasonic irradiation ; microbial conversion ; ultrasound facilitated diffusion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Dehydrogenation of hydrocortisone by agitated suspensions of free and immobilized cells of Arthrobacter simplex ATCC 6946 was investigated under controlled ultrasonic irradiation at a frequency of 20 kHz. The microbial conversion was optimized first with respect to mechanical agitation and subsequently with respect to an additionally superimposed sonication. The optimization of ultrasound intensity and its mode of application were established at a level which maintained the structural integrity of the cells as well as their biocatalytic activity. Various regimes of ultrasound at power densities of 0.030-0.120 W/mL were applied in systems of soluble (0.4 g/L) and excess (1 g/L) hydrocortisone and only a moderate enhancement of the bioconversion by free cells was observed. This result was explained by a better ultrasound-induced dispersal of microscopic clumps of cells and self-adhering clusters of the steroids. However, a quite significant enhancement effect was obtained in bioconversion systems of soluble substrate by gel-entrapped cells. This enhancement was explained by a phonophoretic effect associated with ultrasound-facilitated diffusion of the substrates - oxygen and hydrocortisone - within the gel beads.
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  • 85
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    Biotechnology and Bioengineering 37 (1991), S. 1037-1042 
    ISSN: 0006-3592
    Keywords: biodegradation ; organic solvents ; biooxidation ; bacteria ; competition interactions ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Much more information concerning the biodegradation kinetics of mixtures of common industrial chemicals, such as organic solvents, needs to be gathered before wastewater biotreatment process performance can become a matter of design. Here, the biooxidation of a solvent mixture comprizing methanol, acetone, isopropanol, and methylene chloride is examined. The fact that the enrichment culture obtained comprized only two solvent-utilizing strains, together with only minor percentages of nonsolvent utilizing satellite strains, was contrary to the theory of microbial competition. In addition, the complex relationship between the two solvent-utilizing strains indicates that further work is necessary on the pathways involved in isopropanol and acetone biooxidation and on the effects of operating conditions on the fluxes along such pathways.
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  • 86
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    Biotechnology and Bioengineering 37 (1991), S. 1066-1075 
    ISSN: 0006-3592
    Keywords: Resonance energy transfer ; papain ; concanavalin A ; molecular model ; monovalent ligand and receptor ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A stochastic model is described that predicts the degree of singlet/singlet energy transfer in complexes formed between monovalent ligands and monovalent receptors. The modeling approach is intended to serve as an analytical tool for approximating the level of fluorescence quenching that can be expected to occur in fluorescently labeled monovalent ligands and receptors that are bound together in complexes. This approach has utility in areas such as modeling protein/protein interactions and designing fluorescence energy transfer assays.Using the crystallographic data for papain (monovalent ligand ) and concanavalin A (monovalent receptor ) along with a molecular graphics computational package the ligand and receptor were docked together to form a ligand/receptor complex. The intermolecular distances between the lysine resides of the ligand and receptor were then estimated, receptor complex was calculated assuming a value for the characteristic length R0 of the donor/acceptor pair. Results from the stochastic model were used to calculate the level of fluorescence quenching one would expect for a resonance energy transfer competition assay based on the monovalent ligand/pair.Three key assumptions were made during the model development. First, all lysine resides for the ligand and receptor were equally reactive with the dye molecules so the stoichiometry of the donor and acceptor chromophores was governed by a binomial distribution. Second, the dye molecules were located at the α-carbon position for each reactive lysine residue. Finally, in the energy transfer competition assay, it was assumed that equilibrium existed between the ligand, receptor, and competing hapten at all times. Based on these assumptions, results are presented that indicate the maximum energy transfer for the monovalent papain/concanavalin. A complex is strongly dependent on the number of acceptor chromophores and on the value of R0. Results are also presented on the approximate level of fluorescence quenching that may occur in a competition assay based on the papin/pConA complex. Lastly, a strategy is discussed for maximizing the dynamic range and linearity of energy transfer assays by optimizing several key design variables.
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  • 87
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    Biotechnology and Bioengineering 37 (1991), S. 1108-1116 
    ISSN: 0006-3592
    Keywords: organic acids ; Aspergillus flavus ; molar yield ; stirred fermentor ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Effects of various nutritional and environmental factors on the accumulation of organic acids (mainly L-malic acid) by the filamentous fungus Aspergillus flavus were studied in a 16-L stirred fermentor. Improvement of the molar yield (moles acid produced per moles glucose consumed) of L-malic acid was obtained mainly by increasing the agitation rate (to 350 rpm) and the Fez+ ion concentration (to 12 mg/L) and by lowering the nitrogen (to 271 mg/L) and phosphate concentrations (to 1.5 mM) in the medium. These changes resulted in molar yields for L-malic acid and total C4 acids (L-malic, succinic, and fumaric acids) of 128 and 155%, respectively. The high molar yields obtained (above 100%) are additional evidence for the operation of part of the reductive branch of the tricarboxylic acid cycle in L-malic acid accumulation by A. flavus. The fermentation conditions developed using the above mentioned factors and 9% CaCO3 in the medium resulted in a high concentration (113 g/L L-malic acid from 120 g/L glucose utilized) and a high overall productivity (0.59 g/L h) of L-malic acid. These changes in acid accumulation coincide with increases in the activities of NAD+-malate dehydrogenase, fumarase, and citrate synthase.
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    Biotechnology and Bioengineering 38 (1991), S. 30-36 
    ISSN: 0006-3592
    Keywords: lipase-catalyzed hydrolysis ; 2-naphtyl ester ; biphasic system ; interfacial reaction ; two-film model ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The authors measured the rate of hydrolysis of the homologs of 2-naphtyl ester by using a Lewis cell with constant interfacial area to elucidate the kinetic mechanism of the lipase-catalyzed hydrolysis in biphasic system. On the basis of the two-film model, it was found from the analysis of experimental results that the hydrolysis of these substrates proceeds at the interface between the aqueous and organic phases. The interfacial reaction rate could be correlated by Michaelis-Menten mechanism. The values of the rate constant and the Michaelis constant were almost independent of the kinds of 2-naphtyl ester. The values of the interfacial kinetic parameters for 2-naphtyl ester were much greater than those for the hydrolysis in the aqueous phase.
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  • 89
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    Biotechnology and Bioengineering 38 (1991), S. 65-74 
    ISSN: 0006-3592
    Keywords: Succinoglucan ; Agrobacterium radiobacter ; Structured model ; ATP ; cofactors balances ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Wild-type Agrobacterium radiobacter NCIB 9042 has been cultivated in batch cultures on a synthetic medium which was adapted for growth and succinoglucan production. Experiments were carried out in a 4-L stirred-tank aerated reactor. Glucose, biomass, polysaccharide, protein, and inorganic- and organic-nitrogen concentrations were measured, and oxygen consumption and CO2 production rates were obtained by a gas-balance technique. Nitrogen balance shows that inorganic nitrogen is entirely recovered into proteins. The carbon balance is satisfied with in ±5%. Stoichiometric equations for biomass growth and succinoglucan synthesis were established. The biosyntheticpolymer pathways including ATP and cofactor consumption were investigated. From previous studies, a (P/O) value of 1.66 is selected for oxygen sufficient cultures. The actual ATP requirements of 25.4 mmol ATP/g succinoglucan (38.5 mol ATP/mol succinoglucan), determined by a metabolic analysis, is 2.39 times the stoichiometric value. Experimental results were modeled by a system of differential equations. The exponential growth phase was described by a nitrogen-limited Monod equation. Subsequent succinoglucan synthesis followed a slightly modified Luedeking-Piret relation partitioning internal and external polysaccharide. Experimentally determined coefficients are compared with published results for continuous culture of A. radiobacter NCIB 11883.
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  • 90
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Sensors for the simultaneous determinations of sucrose and glucose, lactose and glucose, and starch and glucose were prepared by a combination of the enzyme system shown below and an oxygen electrode: The mechanism for separating the substrates with the proposed sensors is based on the time lag arising from reaction and diffusion. Invertase, β-galactosidase, amyloglucosidase, mutarotase, and glucose oxidase were covalently immobilized on triacetyl cellulose membranes containing 1,8-diamino-4-aminomethyloctane. A glucose oxidase membrane, mutarotase membrane, three sheets of triacetyl cellulose membranes, and invertase, or β-galactosidase or amyloglucosidase membrane were placed in that order on the tip of the oxygen electrode. Calibration curves for sucrose, lactose, and starch were linear up to 40 mM, 60-180 mM, and 10%, respectively. The simultaneous determination of sucrose and glucose, lactose and glucose, and starch and glucose was possible when the amount of glucose coexised was in the range of 2-16% sucrose, 2.8-8.3% lactose, or 0.1-1% starch. The relative errors were ±4% for sucrose and ±3% for lactose in 100 assays. The starch sensor was reused only five times. Each enzyme membrane was fairly stable for more than 10 days.
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  • 91
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    Biotechnology and Bioengineering 38 (1991), S. 127-134 
    ISSN: 0006-3592
    Keywords: Bacillus subtillis ; binding free energy ; Adsorption isotherm ; monolayer adsorption process ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The goal of this work was to characterize the adsorption of Bacillus subtills α-amylase onto crystalline starchy materials of the B-type polymorph. Monodisperse spherulitic particles (R ż6; 5.0 μm), essentially resistant to α-amylolysis at 25°C were prepared from short amylose chains (DPn ≈ 15). The α-amylase adsorbed specifically onto the spherulites, and adsorption was found to be a prerequisite step for hydrolysis. Adsorption was inhibited by the presence of maltose and maltotriose in the reaction mixture. Adsorption isotherm of the enzyme on the particles showed a well developed plateau of 1.62 μg/cm2 at 25°C corresponding to a monolayer adsorption process. The binding free energy calculated from the initial slope of the isotherm was ΔG ≈ -20.7 kJ/mol. This is smaller than published values for the binding of α-amylase to soluble amylosic chains (ΔG 〈 -30 kJ/mol).
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  • 92
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    Biotechnology and Bioengineering 36 (1990), S. 617-622 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Lipase from Pseudomonas fluorescens biotype I was immobilized by adsorption of anion exchange resin using glutaraldehyde to enhance the adsorption. The activity yield of the immobilized lipase was very low (below 1%) when lipase activity was measured using emulsion substrate. The activity yield was 10-70% when lipase activity was measured using non-emulsion substrate. Countercurrent reactors for hydrolysis of oil using non-emulsion substrate were studied. A fluidized bed reactor was found to be superior to a fixed bed one since in a fixed bed reactor the separation rate of the two layers was slow and the flow rate of the reactor had to be slower than the separation rate. A fluidized bed reactor system equipped with settling compartments and stirring compartments was devised. Continuous lipolysis at 60 °C and continuous separation of oily product and water soluble product were performed. After continuous operation for more than 3 months, 70% of the initial activity of the immobilized lipase was observed at the end of the reaction.
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  • 93
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    Biotechnology and Bioengineering 37 (1991) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 94
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    Biotechnology and Bioengineering 37 (1991), S. 1004-1009 
    ISSN: 0006-3592
    Keywords: esterification ; immobilized enzymes ; enzymes in organic solvents ; enzyme activity and stability ; continuous enzymatic synthesis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Various esters were synthesized in nearly anhydrous hexane from alcohols and carboxylic acids using a lipase from Candida cylindracea. The enzyme was immobilized on a nylon support and protein loadings as high as 10 mg/g were obtained. The activity of the immobilized enzyme was maximum in a range of temperatures from 25 to 37°C. Ethylpropionate was formed from ethanol and propionic acid at a rate of 0.017 mol/h g immobilized protein. Different esters were formed at comparable rates and equilibrium conversions could generally be approached in less than 10 h in a batch reaction system. The immobilized lipase catalyst was quite stable and retained about one third of the initial activity after repeated experiments during the course of 72 days. A stirred tank continuous flow reactor was used successfully for the continuous production of esters.
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  • 95
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    Biotechnology and Bioengineering 37 (1991), S. 1029-1036 
    ISSN: 0006-3592
    Keywords: immobilization ; protein production ; continuous culture Saccharomyces cerevisiae ; plasmid stability ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The Saccharomyces cerevisiae strain Mc16/p520 has an unstable plasmid, p520, which directs production of a wheat α-amylase. The effects of immobilizing this microorganism on the plasmid stability and the specific productivity of the secreted α-amylase were investigated. Small gelatin beads were used as the support in both fluidized and packed bed configurations, and the yeast cells were attached by covalent cross-linking with glutaraldehyde. These data were then compared to those for nonimmobilized, suspension cells.Plasmid stability was increased for the immobilized cells during continuous culture at dilution rates both above and below washout. Continuous suspension cultures were not stable and rapidly lost the plasmid. Immobilization caused an increase in specific and volumetric productivity during continuous culture, with a packed bed design resulting in the highest specific productivity.
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  • 96
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    Biotechnology and Bioengineering 37 (1991), S. 1050-1053 
    ISSN: 0006-3592
    Keywords: hybridomas ; immobilization ; oxygen ; respiration ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Oxygen consumption by hybridoma cells immobilized in 1- and 3.9-mm-diameter calcium alginate beads was measured. The entrapped cells consumed oxygen at about 10 μmol/min per 109 cells, regardless of the bead size and cell loading. In contrast, the same cells in suspension culture respire at specific rates of 3-8 μmol/min per 109 cells (depending on the cell density). The growth rate of the immobilized cells was significantly reduced, while specific antibody production was comparable to that of free cells.
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  • 97
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In order to produce a product with a high content of maltotetraose, dual-enzyme systems composed of immobilized maltotetraose-forming amylase (G4-forming amylase) and pullulanase were studied. The thermostability of individually immobilized enzymes was examined in continuous operation; studies revealed that the enzyme immobilized on “Chitopearl” was much more stable than that immobilized on Diaion HP-50. The effects of operating conditions on the stability of G4 forming amylase immobilized on “Chitopearl” were examined to confirm that the apparent half-life data could be arranged using the immobilized enzyme stability factor, fs. As for the dual immobilized enzyme system, six methods of usage were considered, with five yielding a 7-10% (w/w) higher content of maltotetraose product than the single-enzyme system. The effects of operating conditions on the maltotetraose production reaction were examined to confirm that the maltotetraose content of the products could be analyzed using the specific space velocity,SSV. In dual immobilized enzyme systems, pullulanase immobilized on the same carrier as the G4-forming amylase was found to be more stable than pullulanase immobilized on separate carriers. The effectiveness of using immobilized pullulanase along with the G4-forming amylase was confirmed from constant-conversion operations in which the maltotetraose content in the product was kept at 50% (w/w) in laboratory-scale experimentation.
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  • 98
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    Biotechnology and Bioengineering 36 (1990), S. 821-825 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In Part II, the process utility of a thermotolerant methylotrophic bacterium is evaluated with respect to its dynamic response when various substrate or nutrient pulses are imposed on it while growing under steady-state conditions in a chemostat. The pulses investigated were methanol pulses on methanol-, methanol/formaldehyde-, and dual methanol/ammonia-limited cultures and an ammonia pulse on a severely nitrogen (ammonia)-limited culture. The results obtained, although exemplifying the complex biochemistry of such bacteria, clearly demonstrate the bacteriums flexibility and versatility in handling process transients. Its lack of fastidiousness in unsteady state continuous culture make it a most promising candidate for inclusion in process cultures for elevated temperature industrial wastewater treatment.
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    Biotechnology and Bioengineering 36 (1990) 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 36 (1990), S. 879-886 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A hollow-fiber membrane reactor was designed and constructed to allow perfusion of entrapped, dense Escherichia coli cells with nutrient medium during examination of cell metabolism using nuclear magnetic resonance (NMR) spectroscopy. Phosphorus-31 NMR spectra of the perfused cells included peaks for nucleoside di- and triphosphates, sugar phosphates, and pH-sensitive peaks for inorganic phosphate. The observed intensity of the lumenal inorganic phosphate peak was found to depend on flow rate, ruling out the use of this peak as a concentration reference. Absolute intracellular pH values obtained from NMR measurements were found to be accurate to 0.2 pH units due to uncertainties in intracellular ionic concentrations. Relative pH values, however, were found to be sensitive to cell energetic status. The response of E. coli intracellular pH following a shift to carbon starvation medium was monitored with a resolution of 3 min. Use of a hollow-fiber reactor for cell containment and perfusion during NMR spectroscopy enables metabolic experiments of longer duration and of greater variety than is possible using standard, nonperfused sample tubes.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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