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  • Articles  (128)
  • Escherichia coli  (67)
  • fermentation  (66)
  • 2010-2014
  • 1990-1994  (118)
  • 1985-1989  (10)
  • 1950-1954
  • Process Engineering, Biotechnology, Nutrition Technology  (128)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 15 (1994), S. 169-176 
    ISSN: 1573-0778
    Keywords: Flaviviridae ; BDV ; fermentation ; metabolism ; vaccines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Vaccines on the basis of mammalian cell cultures are of major importance for human and animal health. Therefore efforts are undertaken for the improved production of more effective vaccines. Of course, the main purpose of all these approaches is to save lives and improve the quality of life for human beings. However, there is also some remarkable effort in the food industry and the associated animal production, especially in the case of some Flaviviridal viruses (BVD), where〉80% of all cattle herds are found to be infected. These viruses can cause tremendous economic losses of calfs and embryos (Ames, 1990). Because of these facts, there is a continuous endeavour for improving the manufacturing of therapeutics or preventing agents such as vaccines for the treatment of cattle. The competitive economic situation and the specific market demands still require effective and high yield production methods, especially in the case of one of the most widespread viral diseases in cattle like BVD (Ames, 1990). We have succeeded in establishing an improved method for the production of BVD on the basis of a continuous fermentation mode, that consist of modifications of the corresponding process and media improvements.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 15 (1994), S. 271-279 
    ISSN: 1573-0778
    Keywords: Bioreactor ; control ; computer ; fermentation ; perfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In the last 10 years, new assignments and the special demands of mammalian cells to the culture conditions caused the develoepment of complex small scale fermentation setups. The use of continuous fermentation and cell retention devices requires appropriate process control systems. An arrangement for control and data-acquisition of complex laboratory-scale bioreactors is presented. The fundamental idea was the usage of a standard personal computer, which is connected to pumps, valves and sensors via ADA-transformation. The possibility of free programming allowed the development of user-oriented software, especially designed for the far-reaching requirements of a university laboratory in the field of animal cell culture. Control of aeration, pumps, data-acquisition and data-storage are combined within one program, which allows the automation of standard operations like measurement of kLa- or OTR-values. Pump control algorithms for all common fermentation strategies (batch, fed batch, chemostat, perfusion) are included and can be selected any time during cultivation. Oxygen partial pressure and pH are controlled via direct digital control (ddc), providing simple adaption of control parameters and set points to current fermentation conditions.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    World journal of microbiology and biotechnology 10 (1994), S. 410-413 
    ISSN: 1573-0972
    Keywords: Enterobacteriaceae ; fermentation ; lactic acid bacteria ; maize ; mawè ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Lactic acid bacteria increased from 3.2 × 106 and 1.6 × 107 c.f.u./g (wet wt) to 2 × 109 and 1.6 × 109 c.f.u./g after 12 to 24 h of fermentation of home-produced mawè (a dough produced from dehulled maize) and commercial mawè, respectively. In commercial mawè, the yeast count increased from 1.3 × 105 to 2.5 × 107 c.f.u./g after 48 h of fermentation before decreasing, whereas in the home-produced mawè it increased from 2.5 × 104 to 3.2 × 107 c.f.u./g after 72 h of fermentation; the dominant yeasts were mainly Candida krusei, although C. kefyr, C. glabrata and Saccharomyces cerevisiae were also present. Enterobacteriaceae counts increased slightly during the initial stage ofthe fermentation, but decreased below the detection level after 24 to 48 h. Enterobacter cloacae was mostly found in commercial mawè and Escherichia coli mostly in homeproduced mawè.
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  • 4
    Electronic Resource
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    Springer
    World journal of microbiology and biotechnology 10 (1994), S. 346-347 
    ISSN: 1573-0972
    Keywords: Chromatography ; Escherichia coli ; plasmid ; transfection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A simple procedure to obtain plasmid preparations, suitable for transfecting mammalian cell lines using a calcium phosphate co-precipitation technique, is described. The protocol is based on the purification of plasmid DNA by double gel-filtration chromatography on Sephacryl S-1000 and additional slight modifications to the original transfection procedure. The purity of plasmid preparation was verified by analytical methods. The resulting preparation efficiently transfected NIH-3T3 cells.
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  • 5
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 43 (1994), S. 399-410 
    ISSN: 0006-3592
    Keywords: lac-based promoters ; Escherichia coli ; genetic control ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A model that describes induction of protein synthesis from lac-based promoters has been developed and incorporated into the single-cell model of Escherichia coli with transcriptional and translational modifications. Unlike previous models of lac-based promoters, this model allows a priori prediction of the intracellular parameters controlling transcription from lac-based promoters with only the extracellular levels of substrate and inducer as inputs. Because of the structural detail of the model, it is possible to simulate different genetic constructions for comparison, such as Laclq strains versus wild-type cells, or including lacl on a multicopy plasmid. Expression from lac to tac promoters is predicted to yield 5% and 30% of the total cellular protein, respectively, with a pBR322-type plasmid. The model predicts the experimental observation that the Laclq strain is not as fully induced as the wild-type strains, even at higher inducer concentrations. Additionally, the model predicts the right order of magnitude of protein production from lac and tac promoters when mechanisms for attenuation of transcription at lower translational efficiency are considered. Finally, the model predicts that for high copy number systems ribosomes become limiting in the synthesis of plasmid-encoded proteins. © 1994 John Wiley & Sons, Inc.
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  • 6
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 43 (1994), S. 978-986 
    ISSN: 0006-3592
    Keywords: butanol ; fermentation ; Clostridium acetobutylicum ; acetone ; ethanol ; pervaporation ; fed batch ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Extractive acetone-butanol-ethanol (ABE) fermentation was carried out successfully using pervaporation and a low-acid-producing Clostridium acetobutylicum B18. A pervaporation module with 0.17 m2 of surface area was made of silicone membrane of 240 μm thickness. Pervaporation experiments using make-up solutions showed that butanol and acetone fluxes increased linearly with their concentrations in the aqueous phase. Fickian diffusion coefficients were constants for fixed air flow rates, and increased at higher sweep air flow rates. During batch and fed-batch fermentations, pervaporation at an air flow rate of 8 L/min removed butanol and acetone efficiently. Butanol concentration was maintained below 4.5 g/L even though Clostridium acetobutylicum B18 produced butanol steadily. Pervaporation could not remove organic acids efficiently, but organic acids did not accumulate because strain B18 produced little organic acid and recycled added organic acids efficiently. With pervaporation, glucose consumption rate increased compared to without pervaporation, and up to 160 g/L of glucose was consumed during 80 h. Cell growth was not inhibited by possible salt accumulation or oxygen diffusion through the silicone tubing. The culture volume was maintained relatively constant during fed-batch operation because of an offsetting effect of water and product removal by pervaporation and addition of nutrient supplements. © 1994 John Wiley & Sons, Inc.
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  • 7
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    Biotechnology and Bioengineering 44 (1994), S. 132-139 
    ISSN: 0006-3592
    Keywords: glycogen ; Escherichia coli ; cell growth ; acetate ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Excessive production of acetate is a problem frequently encountered in aerobic high-cell-density fermentations of Escherichia coli. Here, we have examined genetic alterations resulting in glycogen overproduction as a possible means to direct the flux of carbon away from the acetate pool. Glycogen overaccumulation was achieved either by using a regulatory glgQ mutation or by transforming cells with a plasmid containing the glycogen biosynthesis genes glgC (encoding ADPG pyrophosphorylase) and glgA (encoding glycogen synthase) under their native promoter. Both strategies resulted in an approximately five-fold increase in glycogen levels but had no significant effect on acetate excretion. The glgC and glgA genes were then placed under the control of the isopropyl---D-thiogalactopyranoside (IPTG) inducible tac promoter, and this construct was used to stimulate glycogen production in a mutant defective in acetate biosynthesis due to deletion of the ack (acetate kinase) and pta (phosphotransacetylase) genes. If glycogen overproduction in the ack pta strain was induced during the late log phase, biomass production increased by 15 to 20% relative to uninduced controls. Glycogen overaccumulation had a significant influence on carbon partitioning: The output of carbon dioxide peaked earlier than in the control strain, and the levels of an unusual fermentation byproduct, pyruvate, were reduced. Exogenous pyruvate was metabolized more rapidly, suggesting higher activity of gluconeogenesis or the tricarboxylic acid (TCA) cycle as a result of glycogen overproduction. Potential mechanisms of the observed metabolic alterations are discussed. Our results suggest that ack pta mutants over producing glycogen may be a suitable starting point for constructing E. coli strains with improved characteristics in high-cell-density fermentations. © 1994 John Wiley & Sons, Inc.
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  • 8
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 43 (1994), S. 275-285 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; amino acids ; linear optimization ; metabolic fluxes ; metabolic engineering ; culture stability ; oxygen ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The simultaneous growth and product formation in a microbial culture is an important feature of several laboratory, industrial, and environmental bioprocesses. Metabolic burden associated with product formation in these bioprocesses may lead to growth advantage of a nonproducing mutant leading to a loss of the producing population over time. A simple population dynamics model demonstrates the extreme sensitivity of population stability to the engineered productivity of a strain. Here we use flux balance analysis to estimate the effects of the metabolic burden associated with product secretion on optimal growth rates. Comparing the optimal growth rates of the producing and nonproducing strains under a given processing condition allows us to predict the population stability. In order to increase stability of an engineered strain, we determine processing conditions that simultaneously maximize the growth rate of the producing population while minimizing the growth rate of a nonproducing population. Using valine, tryptophan, and lysine production as specific examples, we demonstrate that although an appropriate choice of oxygenation may increase culture longevity more than twofold, total production as governed by economic criterion can be increased by several orders of magnitude. Choice of optimal nutrient and oxygen supply rates to enhance stability is important both for strain screening as well as for culture of engineered strains. Appropriate design of the culture environment can thus be used to enhance the productivity of bioprocesses that use engineered production strains. © 1994 John Wiley & Sons, Inc.
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  • 9
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    Biotechnology and Bioengineering 43 (1994), S. 314-320 
    ISSN: 0006-3592
    Keywords: Spongiococcum exetricicum ; fed-batch fermentation ; fermentation ; microalgae fermentation ; feedback control ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Optimization of cellular productivity of an industrial microalgae fermentation was investigated. The fermentation was carried out at Coors Biotech Products Company, Fort Collins, Colorado. A mathematical model was developed based on the data collected from pilot plant test runs at different operating conditions. Pontryagin's maximum principle was used for determining the optimal feed policy. A feedback control algorithm was also studied for maximizing the cellular productivity. During continuous operation, the optimum dilution rate was determined by an adaptive optimization scheme based on the steepest descent technique and a recursive least squares estimation of model parameters. A direct search algorithm was also applied to determine the optimum feed rate. Comparison of the theoretical results of the different optimization schemes revealed that the direct search algorithm was preferable because of its simplicity. The experimental results of real time application of the feedback algorithm agreed fairly well with those of the theoretical analyses. © 1994 John Wiley & Sons, Inc.
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  • 10
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    Biotechnology and Bioengineering 43 (1994), S. 388-398 
    ISSN: 0006-3592
    Keywords: ribosome synthesis ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Details of the mechanism for ribosome synthesis have been incorporated in the single-cell Escherichia coli model, which enable us to predict the amount of protein synthesizing machinery under different environmental conditions. The predictions agree quite well with available experimental data. The model predicts that ribosomal protein limitations are important when the translational apparatus is in high demand. Ribosomal RNA synthesis is induced by an increase in translational activity, which, in turn, stimulates ribosomal protein synthesis. However, as the demand increases still more, the ribosomal protein mRNA must compete with the plasmid mRNA for ribosomes, and the efficiency of translation of ribosomal proteins is reduced. © 1994 John Wiley & Sons, Inc.
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  • 11
    ISSN: 0006-3592
    Keywords: near-infrared spectroscopy ; fermentation ; on-line monitoring ; lactic acid ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A near-infrared (NIR) spectroscopy technique for the control of lactic acid fermentation process has been proposed. Lactic acid, glucose, and biomass concentrations were determined by the NIR spectroscopy method. The three parameters examined were closely correlated to the results obtained with classical laboratory procedures. Moreover, the conditions for the on-line utilization of the NIR spectroscopy measurement system were pointed out. The great versatility of the NIR spectroscopy should permit its use for other fermentation processes. © 1994 John Wiley & Sons, Inc.
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  • 12
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    Biotechnology and Bioengineering 44 (1994), S. 29-37 
    ISSN: 0006-3592
    Keywords: proteins, contaminant ; Escherichia coli ; Saccharomyces cerevisiae ; mammalian cell culture ; PAGE ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The protein components of three industrial recombinant expression systems: Escherichia coli, Saccharomyces cerevisiae, and a mammalian cell culture supernatant of CHO cells were characterized in terms of their molecular weight, isoelectric point, and relative surface hydrophobicity. Identification of individual proteins was done by reference to their position in protein band profiles by polyacrylamide gel electrophoresis (PAGE) of the crude material. This permitted a rapid and facile assignment of quantitative values for these three parameters to all the major protein components in these materials. Because it is the indigenous proteins in expression systems that will form the bulk of any impurities in the product, once the values of these parameters are known for any target recombinant protein, the data obtained will enable appropriate expression systems to be chosen for minimizing amounts of potential contaminants and reducing downstream processing requirements and costs. The data will also indicate which fractionation steps (i.e., charge, size or hydrophobicity-based) are likely to be best for distinguishing between target and contaminant proteins, thus aiding and early removal of the maximum quantities of undesired protein to bring subsequent bioseparation steps down in scale and cost and up in terms of efficiency. © 1994 John Wiley & Sons, Inc.
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  • 13
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    Biotechnology and Bioengineering 44 (1994), S. 539-548 
    ISSN: 0006-3592
    Keywords: cross-flow filtration ; Escherichia coli ; cell harvesting ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Cross-flow filtration of Escherichia coli strains was examined at the laboratory and pilot scales using Romicon 500,000 molecular-weight-cutoff hollow fiber membranes. Both the series resistance and macrosolute polarization models were employed to compare performances. Total dissolved solids content above 90 g/L and viscosity above 1.1 × 10-3 paċ s of cell-free culture media were found to decrease average filtration fluxes by over 60% both in the absence and presence of cells. Broth filtration with culture media of dissolved solids levels below 80 g/L were influenced to a greater extent by harvest cell density. The collodial nature of the complex nutrient responsible for the total solids increase affected prediction of filtration performance. Differences in strain filterability were observed with JM109 preferred over DH5 in high solids-containing media and RR1 preferred over JM109 in low dissolved solids-containing media. Their research demonstrates the importance of cell strain and media selection in the performance of early downstream processing steps. © 1994 John Wiley & Sons, Inc.
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  • 14
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    Biotechnology and Bioengineering 44 (1994), S. 1295-1305 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; fusion proteins ; Cellulomonas fimi ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fusion of the leader peptide and the cellulose-binding domain (CBD) of endoglucanase A (CenA) from Cellulomonas fimi, with of without linker sequences, to the N-terminus of alkaline phosphatase (PhoA) from Escherichia coli leads to the accumulation of significant amounts of the CBD-PhoA fusion proteins in the supernatants of E. coli cultures. The fusion proteins can be purified from the supernatants by affinity chromatography on cellulose. The fusion protein can be desorbed from the cellulose with water or guanidine-HCl. If the sequence IEGR in present between the CBD and PhoA, the CBD can be cleaved from the PhoA with factor Xa. The efficiency of hydrolysis by factor Xa is strongly in fluenced by the amino acids on either side of the IEGR sequence. The CBD released by factor Xa is removed by adsorption to cellulose. A nonspecific proteases from C. fimi, which hydrolyzes native CenA between the CBD and the catalytic domain, may be useful for removing the CBD from some fusion proteins. © 1994 John Wiley & Sons, Inc.
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  • 15
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    Biotechnology and Bioengineering 44 (1994), S. 1337-1347 
    ISSN: 0006-3592
    Keywords: poly-(3-hydroxybutyric acid) ; PHB ; Escherichia coli ; morphology ; plasmid ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A stable high-copy-number plasmid pSYL105 containing the Alcaligenes eutrophus polyhydroxyalkanoic acid (PHA) biosynthesis genes was constructed. This plasmid was transferred to seven Escherichia coli strains (K12, B, W, XL1-Blue, JM109, DH5α, and HB101), which were subsequently compared for their ability to synthesize and accumulate ploy- (3-hydroxybutyric acid) (PHB). Growth of recombinant cells and PHB synthesis were investigated in detail in Luria-Bertani (LB) medium containing 20 g/L glucose. Cell growth, the rate of PHB synthesis, the extent of PHB accumulation, the amount of glucose utilized, and the amount of acetate formed varied from one strain to another. XL1-Blue (pSYL105) and B (pSYL105) synthesized PHB at the fastest rate, which was ca. 0.2 g PHB/g true cell mass-h, and produced PHB up to 6-7 g/L. The yields of cell mass, true cell mass, and PHB varied considerably among the strains. The PHB yield of XL1-Blue (pSYL105) in LB plus 20 g/L glucose was as high as 0.369 g PHB/g glucose. Strains W (pSYL105) and K12 (pSYL105) accumulated the least amount of PHB with the lowest PHB yield at the lowest synthesis rate. JM109 (pSYL105) accumulated PHB to the highest extent (85.6%) with relatively low true cell mass (0.77 g/L). Considerable filamentation of cells accumulating PHB was observed for all strains except for K12 and W, which seemed to be due either to the overexpression of the foreign PHA biosynthesis enzymes or to the accumulation of PHB. © 1994 John Wiley & Sons, Inc.
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  • 16
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    Biotechnology and Bioengineering 44 (1994), S. 1228-1234 
    ISSN: 0006-3592
    Keywords: broth recycle ; water reuse ; Apiotrichum curvatum ; fermentation ; microbial lipid ; inhibition ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fermentation is a water-intensive process requiring treatment of large amounts of effluent broth. It is desirable to increase the ratio of product produced to the volume of effluent by minimizing the discharge of effluent from the fermentation process. A study of recycling spent fermentation process. A study of recycling spent fermentation broth for the subsequent fermentation was carried out with Apiotrichum curvatum an oleaginous yeast, as the working culture. Spent broth from a defined medium was recycled t replace as much as 75% of the water and salts for subsequent batches and this was repeated for seven sequential batches without affecting cell mass and lipid production. A 64% vlume reduction of wastewater was achieved in this manner. However, when using whey permeate as the medium, lipid production dropped after three consecutive recycle operations at 50% recycle, and after two consecutive recycle operations at 75% and 100% recycle. Accumulation of ions in the broth appeared to be responsible for the inhibition. An ion exchange step was able to eliminate the ion buildup and restore fermentation performance. © 1994 John Wiley & Sons, Inc.
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  • 17
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    Biotechnology and Bioengineering 44 (1994), S. 1271-1278 
    ISSN: 0006-3592
    Keywords: fermentation ; state estimation ; kalman filter ; multirate systems ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article discusses issues related to estimation and monitoring of fermentation processes that exhibit endogenous metabolism and time-varying maintenance activity. Such culture-related activities hamper the use of traditional, software sensor-based algorithms, such as the extended kalman filter (EKF). In the approach presented here, the individual effects of the endogenous decay and the true maintenance processes have been lumped to represent a modified maintenance coefficient, mc. Model equations that relate measurable process outputs, such as the carbon dioxide evolution rate (CER) and biomass, to the observable process parameters (such as net specific growth rate and the modified maintenance coefficient) are proposed. These model equations are used in an estimator that can formally accommodate delayed, infrequent measurements of the culture states (such as the biomass) as well as frequent, culture-related secondary measurements (such as the CER). The resulting multirate software sensor-based estimation strategy is used to monitor biomass profiles as well as profiles of critical fermentation parameters, such as the specific growth for a fed-batch fermentation of Streptomyces clavuligerus. © 1994 John Wiley & Sons, Inc.
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  • 18
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    Biotechnology and Bioengineering 43 (1994), S. 429-433 
    ISSN: 0006-3592
    Keywords: disinfection ; Escherichia coli ; water disinfection ; activated carbon fiber ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel electrochemical reactor employing activated carbon fiber (ACF) electrodes was constructed for disinfecting bacteria in drinking water. Escherichia coli adsorbed preferentially onto ACF rather than to carbon-cloth or granular-activated carbon. E. coli cells, which adsorbed onto the ACF, were killed electrochemically when a potential of 0.8 V vs. a saturated calomel electrode (SCE) was applied. Drinking water was passed through the reactor in stop-flow mode: 2mL/min for 12 h, o L/min for 24 h, and 1 mL/min for 6 h. At an applied potential of 0.8 V vs, SCE, viable cell concentration reamined below 30 cells/mL. In the absence of an applied potential, bacteria grew to a maximum concentration of 9.5 × 103 cells/mL. After continuous operation at 0.8 V vs. SCE, cells adsorbed onto the ACF could not be observed by scanning electron microscopy. In addition, chlorine in drinking water was completely removed by the reactor. Therefore, clean and efficient inactivation of bacteria in drinking water was successfully performed. © 1994 John Wiley & Sons, Inc.
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  • 19
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    Biotechnology and Bioengineering 43 (1994), S. 483-489 
    ISSN: 0006-3592
    Keywords: microbial fermentation control ; neural network simulation ; backpropagation ; network topology design ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article discusses the development of a prototype neural network-based supervisory control system for Bacillus thuringiensis fermentations. The input pattern to the neural network included the type of inoculum, operation temperature, pH value, accumulated process time, optical density in fermentation medium, and change in optical density. The output from the neural network was the predicted optical density for the next sampling time. The control system has been implemented in both a computer simulation and a laboratory fermentation experiment with promising results. © 1994 John Wiley & Sons, Inc.
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  • 20
    ISSN: 0006-3592
    Keywords: lignocellulose ; ethanol ; Klebisella oxytoca ; fermentation ; cellulase ; cellulose ; cellobiose ; biomass ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pretreatment of sugar cane bagasse is essential for a simultaneous saccharification and fermentation (SSF) process which uses recombinant Klebsiella oxytoca strain P2 and Genencor Spezyme CE. Strain P2 has been genetically engineered to express Zymomonas mobilis genes encoding the ethanol pathway and retains the native ability to transport and metabolize cellobiose (minimizing the need for extracellular cellobiase). In SSF studies with this organism, both the rate of ethanol production and ethanol yield were limited by saccharification at 10 and 20 filter papaer units (FPU) g-1 acid-treated bagasse. Dilute slurries of biomass were converted to ethanol more efficiently (over 72% of theoretical yield) in simple batch fermentations than slurries containing high solids albeit with the production of lower levels of ethanol. With high solids (i.e., 160 g acid-treated bagasse L-1), a combination of 20 FPU cellulase g-1 bagasse, preincubation under saccharification conditions, and additional grinding (to reduce particle size) were required to produce ca. 40 g ethanol L-1. Alternatively, almost 40 g ethanol L-1 was produced with 10 FPU cellulase g-1 bagasse by incorporating a second saccharification step (no further enzyme addition) followed by a second inoculation and short fermentation. In this way, a theoretical ethanol yield of over 70% was achieved with the production of 20 g ethanol 800 FPU-1 of commercial cellulase. © 1994 John Wiley & Sons, Inc.
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  • 21
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    Biotechnology and Bioengineering 44 (1994), S. 329-336 
    ISSN: 0006-3592
    Keywords: biofilm formation ; Escherichia coli ; C/N ratio ; plasmid retention ; extracellular polysaccharide ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Biofilm formation and plasmid segregational instability in biofilm cultures of Escherichia coli DH5α (pMJR1750) were investigated under different medium-carbon-to-nitrogen (C/N) ratios. At C/N ratios of 0.07 and 1, net accumulation of both biofilm plasmid-bearing and plasmid-free cells continued through the entire experiment without attaining any apparent steady state. At C/N ratios of 5 and 10, net biofilm cell accumulation for the two populations reached apparent steady states after 84 and 72 h, respectively. At C/N ratios of 0.07 and 1, polysaccharide production increased slowly and reached about 2g alginate equivalent/cm2 by the end of both experiments. At a C/N ratio of 5, polysaccharide increase significantly after 84 h, reaching about 7μg alginate equivalent/cm2 prior to termination. At a C/N ratio of 10, polysaccharide increased significantly after 72 h and reached 21 μg alginate equivalent/cm2 at 108 h. At C/N ratios of 0.07 and 1, protein production reached 6.5 and 4 μg/cm2, respectively. At C/N ratios of 5 and 10, protein production increased slightly for the first 84 h and reached a maximum at 108 h, at 3 and 2 μg/cm2, respectively, then decreased over the last 12 h of the experiment. Ratios of polysaccharide to protein increased with increasing C/N ratios. At C/N ratios of 0.07 and 1, the ratios between extracellular polysaccharide (EP) and protein were no more than 205 μg polysaccharide/μg protein, whereas those at C/N ratios of 5 and 10 increased to about 7 and 12 μg polysaccharide/μg protein, respectively.Probabilities of plasmid loss in the biofilm cultures increased with increasing C/N ratios. At C/N ratios of 0.07, 1, and 5, the probabilities of plasmid loss were 0.0013 ± 0.011, 0.020 ± 0.006 and 0.122 ± 0.021, respectively. At a C/N ratio of 10, the probability of plasmid loss was significantly higher, reaching 0.38 ± 0.125. The increase of probability of plasmid loss at higher C/N ratios results from competition between cell replication and extracellular polysaccharide production. © 1994 John Wiley & Sons, Inc.
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  • 22
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    Biotechnology and Bioengineering 44 (1994), S. 165-169 
    ISSN: 0006-3592
    Keywords: penicillin ; fermentation ; media ; degradation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In Industrial production of penicillin there is a noticeable loss of the product through degradation reactions. It is shown that the degradation of penicillin-V, both in a complex and in a chemically defined medium, can be separated into a phosphate-catalyzed conversion of penicillin-V to penicilloic-V acid, overlaid by at least one other reaction in which penicillin V is degraded to as yet unknown products. Parameter values for the phosphatecatalyzed degradation are found to be independent of the type of fermentation medium. The rate of formation of other degradation products of penicillin-V is found to be significantly higher in a complex fermentation medium with corn-steep liquor in a chemically defined medium. © 1994 John Wiley & Sons, Inc.
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  • 23
    ISSN: 0006-3592
    Keywords: biosensor ; ethanol ; fermentation ; lignocellulose hydrolysate ; liquid chromatography ; microdialysis ; on-line ; sampling ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: During a 70-h fermentation of a lignocellulose hydrolysate, the ethanol produced was monitored on-line using a microdialysis probe as an in situ sampling device. The dialysate components were then separated in a column liquid chromatographic system and the ethanol was selectively detected by an amperometric alcohol biosensor. The result was compared with two off-line analysis methods: one chromatographic method with refractive index (RI) detection and one enzymatic method based on spectrophotometric detection. The two methods base on enzymes were shown to give lower values than the chromatographic method based on RI detection, which is discussed n terms of selectivity. The investigated on-line setup was found to be a flexible system for monitoring of fermentations, allowing a sampling frequency of at least 12 h-1 and with a delay between sampling and detection of less than 5 min. © 1994 John Wiley & Sons, Inc.
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  • 24
    ISSN: 0006-3592
    Keywords: hybridoma ; fed batch ; materials balancing ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: Hybridoma batch cultures were extended using feed formulations based on nutrient consumption measured during different batch culture phases when (a) growth but negligible antibody production was taking place; (b) maximum antibody production rate and declining viable cell growth rate were observed. Strategy (a) was the more successful (2.8-fold compared with 1.8-fold antibody titer increase) and maintained cell viability for longer. Analysis of the effects of omitting individual amino acids yielded results which were consistent with those from the feeding experiment © 1994 John Wiley & Sons, Inc.
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  • 25
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    Biotechnology and Bioengineering 43 (1994), S. 847-855 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; cellular energetics ; acetate production ; carbon yield ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An integrated metabolic model for the production of acetate by Escherichia coli growing on glucose under aerobic conditions was presented previously (Ko et al., 1993). The resulting model equations can be used to explain phenomena often observed with industrial fermentations, i.e., increased acetate production which follows from high glucose uptake rate, a low dissolved oxygen concentration, a high specific growth rate, or a combination of these conditions. However, several questions still need to be addressed. First, cell composition is growth rate and media dependent. Second, the macromolecular composition varied between E. coli strains. And finally, a model that represents the carbon fluxes between the Embden-Meyerhof-Parnas (EMP) and the hexose monophosphate (HMP) pathways when cells are subject to internal and/or external stresses is still not well defined. In the present work, we have made an effort to account for these effects, and the resulting model equations show good agreement for wild-type and recombinant E. coli experimental data for the acetate concentration, the onset of acetate secretion, and cell yield based on glucose. These results are useful for optimizing aerobic E. coli fermentation processes. More specifically, we have determined the EMP pathway carbon flux profiles required by the integrated metabolic model for an accurate fit of the acetic acid profile data from a wild-type E. coli strain ML308. These EMP carbon flux profiles were correlated with a dimensionless measurement of biomass and then used to predict the acetic acid profiles for E. coli strain F-122 expressing human immunodeficiency virus-(HIV528) β-galactosidase fusion protein. The effect of different macromolecular compositions and growth rates between these two E. coli strains required a constant scaling factor for improved quantitative predictions.
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    Biotechnology and Bioengineering 44 (1994), S. 21-28 
    ISSN: 0006-3592
    Keywords: cell recycle ; thermophilic ethanol fermentation ; Bacillus ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Continuous fermentation with cell recycle proved very effective in increasing the ethanol volumetric productivity of the thermophilic facultative anaerobe, Bacillus stearothermophilus strain LLD-15, on sucrose at 70°C. When complete cell recycle was used, cell viability decreased after a few residence times and sucrose consumption was reduced. Operation using a constant bleed rate resulted in greater stability and higher ethanol volumetric productivities. A mathematical model based on maintenance energy requirements provided an adequate description of the system. © 1994 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 44 (1994), S. 87-94 
    ISSN: 0006-3592
    Keywords: glutamic acid ; fermentation ; fuzzy control system ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: In glutamic acid fermentation, the molasses feeding policy and time of penicillin addition significantly affected glutamic acid production, and a fuzzy supervisory control system was developed for their quasi-optimal regulation.From the trend of the experimental data, production rules and membership functions of fuzzy inference were devised to determine the quasi-optimum molasses feeding policy and penicillin addition time. A computer with multitasking operating system was used for the construction of the control system with fuzzy inferencing, which decided the control policy every minute, and the feed rate was controlled automatically. The pattern of residual sugar concentration was almost the same as that of maximum glutamic acid production under manual operation. Using the computer control system, stable production was maintained at the highest level of 71 to 75 g/L. © 1994 John Wiley & Sons, Inc.
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  • 28
    ISSN: 0006-3592
    Keywords: acetate reduction ; Bacillus subtilis ; Escherichia coli ; cloning ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: A novel metabolic engineering technique involving the redirection ofcellular carbon fluxes was employed to reduce acetate production in an Escherichia coli culture. Metabolic engineering was achieved by cloning E. coli the gene for the Bacillus subtilis acetolactate synthase (ALS), an enzyme capable of catalyzing the conversion of pyruvate to nonacidic and less harmful species. The heterologous expression of the ALS catabolic enzyme in Escherichia coli drastically modified the cellular glycolytic fluxes. In particular, acetate excretion, which is a common characteristic of E. coli, as well as a physiological burden, was minimized. The residual acetate level was kept under control and maintained at a level that was below the toxic threshold. The expression of the biologically active ALS enzyme in E. coli did not result in any detectable changes on either cell growth rate or cell yields. The alternative product, acetoin, was shown to be 50 times less harmful than acetate. Similarities in the growth pattern of two different E. coli strains, RR1 and GJT001, under all cultivation conditions suggested that the ability of ALS to reduce acetate accumulation is generic and not strain-specific. © 1994 John Wiley & Sons, Inc.
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  • 29
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    Biotechnology and Bioengineering 44 (1994), S. 952-960 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; protein production, recombinant ; glucose uptake ; acetate excretion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Reduction of acetate excretion using a modified cellular glucose uptake rate was examined. An Escherichia coli strain bearing a mutationin ptsG, a gene encoding enzyme II in glucose phosphotransferase system (PTS), was constructed and characterized. The growth rate of the mutant strain was slower than its parent in glucose defined medium, butwas not affected in complex medium. Experimental results using this mutant strain showed a significant improvement in culture performance in simple batch cultivations due to reduced acetate excretion through the modified glucose uptake. Both biomass and recombinant protein productivity were increased by more than 50% with the ptsG mutant when compared to the parent strain. Recombinant protein productivity by the newly constructed strain at a level of more than 1.6 g/L was attained consistently in a simple batch bioreactor. © 1994 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 43 (1994), S. 969-977 
    ISSN: 0006-3592
    Keywords: cross-flow membrane filtration ; inclusion bodies ; Escherichia coli ; extraction, rIL-2 ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A cross-flow membrane filtration process was developed for the recovery of rIL-2 inclusion bodies from homogenized Escherichia coli. The membrane extraction process was comprised of a two-step diafiltration followed by an extraction with 7 M GuHCl and a 40-fold dilution of the solubilized inclusion bodies into 0.01 M Tris-HCl, 0.035 M NaCl, pH 7.9. The first diafiltration was with a 0.03 M Tris-HCl, 5 mM ethylenediaminetetraacetic acid (EDTA), pH 8, followed by a diafiltration with 1.75 M GuHCl. All of the insoluble rIL-2 was retained behind the membrane, whereas a GuHCl wash solubilized approximately 15% of the rIL-2. The membrane process increased the yield of rIL-2 in the diluted extract by threefold as compared to a similar centrifuge process with a significant increase in purity as determined by reverse-phase high-performance liquid chromatography (HPLC). © 1994 John Wiley & Sons, Inc.
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  • 31
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    Journal of industrial microbiology and biotechnology 12 (1993), S. 256-262 
    ISSN: 1476-5535
    Keywords: Listeria ; Salmonella ; Shigella ; Aeromonas ; Staphylococcus ; Escherichia coli ; Bacillus cereus ; Clostridium botulinum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary MKES Tools is a microbial kinetics expert system for developing food production systems and assessing product safety. The specific information required as input are: (1) a flowchart of the production system, (2) the factors affecting the survival and growth of food-borne pathogens and (3) the ranges of variation for each factor's parameters. With this information, MKES Tools simulates the growth and survival of pathogenic microorganisms when subjected to many different factor/parameter situations. The responses obtained are then used to estimate the significance of each factor's parameters.
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  • 32
    ISSN: 1476-5535
    Keywords: Gene transfer ; Escherichia coli ; River water ; Indigenous bacteria
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary This study examined the transfer of the plasmid pBGH1, an expression vector for bovine somatotropin (BST), fromEscherichia coli K-12 strain W3110G [pBGH1] to indigenous microorganisms present in flasks containing Missouri River water. Strain LBB269 is a nalidixic acid-resistant derivative of W3110G which was used as a plasmid-free control strain in these studies. Water samples were inoculated with strains W3110G [pBGH1] and LBB269; after 21 days of incubation the number of viable colony-forming units (CFU) of W3110G [pBGH1] and LBB269 were reduced from an initial level of about 1×107 CFU per ml to less than 1 CFU per 100 ml. At this time indigenous microbes resistant to both ampicillin and tetracycline (the antibiotic resistance markers on pBGH1) were isolated from 100 ml of water from each of the flasks inoculated with either strain W3110G [pBGH1] or LBB269. Plasmid DNA was isolated from these organisms and examined for sequences containing the gene for BST from pBGH1, using a polymerase chain reaction (PCR) assay. As expected, the day 0 sample from the flask inoculated withE. coli K-12 strain W3110G [pBGH1] gave a positive PCR response and the day 0 sample from the flask inoculated withE. coli K-12 strain LBB269 gave a negative PCR response. All of the day 21 samples containing indigenous microbes isolated from flasks that were inoculated with either W3110G [pBGH1] or LBB269 were negative in the PCR assay, indicating that the target sequence from pBGH1 was not present in any of these indigenous microorganisms. The results of this particular assay indicate that pBGH1 or the portion of pBGH1 including the BST structural gene had not been transferred from W3110G [pBGH1] to indigenous microbial inhabitants of the Missouri River water flasks during this study.
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  • 33
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    Journal of industrial microbiology and biotechnology 11 (1993), S. 253-257 
    ISSN: 1476-5535
    Keywords: Colonization ; Escherichia coli ; Gastrointestinal ; Environmental ; Survival
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The colonizing potential ofEscherichia coli K12 containing a vector coding for somidobove (bovine somatotropin) was determined. Treated male and female Fischer-344 rats were given a single oral gavage inoculum of sucrose with/without tetracycline (15 μg/ml). Untreated control animals received similar drinking water regimes. All animals survived until termination. There were no clinical signs of toxicity observed and no treatment-related effect upon body weight, food consumption, or efficiency of food utilization. Fresh fecal samples were collected from each rat every 24 h following inoculation and the population of the marked strain was quantitated until no bacterial colonies were observed for two consecutive days. While all inoculated rats were positive at 24 h, by 72 and 96 h all had become negative for the test (marked) strain, as were the corresponding control group throughout the test. The frozen stock of the marked strain used as the positive control demonstrated that the agar plates were selective for the test strain. Fourteen days following inoculation, all groups of rats were killed and the gastrointestinal tracts removed and treated to recover the marked strain. There was no evidence of the marked strain in the gastrointestinal tract of any rat from any group. Thus, theE. coli K12 host/vector system used in this experiment does not colonize the gastrointestinal tract of Fischer-344 rats.
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    World journal of microbiology and biotechnology 9 (1993), S. 34-36 
    ISSN: 1573-0972
    Keywords: Bacteriophages ; Escherichia coli ; Salmonella ; sewage ; wastewater
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A new method for quantifying F-specific bacteriophages in wastewater is described. Somatic coliphages were also determined. Host-strainSalmonella typhimurium WG 49 was sensitive to only a few bacteriophages and this could have arisen from infection by F-RNA phages. Host-strainEscherichia coli ATCC 9723 C, however, supported multiplication of a wide range of bacteriophages present in sewage, giving plaque counts one to three orders of magnitude greater than those on F+ and F- salmonellas.
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  • 35
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    World journal of microbiology and biotechnology 9 (1993), S. 269-274 
    ISSN: 1573-0972
    Keywords: Enterobacteriaceae ; fermentation ; lactic acid bacteria ; maize ; nixtamal ; pozol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Freshly prepared pozol, a traditional Mexican fermented maize dough, contained (c.f.u./g wet wt): lactic acid bacteria, 104 to 106; aerobic mesophiles, 104 to 105; Enterobacteriaceae, 102 to 103; yeasts, 102 to 104; and mould propagules, 〈103. After 30 h at 28°C the numbers were, respectively: 109, 7×106, 5×105, 106 and 104. Soaking alkali-treated grains overnight allowed lactic acid bacteria, aerobic mesophiles and Enterobacteriaceae to grow and these then constituted the primary microbial flora of the pozol dough. Grinding in a commercial mill inoculated the dough with lactic acid bacteria, aerobic mesophiles, Enterobacteriaceae and yeasts. Other processing stages, including the nature of the surface upon which the balls were made, handling of the dough, and air, contributed only minor numbers of microbes compared with the two major sources, soaking and grinding. The pH of pozol fell from an initial value of 7.3 to 4.6 after 30 h incubation at 28°C. The numbers of Enterobacteriaceae and other aerobic mesophilic bacteria remained constant between 11 and 30 h incubation and there was no evidence of the acidic conditions having any lethal effects on these organisms.
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  • 36
    ISSN: 1573-0972
    Keywords: Ethanol ; fermentation ; modelling ; molasses ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Batch fermentations of sugar-cane blackstrap molasses to ethanol, using pressed yeast as inoculum, demonstrated an exponential relationship between the time necessary to complete the fermentation and the initial concentrations of sugar and yeast cells. The parameters of the derived exponential equations depended on the experimental conditions.
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    World journal of microbiology and biotechnology 9 (1993), S. 313-318 
    ISSN: 1573-0972
    Keywords: Cellulose ; eucalyptus wood ; fermentation ; kinetic model ; saccharification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Coupled saccharification and fermentation of Eucalyptus globulus wood, pre-treated by acid hydrolysis and sodium hypochlorite, was carried out in two column reactors: one for enzymatic hydrolysis of the substrate at 50°C and the other for fermentation of sugars with calcium alginate-immobilized Saccharomyces cerevisiae at 30°C. A buffered solution containing cellulases at pH 4.8 was recycled through both reactors. The maximum yields were about 0.26 g ethanol per g of substrate. The results were reproduced reasonably well using a simple kinetic model consisting of two successive pseudo-first-order reactions.
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  • 38
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    World journal of microbiology and biotechnology 9 (1993), S. 603-604 
    ISSN: 1573-0972
    Keywords: Cocoa beans ; fermentation ; microbial populations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Acetic and lactic acid bacteria on fermented cocoa beans were maximally 2.0×106 and 1.9×106 c.f.u./g wet wt, respectively. Acetic and lactic acids were detected on the second and fourth days of fermentation and were maximally 140 and 45 mg/10 g beans, respectively. There was a positive correlation between the sizes of the relevant microbial populations and the amounts of acids produced during fermentation.
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  • 39
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    World journal of microbiology and biotechnology 9 (1993), S. 609-610 
    ISSN: 1573-0972
    Keywords: Antibiotics ; Escherichia coli ; resistance ; river water
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Of 107 Escherichia coli strains isolated from the water, sediment and fish of the Bhavani River, all of which are considered potential causes of human enteric disease, 62% were resistant to more than four antibiotics. Levels of resistance to bacitracin, penicillin, and novobiocin were generally high whereas those to polymyxin-B and chloramphenicol were much lower. A high incidence of multiple antibiotic resistant E. coli was noted in all samples and the multiple antibiotic resistance index of the strains showed that 95% of the strains originated either from man or cattle.
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    World journal of microbiology and biotechnology 9 (1993), S. 295-299 
    ISSN: 1573-0972
    Keywords: Bacillus ; Enterococcus ; fermentation ; kinema ; soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Kinema fermentations of Indian and Canadian No. 1 soybeans by Bacillus sp. DK-W1 and by mixed cultures of Bacillus sp. DK-W1 and Enterococcus faecium DK-C1 were essentially identical. The viable cell count of Bacillus increased from an initial 105 to 1010 c.f.u./g wet wt after 48 h incubation at 37°C. The pH of the fermentation dropped from an initial 6.9 to about 6.4 after 8 h and then rose to 8.6 after 32 h, with a coincident increase in proteolytic activity and ammonia concentration. The fermentations containing E. faecium and Bacillus exhibited a greater initial pH decline and a slightly retarded subseqent increase in pH compared with fermentations with Bacillus only. The presence of E. faecium had no detectable effects on growth of the Bacillus, proteolytic activity, ammonia production or the final pH of the fermentations.
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    World journal of microbiology and biotechnology 9 (1993), S. 381-382 
    ISSN: 1573-0972
    Keywords: Cocoa beans ; fermentation ; products
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The maximum amounts of acetic acid produced by ripe and unripe cocoa beans were 157 mg and 110 mg/10 g wet wt of cotyledon, respectively. The unripe beans had a lower pH than the ripe beans after 6 days' fermentation. About 40% of ripe beans achieved a chocolate colour compared with 27% of unripe beans.
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  • 42
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    Biotechnology and Bioengineering 41 (1993), S. 316-324 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; acetic acid ; inhibition ; glycine ; methionine ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Among amino acids screened for their potential to relieve wild and recombinant Escherichia coli from the negative effects of acetic acid, glycine, and methionine showed a sparing effect. In the presence of 2 g/L of acetic acid, addition of 0.5 g/L of glycine or methionine resulted in either a complete recovery or a further enhancement in the specific growth rate, while the enhancement was significant but not fully complete in the presence of 4 g/L of acetic acid. The addition of 0.5 g/L of methionine alleviated the negative effect of acetic acid on recombinant E. Coli growth to produce more β-lactamase, which was encoded by plasmid pUC18. In continuous fermentation the methionine effect on recombinant. E. coli metabolism depended on dilution rate; at high dilution rates, above 0.4 h-1, the methionine addition enhanced β-lactamase production and reduced acetic acid formation, while at low dilution rates, below 0.3 h -1, the effect was reversed. In def-batch fermentation with wild-type E. Coli, cell growth rate and cell yield from glucose were enhanced with methionine addition, while the acetic acid concentration reached over 4 g/L. © 1993 John Wiley & Sons, Inc.
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  • 43
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    Biotechnology and Bioengineering 42 (1993), S. 30-36 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; fiber optic ; firefly luciferase ; on-line ; viability ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel method is described for the on-line determination of viable cell number. It has been tested in fermentations of Escherichia coli. The cells are transfected with the gene for firefly luciferase and fed low levels of luciferin in the medium. The reaction requires ATP, so the nonviable cells cannot produce light. Thus, light production is linear with viable cell density from innoculation through most of exponential growth. The light emitted by these cells is then conducted from the reaction vessel to the light detection equipment by an optical fiber. With the equipment described below, as few as a 106 cells/mL, or an OD600 of 0.004, are easily detectable and concentrations greater than 1010 cells/mL are well within range. The data are collected by a computer, so adaptation to on-line control applications is straightforward. During lag phase, this method is much more accurate then optical density measurements. At the end of exponential growth, rapid changes in light production mark carbon source depletion and the onset of cell lysis. A simple model accounts for the luciferin used during the fermentation and corrects the light detected to the proper cell density. © 1993 John Wiley & Sons, Inc.
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  • 44
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    Biotechnology and Bioengineering 42 (1993), S. 59-73 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; amino acids ; nucleotides ; biosynthesis ; linear optimization ; metabolic fluxes ; metabolic engineering ; stoichiometry ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Microbial metabolism provides at mechanism for the conversion of substrates into useful biochemicals. Utilization of microbes in industrial processes requires a modification of their natural metabolism in order to increase the efficiency of the desired conversion. Redirection of metabolic fluxes forms the basis of the newly defined field of metabolic engineering. In this study we use a flux balance based approach to study the biosynthesis of the 20 amino acids and 4 nucleotides as biochemical products. These amino acids and nucleotides are primary products of biosynthesis as well as important industrial products and precursors for the production of other biochemicals. The biosynthetic reactions of the bacterium Escherichia coli have been formulated into a metabolic network, and growth has been defined as a balanced drain on the metabolite pools corresponding to the cellular composition. Theoretical limits on the conversion of glucose, glycerol, and acetate substrates to biomass as well as the biochemical products have been computed. The substrate that results in the maximal carbon conversion to a particular product is identified. Criteria have been developed to identify metabolic constraints in the optimal solutions. The constraints of stoichiometry, energy, and redox have been determined in the conversions of glucose, glycerol, and acetate substrates into the biochemicals. Flux distributions corresponding to the maximal production of the biochemicals are presented. The goals of metabolic engineering are the optimal redirection of fluxes from generating biomass toward producing the desired biochemical. Optimal biomass generation is shown to decrease in a piecewise linear manner with increasing product formation. In some cases, synergy is observed between biochemical production and growth, leading to an increased overall carbon conversion. Balanced growth and product formation are important in a bioprocess, particularly for nonsecreted products. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 42 (1993), S. 50-58 
    ISSN: 0006-3592
    Keywords: emulsion liquid membrane ; lactic acid ; organic acid recovery ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Studies on the batch extraction of lactic acid using an emulsion liquid membrane system are reported. The membrane phase consists of the tertiary amine carrier Alamine 336 and the surfactant Span 80 dissolved in n-heptane/paraffin and aqueous solutions of sodium carbonate in the internal phase. The effects of internal phase reagent, extraction temperature, and initial external phase pH on the extraction efficiency and the emulsion swelling are examined. A statistical factorial experiment on extraction from clarified lactic acid fermentation broth was carried out to obtain knowledge of the performance of the extraction system from a broth. The extraction efficiency from the fermentation broth is found to be lower as compared to aqueous solutions of pure lactic acid. The effect of pH and the presence of other ionic species on selectivity are discussed. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 221-230 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; medium optimization ; chemostat ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An optimized, defined minimal medium was developed to support balanced growth of Escherichia coli X90 harboring a recombinant plasmid. Foreign protein expression was repressed in these studies. A pulse injection technique was used to identify the growth responses to nutrients in a chemostat. Once the nutrients essential for growth had been identified, the yield coefficients for individual medium components. These yield coefficients were used to develop an optimized, glucose-limited defined minimal medium that supports balanced cell growth in chemostat culture. The biomass and substrate concentrations follow the Monod chemostat model. The maximum specific growth rate determined in a washout experiment is 0.87 h-1 for this strain in the optimized medium. the glucose yield factor is 0.42 g DCW/g glucose and the maintenance coefficient is zero in the glucose-limited chemostat culture. © 1993 John Wiley & Sons, Inc.
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  • 47
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    Biotechnology and Bioengineering 41 (1993), S. 830-832 
    ISSN: 0006-3592
    Keywords: ionic equilibrium ; pH computation ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An algorithm developed for pH computation has been tested to calculate the theoretical pH changes in a culture medium during the course of a fermentation. A divergence between the computed pH value and the value measured with the electrode allows us to highlight the presence of undetected ionic products. The calculation with the algorithm by means of a computer requires only the knowledge of the ionic properties of the substrates and detected products and existing thermodynamic constants. © 1993 Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 3-13 
    ISSN: 0006-3592
    Keywords: recombinant protein ; Escherichia coli ; inclusion body ; renaturation ; disulfide bond ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Expression of recombinant proteins in Escherichia coli often results in the formation of insoluble inclusion bodies, In case of expression of eukaryotic proteins containing cysteine, which may form disulfide bonds in the native active protein, often nonnative inter- and intramolecular disulfide bonds exist in the inclusion bodies. Hence, several methods have been developed to isolate recombinant eukaryotic polypeptides from inclusion bodies, and to generate native disulfide bonds, to get active proteins. This article summarizes the different steps and methods of isolation and renaturation of eukaryotic proteins containing disulfide bonds, which have been expressed in E. coli as inclusion bodies, and shows which methods originally developed for studying the folding mechanism of naturally occurring proteins have been successfully adapted for reactivation of recombinant eukaryotic proteins. © 1993 John Wiley & Sons, Inc.
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  • 49
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    Biotechnology and Bioengineering 41 (1993), S. 237-244 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; iron transport ; enterobactin HPLC ; dialysis membrane fermentor ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The article describes four different fermentation procedures for Escherichia coli AN311, a producer of enterobactin. A regular rotary shaker culture with a biphasic system consisting of an agar layer (as a reservoir for feeding processes) and a layer of liquid medium, 2.4 L and 10 L batch cultures, and a novel dialysis membrane fermentor were used. With the use of this latter fermentor type, the production of enterobactin could be increased by a factor of about 9.5, while growth increased by a factor of 12 compared to the other systems. For the rapid and reliable quantification of the concentration and purity of enterobactin an analytical and preparative high-performance liquid chromatography (HPLC) method was established. The degradation compounds of this siderophore were detected by diodearray and bioassays. A comparison of total catechol production as well as the distribution between enterobactin and its degradation compounds is given. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 361-369 
    ISSN: 0006-3592
    Keywords: gravitational sedimentation ; sedimentation ; fermentation ; continuous ; Saccharomyces cerevisiae ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A mathematical model for the sedimentation velocity in an inclined parallel plate sedimenter is proposed. The parameters of the alcoholic fermentation broth (cell density of Saccharomyces cerevisiae, density of the fermentation medium, viscosity of the broth at various alcohol and biomass contents) were determined experimentally. The sedimentation velocities were predicted under the various operational conditions and parameters, both of the broth (the alcohol concentration and cell content) and the sedimenter prototype (length, distance between the plates, and slope). The proposed model for the sedimentation velocity presented a good correlation with the experimental results of continuous sedimentation. These sedimenter prototypes were assembled and tested for efficiency of separation of yeast cell under conditions considered for interest for continuous alcoholic fermentation. A selective filter for the overflow composed of calcium alginate gel improved operation. A high operational stability, high separation efficiency (over 98%), and adequate settler residence times (about 20 min) were attained. The operational results permitted the operation of continuous alcoholic fermentation with cellular recycling effected exclusively by gravitational sedimentation, this characterizing a process of enormous industrial interest because of the operational simplicity and low operational and maintenance costs. © 1993 John Wiley & Sons, Inc.
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  • 51
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    Biotechnology and Bioengineering 41 (1993), S. 395-404 
    ISSN: 0006-3592
    Keywords: recombinant bacterium ; plasmid loss ; modeling ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A large number of models concerning cultures of genetically engineered bacteria have been described. Among them, some are specifically adapted to continuous cultures and lead to the determination of two variables: (i) the difference in the specific growth rates between plasmid-carrying cell and plasmid-free cells (δμ) and (ii) the frequency of plasmid loss by plasmid-containing cells (prμ+). Until now, studies have been performed on the global expression prμ+ and δμ, whose value during continuous assays have been supposed approximately constant (mean value) and not on separate values of both terms pr and μ+, respectively, probability of plasmid loss and specific growth rate of the plasmid-carrying cells. So far these studies do not allow examination of the relationship between these two last parameters. Experimental results were obtained with Escherichia coli C600 galk (GAPDH), a genetically engineered strain that synthetizes an elevated quantity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). From data obtained during continuous cultures, it is shown that during an assay, δμ, and prμ+ do not remain constant. An appropriate mathematical analysis of the expression of μ- (specific growth rate of the plasmid-free cells) and μ+ has been built up. This allows the evaluation of the values of μ+ and μ- during the continuous cultures carried out at different dilution rates. Values of pr have been calculated from these data. Indeed our results show that pr increases with μ+. A modeling approach which allows correct simulation of this variation is also proposed. This model is derived from the Hill equation regarding cooperative binding of enzymic type reaction. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 503-511 
    ISSN: 0006-3592
    Keywords: mathematical modeling ; fermentation ; Claviceps purpurea ; alkaloid production ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An new systematic approach for describing Claviceps purpurea growth and ergot alkaloid production during batch fermentation is presented. The model is based on microbial life, as the main characteristic for microbial development during fermentation process. The aging process of the microorganism is represented by life function, defined in microbial life space. The life space is defined as a measure in which the observer follows the development of a biosystem through physiological and morphological changes of a microorganism. As a consequence of such approach the relativistic theory is recognized. To validate the model developed, a test on growth and alkaloid synthesis data from an industrial batch fermentation was performed. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 666-670 
    ISSN: 0006-3592
    Keywords: oxygen fluctuations ; plasmid amplification ; Escherichia coli ; circulation time distribution ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Escherichia coli DH5α, carrying the pUC19 plasmid for the lacZ fragment of β-galactosidase and ampicillin resistance, was grown in a batch fermentor under conditions of fluctuating oxygen supply. A Monte Carlo method was used to control the on/off supply of air to simulate circulation of cells in a large fermentor. Rapid changes in oxygen supply reduced the rates of oxygen uptake the carbon dioxide release and prolonged the active second growth phase in batch culture, compared to growth with continuous aeration. Amplification of the plasmid was observed during the stationary phase when air supplied continuously, but not during the Monte Carlo experiments. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 937-946 
    ISSN: 0006-3592
    Keywords: protein excretion ; continuous culture ; Escherichia coli ; β-lactamase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The stable continuous overproduction of a plasmidencoded protein, β-lactamase, for at least 50 days by Escherichia coli K-12, RB791(pKN), with release into the culture medium has been demonstrated in two-stage chemostats. The second-stage culture was continuously induced with 0.1 mM IPTG. Continuous expression of β-lactamase could not be sustained with this strain in a single-stage chemostat because of cell death and selection for lac-1 cells. β-Lactamase production in the second stage was sensitive to the second-stage dilution rate and the distribution of the limiting substrate (i.e., glucose) between the first and second stages. The fraction of viable, excreting cells and the average copy number in the induced culture was measurably higher under those conditions of dilution rate and substrate distribution which yielded high β-lactamase levels. The best operating conditions found at 20°C were a first-stage dilution rate of 0.12 h-1, a second-stage dilution rate of 0.03 h-1, and equal glucose feed supplied to each stage. Enzymatically active β-lactamase was produced at a level of 25% of total cellular protein with 90% excretion yielding 300 mg β-lactamase/L that was 50% pure at an OD600 〈 6. © 1993 Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 42 (1993), S. 215-221 
    ISSN: 0006-3592
    Keywords: on-line NMR ; phosphorus-31 NMR ; Escherichia coli ; aerobic and anaerobic metabolism ; intracellular pH ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An experimental system has been constructed which enables on-line measurements of phosphorus-31 (31P) nuclear magnetic resonance (NMR) spectra for growing bacterial suspensions under anaerobic or aerobic conditions. A sample stream from a laboratory bioreactor is circulated to the NMR sample chamber in a gas exchange system which permits maintenance of aerobic conditions for high-cell-density cultures. 31P NMR spectra with resolution comparable with those obtained traditionally using dense, concentrated, nongrowing cell suspensions can be obtained at cell densities above 25 g/L with acquisition times ranging from 14 to 3 minutes which decline as cell density increases. This system has been employed to characterize the changes in intracellular state of a stationary phase culture which is subjected to a transition from aerobic to anaerobic conditions. Both intracellular NTP level and cytoplasmic pH are substantially lower under anaerobic conditions. Also, the system has been employed to observe the response of a growing culture to external addition of acetate. Cells are able to maintain pH difference across the cytoplasmic membrane at extracellular acetate concentrations of 5 and 10 g/L. However, acetate concentrations of 20 g/L cause collapse of the transmembrane ΔpH and sharp reduction of the growth rate of the culture. The experimental configuration described should also permit NMR observations of many other types of microbial cultures and of other nuclei. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 42 (1993), S. 538-541 
    ISSN: 0006-3592
    Keywords: ethanol ; fermentation ; levoglucosan ; lignocellulose ; pyrolysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The Waterloo Fast Pyrolysis Process (WFPP) can produce an organic liquid high in levoglucosan (1, 6-anhydro-β-D-glucopyranose) content from suitably pretreated lignocellulosics. A variety of fungi and yeasts were screened for their ability to utilize and ferment this organic liquid. To enhance its fermentability, the pyrolysis tar was posttreated in three different ways: (1) an aqueous extract (lignin removed); (2) activated charcoal treated (lignin and aromatics removed); and (3) acid hydrolysate (lignin and aromatics removed with the levoglucosan hydrolyzed to glucose). Four fungal strains were examined. None grew in the aqueous extract, but all grew equally well in both the activated charcoal treated and the acid hydrolysate, suggesting that the aromatic species were inhibitory to growth. Seven yeast species were examined, two of which did not grow on any of the extracts. Five of the yeast strains grew well on both the aqueous extract as well as the activated charcoal extract. The hydrolysate was optimal in terms of biomass yield and ethanol production. Ethanol yields on the hydrolysate were comparable or better than those on glucose. Ethanol was also produced in the aqueous extract and activated charcoal-treated substrate, but yields were considerably lower than on the hydrolysate or glucose. It is apparent that a wood pyrolysate maximized for levoglucosan can serve as a fermentable substrate, although postpyrolysis clean-up appears necessary. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 42 (1993), S. 1175-1180 
    ISSN: 0006-3592
    Keywords: self-cycling fermentation ; secondary metabolite ; biosurfactant ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Self-cycling fermentations (SCFs) were conducted in a stirred tank apparatus using Bacillus subtilis and Acinetobacter calcoaceticus. The systems were very stable and the experiments lasted through many cycles. The variation of parameters such as biomass and doubling time from cycle to cycle was small. The stirred tank reactor (STR) allowed a much better control of the working volume in the fermentor from cycle to cycle, compared to the cyclone column, and it was not necessary to make periodic corrections.The production of surfactin from B. subtilis was achieved without extending the cycle time. The harvested broth at the end of each cycle was allowed to remain in a secondary vessel, at ambient temperature, before being collected. It is exhaustion of the limiting nutrient which causes an increase in dissolved oxygen (DO). At this point, the computer, which constantly monitors the DO, triggered the harvesting sequence to end the cycle. Thus, the mature culture in the secondary vessel experienced appropriate conditions for the production of the secondary metabolite. Meanwhile, the next batch of cells was being grown in the primary reactor.The response of a gas analyzer on the effluent paralleled that of the DO measurements in the fermentor. These data for oxygen and carbon dioxide exhibited less noise than the DO readings. Either would be a more reliable parameter for feedback control of the SCF because the problem of fouling of the DO probe after extended runs of many cycles would be eliminated. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 148-155 
    ISSN: 0006-3592
    Keywords: plasmid stability ; Streptomyces lividans ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Plasmid plJ303 stability in Streptomyces lividans cultures has been studied by measuring plasmid copy number under various growth conditions. An increase in mean plasmid copy number was normally seen during early rapid growth in both shaken culture and stirred vessel fermentations at 28°C. Maximum copy numbers were consistently attained in early stationary phase followed by a decline (of variable amount) upon further incubation. The imposition of environmental stress (high growth temperature, i.e., 37°C, and low dissolved oxygen tension, i.e., 〈5% air saturation) led to a plasmid copy number of zero and a 50% reduction, respectively. Interestingly, the relative proportions of plasmid topoisomers changed with time since progressively more supercoiled forms were observed throughout the stationary phase. Plasmid dimers were also observed in some cultures, and no evidence of structural plasmid instability was found. In general, this host-vector system seemed remarkably stable under normal growth conditions. However, copious organic acid production by the host was observed and was thought to be undesirable for good heterologous gene expression of a secreted protein. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 325-329 
    ISSN: 0006-3592
    Keywords: chemostat ; enzyme overproduction ; plasmid stability ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of plasmid-mediated metabolic burden of on the expression of the host genes and its consequences on the plasmid maintenance were studied in carbon-limited chemostat culture of Escherichia coli 1EA(pBR322) subject to selection for strains overproducing chromosomally coded ribitol dehydrogenase. The chemostat population became rapidly heterogeneous and the competition among evolved strains was found to be crucial for the kinetics of the plasmid loss from the culture. The selective disadvantages in growth rate associated with plasmid carriage in the parent-like and ribitol dehydrogenase-overproducing strains was estimated. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 781-790 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; recombinant ; fed batch ; high cell density ; trypsin ; fermention ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fed-batch techniques were employed to obtain high cell density cultures (92-100 g DCW/L) of Escherichia coli strain X90 producing a recombinant serine protease, rat anionic trypsin, secreted to the periplasm. The specific growth rate was controlled to minimize growth-inhibiting acetate formation by utilizing an exponential feeding profile determined from mass balance equation. The volumetric yield of recombinant rat anionic trypsin was 56 mg/L, and the final cell density was 92 g DCW/L when the culture was induced in the late logarithmic phase. However, when the culture was induced in the early logarithmic phase, the volumetric yield was 13 mg/L and the final cell density was 14 g DCW/L. Thus, the induction timing is shown to have a significant effect on the final cell density as well as the overall volumetric yield of the recombinant protease. © 1993 Wiley & Sons, Inc.
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  • 61
    ISSN: 0006-3592
    Keywords: hybridoma ; kinetics ; curve fitting ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The use of partial cubic spline data interpolation for the calculation of volumetric metabolite exchange rates suggested the existence of three distinct metabolic phases during bioreactor culture of a hybridoma cell line. During phase 1, a rapid amino acid uptake rate and ammonia release rate were observed. The growth rate was low and glutamine synthetase activity fell. In phase 2, maximum growth rate and minimum glutamine assimilation and ammonium production rates were observed. Attempts to corroborate the apparent ammonia assimilation in this phase using 15NH4Cl resulted in low incorporation rates into alanine and glutamine. Maximum glutamine synthetase activity took place during this period. Maximum antibody production rate was observed during phase 3 during which peaks in glutamine assimilation, ammonia release, and glutamine synthetase activity were observed. The apparent existence of the three phases prompted us to carry out Northern blot analysis of glutamine synthetase RNA at appropriate times during the process. This revealed a pattern of appearance and dis-appearance of mRNA consistent with the three phases indicated by the fermentation parameters. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 42 (1993), S. 557-570 
    ISSN: 0006-3592
    Keywords: mathematical model of cell growth ; continuous culture ; protein excretion ; β-lactamase ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simple mathematical model is developed to help explain the complex population dynamics of an Escherichia coli host-plasmid expression/excretion system for β-lactamase within single- and two-stage reactors. The model successfully integrates the individual regulatory (tac promoter induction), genetic (runaway plasmid replication), and population dynamics (culture instability) aspects of the system. The model predicts, and experiment confirms, that high-level β-lactamase production and excretion cannot be easily maintained in single-stage reactors using the current plasmid construction. Stable target protein production and excretion is mathematically predicted, and experimentally confirmed, within two-stage reactors. The model is used to provide insight into engineering a more stable host-vector expression/excretion system for use in single-stage reactors. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 42 (1993), S. 625-634 
    ISSN: 0006-3592
    Keywords: polyunsaturated fatty acid (PUFA) ; microbial oil ; gamma linolenic acid (GLA) ; production ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The fatty acid production characteristics of fungi are described. These characteristics are the relationship between the oil content of the cell and the fatty acid content of the oil. For example, for polyunsaturated fatty acid (PUFA) production by Mucor hiemalis IPD 51, the oil content of the cell and the GLA content of the oil are coupled. For fungal production of some PUFA, synthesized after the rate-limiting step in the fatty acid anabolic chain, a maximum fatty acid production model was developed to link the fatty acid content of the oil and the oil content of the cell. Maximum volumetric productivity of gamma linolenic acid (GLA) by molds was found to occur at a specific GLA content of the oil. For example, for M. hiemalis IPD 51, a maximum volumetric of 4.7 mg GLA/L · h was produced at a GLA content of the oil of 8% to 10%. Similarly for Mucor circinelloides v. Tieghem IPD 155 a maximum volumetric productivity of 4.8 mg GLA/L · h was produced at a GLA content of the oil of 14% to 16%. These results imply that, when screening microorganisms for GLA or other fatty acid production, a number of medium compositions need to be evaluated to determine the tradeoff between oil content of the cell and fatty acid content of the oil. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 42 (1993), S. 1190-1198 
    ISSN: 0006-3592
    Keywords: fermentation ; bioprocess monitoring ; bioluminescence ; inner filter effect ; Escherichia coli ; cell concentration monitoring ; fiber optic ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Bioluminescence has recently become a popular research tool in several fields, including medicine, pharmacology, biochemistry, bioprocessing, and environmental engineering. Beginning with purely qualitative goals, scientists are now targeting more demanding applications where accurate, quantitative interpretation of bioluminescence is necessary. Using the recent advances in fiber-optic technology, bioluminescence is easily monitored in vivo and in real time. However, the convenience of this measurement is often concealing an unsuspected problem: the bioluminescence signal might be corrupted by a large error caused by the extinction of light by biological cells. Since bioluminescent cultures not only emit light but also absorb and scatter it, the measured signal is related in a complex, nonlinear, and cell-concentration-dependent manner to the “true” bioluminescence. This light extinction effect, known as the “inner filter effect,” is significant in high-density cultures. Adequate interpretation of the bioluminescence signal can be difficult without its correction. Here, we propose a real-time algorithm for elimination of the inner filter effect in a bioreactor. The algorithm yields the bioluminescence which would be measured if the glowing culture was completely transparent. This technique has been successfully applied to batch and continuous cultivation of recombinant bioluminescent Escherichia coli. © 1993 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 1092-1100 
    ISSN: 0006-3592
    Keywords: high cell density cultivation ; Escherichia coli ; XAD adsorbents ; dialysis reactor ; controlled substrate feed ; inhibitory products, removal of ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Reduction in nutrient loss during dialysis cultivation of Escherichia coli on a glycerol medium was investigated. A dialysis reactor with an inner fermentation and an outer dialysis chamber was used. Aerobic condition was maintained by limiting the glycerol feed rate to an optimum value which was estimated from the oxygen requirements for glycerol oxidation and oxygen transfer capacity of the reactor. High reduction in nutrient loss was achieved by using water as the dialyzing fluid. However, osmotic movement of water from the dialysis to the fermentation chamber was observed, and the final cell concentration was low. With a nutrient-split feeding strategy (feeding glycerol directly to the fermentation chamber and dialyzing with salt solution), glycerol loss was small, there was no osmotic flux of water to the fermentation chamber, and the cell concentration was high. Both glycerol and salt loss could be avoided, and a cell concentration of 170 g/L was obtained when the dialysis process was substituted by addition of XAD adsorbents to the dialysis chamber. Application of this nutrient-split feeding strategy to cell cultivation in a stirred tank reactor, coupled with dialysis in external dialyzer modules, resulted in low cell concentrations. © 1993 Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 41 (1993), S. 211-220 
    ISSN: 0006-3592
    Keywords: plasmid retention ; gene expression ; biofilm ; β-galactosidase ; segregational instability ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Differences in plasmid retention and expression are studied in both suspended and biofilm cultures of Escherichia coli DH5α(PMJR1750). An alternative mathematical model is proposed which allows the determination of plasmid loss probability in both suspended batch and continuously fed biofilm cultures. In our experiments, the average probability of plasmid loss of E. coli DH5α(pMJR1750) is 0.0022 in batch culture in the absence of antibiotic selection pressure and inducer. Under the induction of 0.17 MM IPTG, the maximum growth rate of plasmid-bearing cells in suspended batch culture dropped from 0.45 h-1 to 0.35 h-1 and the β-galactosidase concentration reached an experimental maximum of 0.32. pg/cell 4 hours after the initiation of induction. At both 0.34 and 0.51 mM IPTG, growth rates in batch cultures decreased to 0.16 h-1, about 36% of that without IPTG, and the β-galactosidase concentration reached an experimental maximum of 0.47 pg/cell 3 hours after induction.In biofilm cultures, both plasmid-bearing and plasmid-free cells in increase with time reaching a plateau after 96 hours n the absence of both the inducer and any antibiotic selection pressure. Average probability of plasmid loss for biofilm-bound E. coli DH5β(pMJR1750) population was 0.017 without antibiotic selection. Once the inducer IPTG was added, the concentration of plasmid-bearing cells in biofilm dropped dramatically while plasmid-free cell numbers maintained unaffected. The β-galactosidase concentration reached a maximum in all biofilm experiments 24 hours after induction; they were 0.08, 0.1, and 0.12 pg/cel under 0.17, 0.34, and 0.51 mM IPTG, respectively. © 1993 John Wiley & Sons, Inc.
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  • 67
    ISSN: 0006-3592
    Keywords: acetate production ; E. coli ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: An integrated metabolic model for the production of acetate by growing Escherichia coli on glucose under aerobic conditions is presented. The model is based on parameters which are easily determined by experiments. Forming the basis for this integrated metabolic model are the 12 principal precursor metabolites for biosynthetic pathways, the Embden-Meyerhof-Parnas pathway, the pentose phosphate cycle, the tricarboxylic acid cycle and the anapleurotic reactions, the Crabtree effect, the Pasteur effect, and the details of bacterial respiration. The result can be used to explain phenomena often observed in industrial fermentations, i.e., increased acetate production which follows from high glucose uptake rate, a low oxygen concentration, a high specific growth rate, or a combination of these conditions. © 1993 John Wiley & Sons, Inc.
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  • 68
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    Biotechnology and Bioengineering 41 (1993), S. 14-24 
    ISSN: 0006-3592
    Keywords: penicillin G amidase ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Penicillin G amidase (PGA) is a key enzyme for the industrial production of penicillin G derivatives used in therapeutics. Escherichia coli ATCC 11105 is the more commonly used strain for PGA production. To improve enzyme yield, we constructed various recombinant E. coli HB101 and ATCC 11105 strains. For each strain, PGA production was determined for various concentrations of glucose and phenylacetic and (PAA) in the medium. The E. coli strain, G271, was identified as the best performer (800 U NIPAB/L). This strain was obtained as follows: an E. coli ATCC 11105 mutant (E. coli G133) was first selected based on a low negative effect of glucose on PGA production. This mutant was then transformed with a pBR322 derivative containing the PGA gene. Various experiments were made to try to understand the reason for the high productivity of E. coli G271. The host strain, E. coli G133, was found to be mutated in one (or more) gene(s) whose product(s) act(s) in trans on the PGA gene expression. Its growth is not inhibited by high glucose concentration in the medium. Interestingly, whereas glucose still exerts some negative effect on the PGA production by E. coli G133, PGA production by its transformant (E. coli G271) is stimulated by glucose. The reason for this stimulation is discussed. Transformation of E. coli G133 with a pBR322 derivative containing the Hindlll fragment of the PGA gene, showed that the performance of E. coli G271 depends both upon the host strain properties and the plasmid structure. Study of the production by the less efficient E. coli HB101 derivatives brought some light on the mechanism of regulation of the PGA gene. © 1993 John Wiley & Sons, Inc.
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  • 69
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    Biotechnology and Bioengineering 41 (1993), S. 557-565 
    ISSN: 0006-3592
    Keywords: amino acid addition ; protein stability ; stress response ; Escherichia coli ; chloramphenicol-acetyl-transferase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this work, effective feeding schemes that would minimize stress responses to cloned-protein overexpression are investigated. The cloned-protein (chloramphenicolacetyl-transferase, CAT) contains a high aromatic amino acid content, most notably a high phenylalanine content. Experiments performed on Escherichia coli RR1 [pBR329] (constitutive promoter) and E. coli JM105 [pSH101] (inducible promoter) demonstrated that phenylalanine addition increases the rate of synthesis and yield of CAT. A previous study correlating inducer strength with CAT expression in E. coli JM105 [pSH101] indicated that the highest expression rate was accompanied by the highest apparent rate of protein degradation. In this work, the combined addition of isopropyl-β-D-thiogalactopyranoside (IPTG) and phenylalanine at intermediate levels resulted in substantial increase of CAT synthesis and partial reduction of protein degradation. Furthermore, transmission electron micrographs verified the absence of inclusion bodies, which, along with proteases, were suspected to reduce protein activity. The research demonstrates that significant enhancement in production and stability of heterologous proteins is possible by designing feeding strategies that incorporate knowledge of the interaction between primary cellular metabolism and foreign protein expression. © 1993 John Wiley & Sons, Inc.
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  • 70
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    World journal of microbiology and biotechnology 8 (1992), S. 71-72 
    ISSN: 1573-0972
    Keywords: Enzymes ; cassava ; fermentation ; ‘fufu’
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Amylase and pectin methyl esterase activities increased rapidly during the early period of the fermentation of cassava for ‘fufu’ production, attaining their peak activities after 12 and 24h, respectively. Cellulase activity was lower and approximately constant for most of the fermentation period.
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    World journal of microbiology and biotechnology 8 (1992), S. 175-182 
    ISSN: 1573-0972
    Keywords: Cassava ; cyanide ; fermentation ; fu-fu, gari ; lactic acid bacteria ; linamarin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Processes for the production ofgari, East Nigerianfu-fu and West Nigerianfu-fu are described. Changes in pH value, moisture content, microflora and sugar content of cassava duringgari andfu-fu preparation are reported. Mannitol accumulated during thegari fermentation but not in either of thefu-fu fermentations. During each stage ofgari andfu-fu production, lactic acid bacteria predominated. Homofermentative organisms occurred most frequently in the early stages of each process and heterofermenters in the latter ones. Of the 179 microorganisms that were isolated and characterized fromgari andfu-fu, 52% were able to hydrolyse linamarin and 14% starch.
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  • 72
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    World journal of microbiology and biotechnology 8 (1992), S. 284-286 
    ISSN: 1573-0972
    Keywords: Bacillus ; budu ; fermentation ; fish ; proteases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Eight different strains ofBacillus were isolated from fermented fish (Budu) and their proteolytic enzyme activities were determined after 18 h cultivation at room temperature (35° C). Four isolates possessed high protease activities. Optimum pH for these enzymes was between 7.0 and 8.0 and the optimal temperature was 55° C. The proteases retained 40% of their original activity after 20 min at 55° C but lost all activity at 65° C. Three of the four isolates were identified asBacillus subtilis, the fourth asBacillus licheniformis.
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  • 73
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    Biotechnology and Bioengineering 39 (1992), S. 607-613 
    ISSN: 0006-3592
    Keywords: lactic acid ; Lactobacillus delbreuckii ; extractive ; fermentation ; product inhibition ; packed-column and ion-exchange resin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Lactic acid fermentation is an end-product-inhibited reaction. The restriction imposed by lactic acid on its fermentation can be avoided by extractive fermentation techniques. Studies were performed by attaching an ion-exchange resin packed column with a 2-L fermentor for separation of lactic acid. The fermentation, in a conventional batch mode, resulted in a lactic acid yield of 0.828 g · g-1 and a lactic acid productivity of 0.313 g · L-1 · h-1. However, these could be further enhanced to 0.929 g · g-1 and 1.665 g · L-1 · h-1 by extractive fermentation techniques. The effect of temperature on extractive fermentation was remarkable and has been included in this work.
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  • 74
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    Biotechnology and Bioengineering 40 (1992), S. 875-888 
    ISSN: 0006-3592
    Keywords: cell concentration ; light scatter ; solid substrate ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel sensor was developed, based on light scatter, to estimate the cell concentration in the presence of suspended solids. The light scatter properties of cells in the presence of suspended solids were investigated. Two crucial observations were made: first, that the light scatter from cells is essentially a linear function of cell concentration and, second, that invariant regions are present in the light scatter spectrum of cell/solid substrate mixtures. Invariant regions are wavelength intervals of the light scatter spectrum in which the light scatter reading is independent of solid substrate concentration and only a function of cell concentration. The occurrence of invariant regions is the key behavior which allowed the quantification of cell concentration in the presence of suspended solids.An algorithm was developed for the estimation, from light scatter data, of cell concentration in the presence of solid substrate. The light scatter approach was validated by comparing cell concentrations estimated by this technique to those obtained from DNA and carbon dioxide evolution rate measurements during a series of fermentations. The model system used was Bacillus subtilis var sakainensis ATCC 21394 growing on fishmeal as the sole nitrogen source.A model was developed based on the interactions of scatter and absorbance. This model reflects the hypothesis that invariant regions are caused by changes in the absorbance of the solid substrate as a function of wavelength. © 1992 John Wiley & Sons, Inc.
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  • 75
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    Biotechnology and Bioengineering 39 (1992), S. 663-671 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; acetic acid ; methionine ; yeast extract ; continuous fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Acetic acid formation in Escherichia coli fermentation has been studied in continuous cultures. Experimental results suggest that the limited capacity of the oxidative metabolism (perhaps the limited capacity of TCA cycle) may be responsible for acetic acid formation. At low growth rates, both anabolic and catabolic requirements may be satisfied by the oxidative metabolism. However, at high growth rates these two demands may exceed the capacity of the oxidative metabolism alone. It is proposed that under these circumstances, E. coli reorganizes the oxidative metabolism to first meet the anabolic requisition and then supply the necessary amount of energy using both the remaining capacity of the oxidative metabolism and acetic acid formation metabolism. Escherichia coli selects acetic acid synthesis as the aerobic energy source because it generates the second largest amount of ATP and NADH2. According to our proposition, acetic acid formation could be reduced by decreasing the anabolic requirement, i.e., reducing glucose uptake, or by increasing the capacity of the oxidative metabolism. These two approaches were experimentally confirmed by observing reduced acetic acid formation by reducing the glucose uptake with a yeast extract addition and enhancing the capacity of oxidative metabolism with a methionine addition.
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    Biotechnology and Bioengineering 39 (1992), S. 732-740 
    ISSN: 0006-3592
    Keywords: cell disruption ; chemical permeabilization ; Escherichia coli ; fermentation ; protein recovery ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Overall protein release greater than 75% in less than 1 h can be attained by exposing exponentially growing Escherichia coli cells to 0.4 M guanidine plus 0.5% Triton X-100 at 37°C in medium. Cell growth stops immediately upon addition of the chemicals, but the cells are not lysed. Guanidine concentrations lower than 0.2 M, in conjunction with 0.5% Triton X-100, do not release significant intracellular protein, nor do they inhibit cell growth. Under these conditions, the cells undergo an adaptation that confers resistance to protein release by further treatment with guanidine and Triton X-100. Cells treated with 0.2 M guanidine plus 0.5% Triton X-100 display intermediate behavior. Protein release is approximately 35%, and growth is temporarily interrupted by an extended lag phase. Subsequent resumption of cell growth results in resistant cells and no additional protein release. This resistance is shown to be reversible and is most likely due to physiological adaptation rather than genetic mutation.
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    Biotechnology and Bioengineering 39 (1992), S. 775-780 
    ISSN: 0006-3592
    Keywords: formate ; Escherichia coli ; formate hydrogenlyase ; cell immobilization ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Formate hydrogenlyase (FHL) activity was induced in a strain of Escherichia coli S13 during anaerobic growth in yeast extract-tryptone medium containing 100 mM formate. The cells obtained at the optimum growth phase were immobilized in 2.5% (w/v) agar gel when 50-60% of the whole cell FHL activity was retained. The immobilized FHL system had good storage stability and recycling efficiency. In the lysis of formate, an increase of formate concentration to 1.18M increased QH2 (initial) value of the immobilized cell, and subsequently cells, hydrogen evolution, in general, ceased after 6 to 8 of incubation, resulting in incomplete lysis of formate. Presence of small amount of glucose (28 mM) was more or less quantitatively lysed with concomitant disappearence of glucose from the medium. Synthesis of formate from hydrogen and bicarbonate solution by the immobilized cells was also characterized. Presence of glucose (10 mM) in 50 mM bicarbonate solution stimulated formate synthesis by immobilized cells. The pH optimum range, Km, and specific activity of the immobilized cells for the lysis of formate were 6.8-7.2 0.4M, and 66 mL/g cell-h, respectively. The cells could fix hydrogen to the extent of 24.4% (w/w) of its own wet cell mass in a 72-h reaction cycle. Potentiality of the immobilized FHL system for biotechnological exploitation was discussed.
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  • 78
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    Biotechnology and Bioengineering 40 (1992), S. 271-279 
    ISSN: 0006-3592
    Keywords: carbon starvation ; Escherichia coli ; growth control ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The use of glucose starvation to uncouple the production of recombinant β-galactosidase from cell growth in Escherichia coli was investigated. A lacZ operon fusion to the carbon starvation-inducible cst-1 locus was used to control β-galactosidase synthesis. β-Galactosidase induction was observed only under aerobic starvation conditions, and its expression continued for 6 h following the onset of glucose starvation. The cessation of β-galactosidase expression closely correlated with the exhaustion of acetate, an overflow metabolite of glucose, from the culture medium. Our results suggest the primary role of acetate in cst-1-controlled protein expression is that of an energy source. Using this information, we metered acetate to a glucose-starved culture and produced a metabolically sluggish state, where growth was limited to a low linear rate and production of recombiant β-galactosidase occurred continuously throughout the experiment. The cst-1 controlled β-galactosidase synthesis was also induced at low dilution rates in a glucose-limited chemostat, suggesting possible applications to high-density cell systems such as glucose-limited recycle reactors. This work demonstrates that by using an appropriate promoter system and nutrient limitation, growth can be restrained while recombinant protein production is induced and maintained.
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    Biotechnology and Bioengineering 39 (1992), S. 225-232 
    ISSN: 0006-3592
    Keywords: electroconductive heating ; electrical pretreatment ; thermal death kinetics ; zygo Saccharomyces bailii ; Escherichia coli ; microorganisms ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Suspensions of yeast cell (zygo Saccharomyces bailii) in a phosphate buffer solution were subjected to conventional (hot water) and ohmic (electric current) heating under identical temperature histories. Experiments were also conducted with cells of Escherichia coli to compare the lethal effect of combination of sublethal electrical preteatment and conventional heating with conventional heating. The kinetic parameters (D,Z,K and Ea) were determined for both organisms during different treatments. There was no significant difference in the death rate of yeast cells during conventional and ohmic heating at the voltage range used in this study. Results of electrical pretreatment and conventional heating on E. coli indicated differences under certain conditions when compared with pure conventional heating. Thus it is concluded that microbial death during ohmic heating was due primarily to thermal effects with no significant effect of electric current per se. Sublethal electrical pretreatment appears to offer potential for increased bacterial inactivation in certain cases.
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    Biotechnology and Bioengineering 39 (1992), S. 293-304 
    ISSN: 0006-3592
    Keywords: control ; fermentation ; baker's yeast ; high cell density ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A methodology for the design and evalution of bioprocess control strategies is presented. The strategies manage nutrient supply with demand and vary with the metabolic condition and phase of fermentation operation. Six carbon source addition strategies are based on different combinations of available measurements; they are described and evaluated under different operating conditions for yeast cultivation. It is concluded that a single control strategy is not the most appropriate under all possible operating conditions. An oxygen uptake rate-based control strategy performs better with a mean respiratory quotient (RQ) value less than 1.1 during an oxygen limitation than an ethanol control strategy which had a mean RQ of 14. The designed strategies and an approach of applying the strategy that best matches fermentation conditions consistently enables achievement of high cell densities 78.7 g DCW/L and yields 0.50 g DCW/g glucose as the mean values for three fermentations.
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    Biotechnology and Bioengineering 39 (1992), S. 365-368 
    ISSN: 0006-3592
    Keywords: fermentation ; adsorption ; lactic acid ; fluidized bed ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A bioreactor configuration is proposed for simultaneous fermentation and separation of the desired product. The bioreactor consists of a columnar fluidized bed of immobilized microorganisms. Denser adsorbent particles are added to this column. These adsorbent particles fall through the bed, absorb the product, and are removed from the base of the columnar reactor. The system hydrodynamics and the separability of the two types of particles were confirmed for low-density gel beads. The addition of the adsorbent, activated carbon, to a fermentation of Lactobacillus delbreuckii absorbed lactic acid. The addition of adsorbent enhanced the fermentation and controlled the pH.
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  • 82
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    Biotechnology and Bioengineering 39 (1992), S. 398-407 
    ISSN: 0006-3592
    Keywords: recombinant bacterium ; plasmid stability ; filtering ; smoothing ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A numerical method to process experimental data concerning plasmid stability of a recombinant bacteria during continuous cultures with nonselective media is proposed here. This method differs from previous ones in that it uses the derivatve form of the state equation of the Imanaka-Aiba model for recombinant cultures. The methodology proposed here allows one to estimate values for the two model parameters without forcing them to be constant. Until now, this could not be done using classical analytical techniques because these parameters have been considered invariable because of the integration used in the evaluation of the model. These parameters are (1) the difference in the specific growth rates between plasmid-carrying cells and plasmid-free cells (δμ), and (2) the probability of plasmid loss by plasmid-containing cells (ρr μ+). The derivative technique used here is completed by mathematical treatments involving data filtering and smoothing. The values of the two parameters are in agreement with those already publised. The current technique does not impose preconditions and permit us to further study related phenomena.
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    Biotechnology and Bioengineering 40 (1992), S. 137-146 
    ISSN: 0006-3592
    Keywords: anomeric specificity ; mechanism of glucose uptake ; Lactococcus cremoris ; Escherichia coli ; Saccharomyces cerevisiae ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The mechanism and kinetics of the glucose uptake systems of three representative microorganisms are studied during cultivation in a chemostat. The three microorganisms are Lactococcus cremoris, Escherichia coli, and Saccharomyces cervisiae. Two models describing respectively competitive and independent uptake of the two glucose anomers are tested on experimental data where α- and β-glucose are determined by flow injection analysis after pulse addition of the pure anomers to a chemostat. The very accurate experimental results are used to give a convincingly clear model discrimination for all three microorganisms. The uptake of glucose by S. cervisiae occurs by a competitive mechanism with preference for α-glucose (Kα = 32 mg/L and Kβ = 48 mg/L). Surprisingly, the glucose uptake by the two bacteria is shown to be mediated by anomer specific transport systems with no competitive inhibition from the other glucose anomer. This novel finding has not been described in the literature on the phosphotransferase system. In L. cremoris the relative uptake rates of the glucose anomers match the equilibrium composition exactly (36% α-glucose). In E. coli the relative uptake rate of α-glucose at glucose unlimited growth is 26%, which means preference for β-glucose. However, the saturation constants of the two sites in E. coli are Kα = 2 mg/L and Kα = 15 mg/L, and a preference for α-glucose is exhibited at very low glucose concentrations. The results are of considerable improtance in relation to enzyme based on-line measurements during fermentations as well as to the modeling of glucose limited growth and product formation.
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    Biotechnology and Bioengineering 39 (1992), S. 815-827 
    ISSN: 0006-3592
    Keywords: Penicillium roquefortii ; sporulation ; solid substrate ; fermentation ; buckwheat ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: When Pencillium roquefortii is grown on two kinds of buckwheat, exhibiting a low [0.85 g water/g dry matter (DM), buckwheat A] and a high [1.5 g water/g initial dry matter (IDM), buckwheat B] water content, a marked difference in the mode of development of the fungus is observed. Material balances for buckwheat A show that growth does not stop because of nutrient exhaustion. Analysis of water balance shows that active growth proceeds with a permanent limitation by the turgor potential which disappears when the water activity of the substrate is close to 0.96, thus arresting growth. This limitation causes intensive water excretion from the system due to the lowering of the water activity of the substrate. The water content of the mycelium thus decreases from 79% at the beginning of the cultivation to 74% when the growth stops. This is linked to a substantial oxidative metabolism and a high sporulation efficiency, close to 0.85. The spores obtained have a low dry weight and a reduced nitrogen content. In the case of buckwheat B, the active growth is shown to stop because of available mineral nitrogen depletion. No significant decrease in the water activity of the substrate is found during the protein synthesis, and the turgor potential remains high at the end of this period. Culture proceeds with new wall synthesis; the sporulation efficiency remains high and the spores obtained exhibit a high dry weight and a high nitrogen content. The bioenergetic balances show that the P/O ratio varies with the kind of substrate used; its value is close to 1.56 for the low water medium and to 2.21 for the high one. The ATP yield Z is always close to 1, and fungal development occurs with limitations of both anabolism and catabolism on buckwheat B and only of anabolism and catabolism on buckwheat B and only of anabolism on buckwheat A.
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    Biotechnology and Bioengineering 39 (1992), S. 898-902 
    ISSN: 0006-3592
    Keywords: diffusion coefficient ; fermentation ; Aspergillus niger ; solid state fermentor ; ion-exchange resin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simple experimental diffusion controlled fermentor (DCF), coupled with the use of a mathematical model based on mass balance, is proposed to measure the variation of the gas (CO2 and O2) diffusion coefficients in solid state fermentation. The DCF was packed with an ion-exchange resin impregnated with a nutritive medium and inoculated with Aspergillus niger. The growth conditions in the DCF were very similar to those found in equipment operated with convective oxygen supply. The diffusion coefficient was shown to be very dependent on the biomass concentration within the solid state fermentor, and attained values of less than 5% of the molecular diffusion in air when the biomass in the fermentor reached 27 mg dry/g dry support.
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    Biotechnology and Bioengineering 40 (1992), S. 334-336 
    ISSN: 0006-3592
    Keywords: sampling, automatic containment ; Escherichia coli ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A containment sampling system for shake flasks and fermentors has been developed from a blood collection system used in hospitals. The core of the system is a collection vial with a vacuum inside. When a needle connected to the fermentation fluid penetrates a rubber seal on the vial, a sample is withdrawn. The system has been developed in two versions, a manual method for shake flasks, and an automated version for fermentors including cool storage of samples. The sampling system offers the same safety for fermentation containment as the original system offers safety for patients and hospital staff. © 1992 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 40 (1992), S. 634-637 
    ISSN: 0006-3592
    Keywords: mass spectrometer ; fermentation ; oxygen uptake rate ; noise ; bias ; filter ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The calculation of many derived fermentation variables such as the respiratory quotient (RQ) and mass transfer coefficient (KLa) requires the differences between the molar percentages of oxygen and carbon dioxide in the fermentor inlet and exit gas, called the %OUR and %CER. Noise and bias in %CER data is of order that in the exit gas carbon dioxide analysis. However, the relative amount of noise in the %OUR is one to two orders of magnitude greater than the noise in the raw oxygen analyses because the %OUR is calculated as a small difference between two large quantities. The noise in the %OUR is white with a Gaussian amplitude probability distribution of absolute standard deviation 0.0145. A chi-square filter of the %OUR data is shown to considerably improve the quality of the calculated RQ and KLa for a fermentation of Streptomyces clavuligerus. © 1992 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 40 (1992), S. 1197-1202 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; plasmid retention ; amino acid supplementation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of amino acid supplementation on plasmid stability in Escherichia coli B/r was tested experimentally. Comparisons of experimental results to computer-predicted values were made using a detailed, structured single-cell model. The plasmid, pDW17 (a pBR322 derivative with a mutated tac promoter controlling the β-lactamase gene), was used. In chemostat cultures, the amino acid supplemented cultures were always less stable than those grown in minimal medium. This effect was not a growth rate effect, as increasing growth rate imsproves stability for both cultures in minimal medium and in amino acid supplemented medium. The computer model also predicted a decrease in stability due to amino acid supplementation. The model also predicts that amino acid supplementation, combined with moderately strong plasmid-encoded protein expresion, results in a depletion of low-molecular-weight organics compared with plasmid-free cells. In minimal medium the same level of plasmid-encoded protein synthesis results in a strong reduction in amino acid pools compared with plasmid-free cells. With amino acid supplementation the growth differential between plasmid-bearing and plasmid-free cells may be due to an “energy limitation,” while in minimal medium the size of the growth rate differential may be due to a “building block” limitation. © 1992 John Wiley & Sons, Inc.
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  • 89
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    Biotechnology and Bioengineering 40 (1992), S. 1282-1285 
    ISSN: 0006-3592
    Keywords: Candida shehatae ; ethanol ; D-xylose ; viability ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Ethanol was added at concentrations of 25 and 50 g/L to active cultures of Canida shehatae under oxygen-limited (fermentative) conditions. Added ethanol completely inhibited grwoth and fermentation of D-xylose by C. shehatae. Cultures with added ethanol rapidly declined in cell viability as measured by plate counts and methylene blue staining. The rate of decline in cell viability was dependent on the amount of added ethanol. Over the course of the fermentation, cell viability, as measured by plate counts, was significantly lower in all experiments (with or without ethanol addition) compared with the viability measurements by methylene blue staining. Thus, data from the plate counts provided a more sensitive measure of the toxic effects of added ethanol and long-term anaerobiosis on C. shehatae growth/fermentation. Mean cell volume and total cell volume declined in fermentations with added ethanol. © 1992 John Wiley & Sons, Inc.
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  • 90
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    Biotechnology and Bioengineering 40 (1992), S. 1309-1318 
    ISSN: 0006-3592
    Keywords: Saccharomyces cerevisiae ; fermentation ; cell wall ; surface electrochemistry ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The cell-wall properties of three strains of the yeast Sacharomyces cerevisiae have been experimentally studied at various times during fermentation. The cell walls have been characterized by electrophoretic mobility measurements, from which zeta potentials may be calculated. They have also been characterized by computerized pH titration, which gives direct information on the number and nature of groups in the yeast cell wall. The data have been quantitatively analyzed in three ways. First, a simplified analysis of the electrokinetic data of a type used by previous workers has been applied. Second, such a simplified analysis of the electrokinetic data has been developed more rigorously by means of a two-dimensional site-dissociation model of the outer cell wall-solution interface. Third, a description of the yeast cell-wall electrochemical properties in terms of a three-dimensional gel model incorporating site dissociation has been developed. The advantages and disadvantages of the three analyses are discussed. Only the three-dimensional gel model can account simultaneously for both the electrokinetic and pH surface titration data. It provides new insights into the changes that occur to the yeast cell wall during fermentation. © 1992 John Wiley & Sons, Inc.
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  • 91
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    Biotechnology and Bioengineering 39 (1992), S. 1151-1160 
    ISSN: 0006-3592
    Keywords: fluorescence ; monitoring ; methane ; fermentation ; NAD(P)H ; F420 ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: On-line in situ fluorescence measurements of the methanogenic fermentation were conducted with reactors receiving either glucose or a mixture of volatile fatty acids as the substrate. The reactors were perturbed from steady-state conditions in order to assess the response of fluorescencemonitoring probes. Two fluorescence-monitoring probes were evaluated over a period of 8 months; they performed in a consistent manner, and their response was not significantly affected by the changes in pH and redox potential encountered during routine reactor operation. A commercially available probe, designed to measure NAD(P)H, demonstrated particular promise for detecting imbalance caused by the entry of air, inhibitor addition and was capable of distinguishing between different substrates. This fluorescence-monitoring probe detected imbalance more rapidly than other on-line measurements such as pH, Eh, or gas production, or off-line measurements such as volatile fatty acid concentration or gas composition. An experimental fluorescence-monitoring probe, designed to measure coenzyme F420, also showed some promise in this regard. The response of the fluorescence-monitoring probes also revealed details of the metabolic routes in the reactors and the probes represent a useful research tool. For example, a failure to observe the characteristic response of the NAD(P)H-monitoring probe to formate addition during the metabolism of acetate, propionate, or glucose strongly suggests that any formate liberated during their catabolism is degraded via a different route to exogenously added formate.
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  • 92
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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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  • 93
    ISSN: 0006-3592
    Keywords: homogenization, high-pressure ; cell disruption ; inclusion bodies ; size distribution ; centrifuge, analytical ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The high-pressure homogenization of Escherichia coli, strain JM101, containing inclusion bodies of recombinant porcine somatotropin was investigated. A novel technique employing an analytical disc centrifuge was used to monitor the disruption. This a direct technique which measures cell disintegration rather than soluble protein release. The technique is particularly suited to measurements where the disruption approaches 100%. The disk centrifuge provides a size distribution of the homogenate, and furnishes evidence for the preferential disruption of larger cells. For E. coli containing inclusion bodies, and increase in the cell feed concentration from 145 g/L (wet weight) to 330 g/L resulted is poorer homogenization. Poorer disruption was also obtained by lowering the feed temperature from 20°C to 5°C. Only slight variations in performance were obtained by increasing the feed pH from 7.5 to 9.0 or by storing the feed at 4°C for 24 h prior to disruption. Comparison with uninduced E. coli strain JM101, showed that the disruption obtained is higher for bacteria containing a recombinant inclusion body.
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  • 94
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    Biotechnology and Bioengineering 37 (1991), S. 985-988 
    ISSN: 0006-3592
    Keywords: Saccharomycopsis lipolytica ; rate equations ; fermentation ; citric acid ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 95
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    Biotechnology and Bioengineering 37 (1991), S. 1087-1094 
    ISSN: 0006-3592
    Keywords: fermentation ; optimization ; enzymatic reactions ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Two general models for batch simultaneous enzymatic and microbial reaction (SEMR) processes are presented, the second derived from and simpler than the first and accounting for enzyme denaturation. Using the second model and parameter values from the literature, simulation was used to examine a range of enzyme addition rate strategies (in which the rate was a linear function of time) for a relatively fast ethanol fermentation and for a longer duration citric acid fermentation, both using cellulose as the substrate. For the ethanol process it is optimal (for a specific objective function which accounts for product value and enzyme cost) to add all the enzyme at the beginning of the process. But for the citric acid process a linearly decreasing enzyme addition rate, coupled with the addition of a small fraction of the enzyme at time zero, is better than pure batch operation or operation with the best constant enzyme feed rate.
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  • 96
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    Biotechnology and Bioengineering 37 (1991), S. 1101-1107 
    ISSN: 0006-3592
    Keywords: simvastatin ; microbial ; hydroxylation ; fermentation ; biotransformation ; scale-up ; dissolved oxygen ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article describes a process for microbial hydroxylation of simvastatin by a Nocardia sp. Simvastatin (Zocor) belongs to the family of HMGCoA reductase inhibitors used as cholesterol-lowering drugs. Studies at 14 L scale showed that high substrate (simvastatin) concentrations inhibited product formation; consequently, continuous slow feeding of the substrate was introduced to maintain low residual simvastatin concentrations. Dissolved oxygen levels above 50% air saturation were desirable for the biotransformation. The process was scaled up to 19,000-L fermentors using an on-line filter sterilization system for substrate feeding. The feed rate was regulated by off-line high-pressure liquid chromatography (HPLC) assays to keep the substrate concentration below 20 mg/L. Intermittent addition of nutrients helped to boost the bioconversion rate to give final titers of 400 mg/L 6-β-hydroxymethyl simvastatin. Enrichment of the nutrient medium led to bioconversion titers of 800 mg/L 6-β-hydroxymethyl simvastatin. Bioconversion efficiencies (desired product/substrate) of 22-25% with a ratio of desired product/side products of 0.7 were obtained by this process.
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  • 97
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    Biotechnology and Bioengineering 38 (1991), S. 11-23 
    ISSN: 0006-3592
    Keywords: lactic acid fermentation ; fermentation ; microbial fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A two-compartment model for the lactic acid fermentation with Streptococcus cremoris is experimentally verified. The seven parameters of the model are determined using steady-state chemostat data at varying values of dilution rate, D, but with a constant feed concentration, sf, of a single carbohydrate source (glucose, lactose, or galactose), and a constant feed concentration of sNf of the N source. Steady-state measurements of the RNA content at different exit concentrations, s, of the carbohydrate are included to calculate kinetic parameters that determine the cell composition for varying operating conditions. The model is tested using data from a large set of steady-state and non-steady-state experiments: batch fermentations and step and pulse experiments in a chemostat. Both qualitatively and quantitatively the major features of the model are confirmed: the external substrates enter into intracellular high-energy building blocks, and lactic acid is formed as a by-product of these reactions. Cell growth depends on the fraction of active components (XA) of the cell and is not accompanied by lactic acid production. Possible model modifications are discussed, primarily to obtain a better description of lactic acid fermentation at nongrowth conditions.
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  • 98
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    Biotechnology and Bioengineering 37 (1991), S. 703-707 
    ISSN: 0006-3592
    Keywords: Dextransucrase ; Leuconostoc mesenteroides ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: High yields of the enzyme dextransucrase have been produced repeatedly by fed-batch fermentation techniques. Activities in excess of 21.9 U/cm3 have been obtained by culturing Leuconostoc mesenteroides NRRL B-512(F) under nonaerated fed-batch fermentation conditions. Aerobic fermentations carried out under identical conditions have consistently produced enzyme of less than 17 U/cm3, but with no difference in the final cell concentration in the broth. Different types of yeast extract have been found to have significant effect on the final cell concentration and more especially on the enzyme activity with enzyme yields varying by as much as 50% when different types of yeast extracts were used.
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  • 99
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    Biotechnology and Bioengineering 37 (1991), S. 869-875 
    ISSN: 0006-3592
    Keywords: scu-PA ; pro-urokinase ; yeast ; respiratory quotient ; fermentation ; Saccharomyces cerevisiae ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Secretion of a nonglycosylated form of human pro-urokinase, also known as single-chain urinary plasminogen activator (scu-PA), from Saccharomyces cerevisiae is described. A “supersecreting” yeast strain harboring multiple copies of integrated plasmids was grown batchwise and at constant respiratory quotient (RQ) in 20-L fermenters. Because the promoters used to drive expression of the pro-urokinase genes are not tightly regulated, secretion into the culture supernatant was growth associated. Although the final cell density achieved in the perturbed-batch fermentation (45 g dry wt/L) was less than that observed in the RQ-controlled culture (77 g dry wt/L), the scu-PA titer in the perturbed-batch fermentation (1863 IU/mL) was nearly twice that attained at constant RQ (1108 IU/mL). The effects on cell growth and scu-PA titer of other process variables (pH, temperature, phosphate concentration, and medium composition) are also discussed.
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  • 100
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    Biotechnology and Bioengineering 38 (1991), S. 314-318 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; maltoporin ; harvesting bacteria ; bacterial surfaces ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Addition of starch to suspensions of Escherichia coli K-12 resulted in the formation of bacterial flocs. The flocculation was dependent on the high expression of a receptor for starch (maltoporin) on the surface of the bacterium. Factors influencing floc formation were investigated and optimal conditions for flocculation based on cell density, starch concentration, time, and pH established. As quantitated by a sedimentation assay, over 80% of bacteria in a culture could be removed by settling without centrifugation in 3 h under optimal conditions. Floc formation was evident with bacteria containing wild-type maltoporin but was faster and occurred to a greater extent with strains expressing a high-affinity allele (lamB1400) of the starch receptor. Bacteria could be harvested by floc formation directly in growth medium under defined conditions of maltoporin expression and medium composition. These results demonstrate the effectiveness of starch-dependent aggregation in the harvesting of cells, using an inexpensive, biologically acceptable agent to induce flocculation.
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