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  • Articles  (1,231)
  • Industrial Chemistry and Chemical Engineering  (1,133)
  • Escherichia coli  (98)
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  • 1
    ISSN: 0006-3592
    Keywords: Escherichia coli ; Chloramphenicol Acetyltransferase (CAT) ; Culture Redox Potential (CRP) ; Dithiothreitol (DTT) ; reducing agents ; molecular chaperones ; proteases ; heat shock ; stress response ; protein folding ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The independent control of culture redox potential (CRP) by the regulated addition of a reducing agent, dithiothreitol (DTT) was demonstrated in aerated recombinant Escherichia coli fermentations. Moderate levels of DTT addition resulted in minimal changes to specific oxygen uptake, growth rate, and dissolved oxygen. Excessive levels of DTT addition were toxic to the cells resulting in cessation of growth. Chloramphenicol acetyltransferase (CAT) activity (nmoles/μg total protein min.) decreased in batch fermentation experiments with respect to increasing levels of DTT addition. To further investigate the mechanisms affecting CAT activity, experiments were performed to assay heat shock protein expression and specific CAT activity (nmoles/μg CAT min.). Expression of such molecular chaperones as GroEL and DnaK were found to increase after addition of DTT. Additionally, sigma factor 32 (σ32) and several proteases were seen to increase dramatically during addition of DTT. Specific CAT activity (nmoles/μg CAT min.) varied greatly as DTT was added, however, a minimum in activity was found at the highest level of DTT addition in E. coli strains RR1 [pBR329] and JM105 [pROEX-CAT]. In conjunction, cellular stress was found to reach a maximum at the same levels of DTT. Although DTT addition has the potential for directly affecting intracellular protein folding, the effects felt from the increased stress within the cell are likely the dominant effector. That the effects of DTT were measured within the cytoplasm of the cell suggests that the periplasmic redox potential was also altered. The changes in specific CAT activity, molecular chaperones, and other heat shock proteins, in the presence of minimal growth rate and oxygen uptake alterations, suggest that the ex vivo control of redox potential provides a new process for affecting the yield and conformation of heterologous proteins in aerated E. coli fermentations. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59: 248-259, 1998.
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  • 2
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    Biotechnology and Bioengineering 51 (1996), S. 434-438 
    ISSN: 0006-3592
    Keywords: polyphosphate ; Escherichia coli ; phosphate starvation ; gene expression ; heterologous ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of intracellular polyphosphate on the phosphate-starvation response in Escherichia coli was studied by genetically manipulating the intracellular polyphosphate levels and by performing phosphate shifts on the genetically engineered strains. Strains that produced large quantities of polyphosphate and were able to degrade it induced the phosphate-starvation response to a lesser extent than wild-type strains, whereas strains that were unable to degrade a large intracellular polyphosphate pool induced the phosphate-starvation response to a greater extent than wild-type strains. These results have important implications for expression of heterologous genes under control of the phoA promoter. © 1996 John Wiley & Sons, Inc.
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  • 3
    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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  • 4
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    New York, NY [u.a.] : Wiley-Blackwell
    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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  • 5
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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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  • 6
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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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  • 7
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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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  • 8
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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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  • 9
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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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  • 10
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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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  • 11
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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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  • 12
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    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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  • 13
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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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  • 14
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    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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  • 15
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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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  • 16
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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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  • 17
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    Biotechnology and Bioengineering 37 (1991), S. 661-672 
    ISSN: 0006-3592
    Keywords: bacterial chemotaxis ; Escherichia coli ; random motility ; diffusion chamber assay ; mathematical model ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A quantitative description of bacterial chemotaxis is necessary for making predictions about the migratory behavior of bacterial populations in applications such as biofilm development, release of genetically engineered bacteria into the environment, and in situ bioremediation technologies. The bacterial chemotactic response is characterized by a mathematical model which relates individual cell properties such as swimming speed and tumbling frequency to population parameters, specifically the random motility coefficient and the chemotactic sensitivity coefficient. Our model includes a nonlinear dependence of the chemotactic velocity on the attractant gradient as well as a dependence of the random motility coefficient on the temporal and spatial attractant gradients, both of which previous analyses have neglected. As we will show, these aspects are critical for interpreting the results from experiments like those performed in the stopped-flow diffusion chamber (SFDC) because the initial temporal and spatial gradients are very steep. Our analysis demonstrates that values for the random motility coefficient and chemotactic sensitivity coefficient can be obtained from experimental plots of net cell redistribution from initial conditions versus the square root of time. Values for these parameters are determined from experimental measurements of bacterial population distributions in the SFDC as described in the companion article. Using parameter values determined from independent experiments, μ = 1.1 ± 0.4 ± 10-5 cm2/s and χ0 = 8 ± 3 ± 10-5 cm2/s, excellent agreement is found between theoretically predicted bacterial density profiles and actual experimental profiles for Escherichia coli K12 responding to fucose over two orders of magnitude in initial attractant concentration. Thus, our model captures the concentration dependence of this behavioral response satisfactorily in terms of cell population parameters which are derived from individual cell properties and will therefore be useful for making predictions about the migratory behavior of bacterial populations in the environment.
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  • 18
    ISSN: 0006-3592
    Keywords: Escherichia coli ; growth factor ; epidermal growth factor ; fed batch culture ; human epidermal growth factor ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fed-batch cultures of recombinant E. coli HB101 harboring expression plasmid pTRLBT1 or pTREBT1, with acetate concentration monitoring, are investigated to obtain high cell density and large amounts of human epidermal growth factor (hEGF). The expression plasmid pTRlBT1 contains a synthetic hEGF gene attached downstream of the N-terminal fragment of the trp L gene preceded by the trp promoter. The expression plasmid pTREBT1 contains the same coding sequence attached downstream of the N-terminal fragment of the trp E gene preceded by the trp promoter, trp L gene, and attenuator region. E. coli harboring pTREBT1 produces 0.56 mg/L hEGE and immediately degrades it. On the other hand E. coli harboring pTRLBT1 produces 6.8 mg/L hEGF and does not decompose it. Prominent inclusion bodies are observed in E. coli cells harboring pTRLBT1 using an election microscope. To Cultivate E. coli harboring pTRLBT1, a fed-batch culture system, divided into a cell growth step and an hEGF production step, is carried out. The cells grow smoothly without acetate-induced inhibition. Cell concentration and hEGF quantity reach the high values of 21 g/L and 60 mg/L, respectively.
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  • 19
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    Biotechnology and Bioengineering 38 (1991), S. 831-837 
    ISSN: 0006-3592
    Keywords: fermentation ; Escherichia coli ; recombinant fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of dilution rates on the performance of a two-stage fermentation system for a recombinant Escherichia coli culture were studied. Dilution rate determines the apparent or averaged specific growth rate of a heterogeneous population of cells in the recombinant culture. The specific growht rate affects the genetic parameters involved in product formation in the second stage, such as plasmid stability, plasmid content, and specific gene expression rate. Kinetic models and correlations were developed for these parameters based on experimental data. Simulations of plasmid stability in the first stage showed that for longer fermentation periods, plasmid stability is better at higher dilution rates. However, the plasmid content is lower at these dilution rates. The optimal apparent specific growth rate for maximum productivity in the second stage was determined using two methods: (1) direct search for a constant specific growth rate, and (2) dynamic optimization using the maximum principle for a time-dependent specific growth rate profile. The results of the calculations showed that the optimum constant apparent specific growth rate for maximum over-all productivity is 0.40 h-1. This coincides with the optimal specific growht rate for maximum plasmid content in the expressed stage. A 3.5% increase in overall productivity can be obtained by using a linear time dependent apparent specific growth rate control, μ2(t) = 0.0007t, in the course of the fermentation time.
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  • 20
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    Biotechnology and Bioengineering 38 (1991), S. 891-906 
    ISSN: 0006-3592
    Keywords: ribosome vector ; cloned-gene expression ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An expression system utilizing specialized ribosomes has been constructed with β-galactosidase as the product. Ribosomes specific for lacZ mRNA are generated due to a mutation within the anti-Shine-Dalgarno region of a plasmidborne 16S rRNA gene that is complementary to a mutation within the ribosome-binding site of lacZ. Hence, a subpopulation of ribsomes specific for translation of the cloned gene mRNA is produced. Transcription of the lacZ gene is regulated by the tac promoter, while transcription of the mutated rrnB locus is controlled by the λPL promoter. Batch experiments indicate that full induction of both operons (2 mM IPTG, 42°C) leads to maximal β-galactosidase activity per cell at levels 35% higher than that obtained using a wild-type ribosome expression system. Using a novel, site-directed mutagenesis technique, construction of the specialized ribosome vector is outlined, and the results of lacZ expression are presented as transcription of both the cloned-gene and the specialized-ribosome locus are induced.
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  • 21
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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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  • 22
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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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  • 23
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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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  • 24
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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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  • 25
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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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  • 26
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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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  • 27
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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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  • 28
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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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  • 29
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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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  • 30
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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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    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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  • 32
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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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  • 33
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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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  • 34
    ISSN: 0006-3592
    Keywords: acidic fibroblast growth factor ; Escherichia coli ; sterilization ; comparative reasoning tools ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of various medium sterilization conditions on fermentations of a recombinant, acidic fibroblast growth factor (aFGF) producing Escherichia coli have been studied. Changes in the medium resulting from sterilization were monitored by pH and absorption spectra. This simple experiment provided excellent data for the demonstration of the usefulness of comparative reasoning tools in order to evaluate the effect of sterilization on fermentation performance. The time profiles of the main parameters (e.g., carbon dioxide evolution rate, dissolved oxygen, pH, and aFGF productivity) were simplified into piecewise contiguous linear segments, each of which was sequentially numbered. The length, position, and slope of each tine were then characterized. Application of the comparative reasoning tools confirmed that separate sterilization of the glucose was necessary for the success of the process, despite adding to the cost and complexity. The comparative data analysis also showed that scaleup with longer sterilization holding and cooling times would not be detrimental to aFGF production. © 1995 John Wiley & Sons, Inc.
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  • 35
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    Biotechnology and Bioengineering 37 (1991), S. 647-660 
    ISSN: 0006-3592
    Keywords: bacterial chemotaxis ; Escherichia coli ; motility, random ; diffusion chamber assay ; mathematical model ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Bacterial chemotaxis, the directed movement of a cell population in response to a chemical gradient, plays a critical role in the distribution and dynamic interaction of bacterial populations in nonmixed systems. Therefore, in order to make reliable predictions about the migratory behavior of bacteria within the environment, a quantitative characterization of the chemotactic response in terms of intrinsic cell properties is needed.The design of the stopped-flow diffusion chamber (SFDC) provides a well-characterized chemical gradient and reliable method for measuring bacterial migration behavior. During flow through the chamber, a step change in chemical concentration is imposed on a uniform suspension of bacteria. Once flow is stopped, diffusion causes a transient chemical gradient to develop, and bacteria respond by forming a band of high cell density which travels toward higher concentrations of the attractant. Changes in bacterial spatial distributions observed through light scattering are recorded on photomicrographs during a 10-min period. Computer-aided image analysis converts absorbance of the photographic negatives to a digital representation of bacterial density profiles. A mathematical model (part II) is used to quantitatively characterize these observations in terms of intrinsic cell parameters: a chemotactic sensitivity coefficient, μ0, from the aggregate cell density accumulated in the band and a random motility coefficient, μ, from population dispersion in the absence of a chemical gradient.Using the SFDC assay and an individual-cell-based mathematical model, we successfully determined values for both of these population parameters for Escherichia coli K12 responding to fucose. The values obtained were μ = 1.1 ± 0. 4 × 10-5 cm2/s and χo = 8 ± 3 ± 10-5 cm2/s. We have demonstrated a method capable of determining these parameter values from the now validated mathematical model which will be useful for predicting bacterial migration in application systems.
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  • 36
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    Biotechnology and Bioengineering 38 (1991), S. 296-303 
    ISSN: 0006-3592
    Keywords: ethanol ; genetic engineering ; Escherichia coli ; lignocellulose ; xylose ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The conversion of xylose to ethanol by recombinant Escherichia coli has been investigated in pH-controlled batch fermentations. Chemical and environmental parameters were varied to determine tolerance and to define optimal conditions. Relatively high concentrations of ethanol (56 g/L) were produced from xylose with excellent efficiencies. Volumetric productivities of up to 1.4 g ethanol/L h were obtained. Productivities, yields, and final ethanol concentrations achieved from xylose with recombinant E. coli exceeded the reported values with other organisms. In addition to xylose, all other sugar constituents of biomass (glucose, mannose, arabinose, and galactose) were efficiently converted to ethanol by recombinant E. coli. Unusually low inocula equivalent to 0.033 mg of dry cell weight/L were adequate for batch fermentations. The addition of small amounts of calcium, magnesium, and ferrous ions stimulated fermentation. The inhibitory effects of toxic compounds (salts, furfural, and acetate) which are present in hemicellulose hydrolysates were also examined.
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  • 37
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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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  • 38
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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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  • 39
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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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  • 40
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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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  • 41
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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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  • 42
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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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  • 43
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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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  • 44
    ISSN: 0006-3592
    Keywords: promoter system ; recombinant proteins ; Escherichia coli ; plasmid construct ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A pH-inducible promoter system was characterized and its potential applicability in recombinant protein production was evaluated using a plasmid construct, pSM552-545C-, in which the promoter and activator coding sequences of the cad operon were inserted into the upstream region of a lacZ′ reporter gene. Graded gene expression levels with respect to culture pH between 8.0 and 5.5 were observed and the induction range can be as high as 200-fold. The effects of several cultivation parameters, including pH, temperature, induction cell density, and inoculum size, were systematically examined. The practical application of this expression system to high level production of recombinant proteins was successfully demonstrated using a rich medium, superbroth. An extremely high recombinant protein productivity at a value of approximately 1.4 g/L with a specific expression level as high as 35% of total cellular protein can be obtained in a simple batch cultivation. The behavior of this expression system was further investigated using chemostat cultures. An uncommon relationship between the volumetric or specific recombinant protein activity and the dilution rate, with a maximal activity at a dilution rate of approximately 0.4 h-1was observed. © 1995 John Wiley & Sons, Inc.
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  • 45
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    Biotechnology and Bioengineering 47 (1995), S. 596-608 
    ISSN: 0006-3592
    Keywords: amino acid addition ; protein stability ; protease ; fed-batch ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this work, feeding policies aimed to avoid cellular stress responses as indicated by an increase in ATP-dependent proteolysis are tested. A set of experiments was carried out where glucose, IPTG (inducer), and phenylalanine (rate-limiting precursor) were added gradually in a fed-batch fashion. A significant increase in CAT activity was found compared with pulse-induction. In addition, there was a substantial increase in the rate of CAT synthesis as well as in the final specific CAT activity when phenylatanine and the inducer were added simultaneously. CAT degradation was confirmed through Western blotting analysis. Protease analysis (SDS-GPAGE) indicated lower proteolytic activity for the IPTG and phenylalanine fed-batch cases. GroEL immunoas-says indicated that amplification of stress proteins occurred upon CAT induction. This research impacts the yield of soluble cytoplasmic proteins in Escherichia coli and suggests that metabolically based induction/feeding policies are beneficial. © 1995 John Wiley & Sons Inc.
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  • 46
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    Biotechnology and Bioengineering 48 (1995), S. 303-315 
    ISSN: 0006-3592
    Keywords: Plasmid recovery ; Escherichia coli ; Cell disruption ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Five different mechanical cell disruption processes were evaluated as methods to extract plasmids from bacterial cells. The methods used were sonication, nebulization homogenization, microfluidization, and bead milling. The recovery yields of intact plasmids from the various methods were measured by quantitative gel electrophoresis. Bead milling and microfluidization were found to have the highest potential for large scale extraction with total intact recoveries of over 90% and around 50%, respectively. Other methods resulted in substantial plasmid degradation, with recoveries no greater than 20% of the total intact plasmid. © 1995 John Wiley & Sons, Inc.
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  • 47
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    Biotechnology and Bioengineering 38 (1991), S. 340-352 
    ISSN: 0006-3592
    Keywords: Immobilized cells ; Escherichia coli ; microfluorimetry ; DNA staining ; DNA synthesis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pronounced spatial nonuniformities in cell density, physiology, and activity frequently arise within densely packed immobilized cell supports. For a more fundamental understanding of immobilized cell phenomena, we have developed high-resolution microfluorimetric procedures to analyze local variations in both immobilized cell loading and growth rate. Fluorescent staining of total cellular DNA provides a measure of local biomass density. Actively growing (DNA synthesizing) cells are marked by pulse-labeling newly synthesized DNA with the thymine analog, bromouracil. An immunofluorescent technique allows subsequent detection of spatial variations in DNA synthesis rates. These procedures enable the influence of mass-transfer limitations and other immobilization effects on cell distribution and activity to be readily quantified. We demonstrate this approach through analysis of the patterns of growth of Escherichia coli entrapped within Sr-alginate gel beads. The experimental techniques are potentially applicable to a variety of other aggregate cell systems.
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  • 48
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    Biotechnology and Bioengineering 38 (1991), S. 397-412 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; protein synthesis ; metabolic limitations ; cloned-gene expression ; β-galactosidase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Earlier experiments in our lab investigated the metabolic limitations of cloned-gene expression in bacterial cells (for over-production of β-lactamase). These experiments showed that the steady-state concentration of ribosomal RNA decreased upon plasmid amplification while both the synthesis rate and steady-state β-lactamase mRNA level increased significantly. This appeared to indicate substantial limitation exist within the transnational machinery of the bacterial cell at high copy numbers. To establish the generality of this phenomenon, the impact increasing protein expression from pa plasmid by chemically inducing a strong promoter while maintaining constant copy number has been investigated. A plasmid has been constructed which contains the lacZ gene under control of the tac promoter and contains the parB stability locus to maintain plasmid stability. Using this vector, β-galactosidase expression in chemostat cultures operated at specific growth rates of 0.6 h-1 was induced with IPTG such that enzyme activity was varied over a 460-fold range. When fully induced β-galactosidase protein production represented 14 wt % of total cell protein. As transcription was induced, the synthesis rate of the β-galactosidase mRNA increased 42-fold while the steady-state level of β-galactosidase mRNA increased only fourfold. This indicates stability may play a larger role for β-galactosidase expression with a strong promoter than seen with β-lactamase production in the elevated copy number system. Furthermore, rRNA synthesis rates increased at high expression rates as seen in the copy number experiments. However, unlike the amplified-plasmid system, the steady-state levels of rRNA increased as well. Since the total protein levels closely followed the steady-state level of eRNA, transnational limitations are again suggested for the chemically induced transcription system.
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  • 49
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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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  • 50
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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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  • 51
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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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  • 52
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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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  • 53
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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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  • 54
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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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  • 55
    ISSN: 0006-3592
    Keywords: acetate reduction ; Bacillus subtilis ; Escherichia coli ; cloning ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    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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    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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  • 57
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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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  • 58
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    Biotechnology and Bioengineering 47 (1995), S. 139-146 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; fed-batch ; acetate ; glucose ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A Large bioreactor is an inhomogenous system with concentration gradients which depend on the fluid dynamics and the mass transfer of the reactor, the feeding strategy, the saturation constant, and the cell density. The responses of Escherichia coli cells to short-term oscillations of the carbon/energy substrate in glucose limited fed-batch cultivations were studied in a two-compartment reactor system consisting of a stirred tank reactor (STR) and an aerated plug flow reactor (PFR) as a recycle loop. Short-term glucose excess or starvation in the PFR was simulated by feeding of glucose to the PFR or to the STR alternatively. The cellular response to repeated short-term glucose excess was a transient increase of glucose consumption and acetate formation. But, there was no accumulation of acetate in the culture, because it was consumed in the STR part where the glucose concentration was growth limiting. However, acetate accumulated during the cultivation if the oxygen supply in the PFR was insufficient, causing higher acetate formation. The biomass yield was then negatively influenced, which was also the case if the PFR was used to simulate a glucose starvation zone. The results suggest that short-term heterogeneities influence the cellular physiology and growth, and can be of major importance for the process performance. © 1995 John Wiley & Sons, Inc.
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  • 59
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    Biotechnology and Bioengineering 48 (1995), S. 149-157 
    ISSN: 0006-3592
    Keywords: mass transfer ; Escherichia coli ; cell cultures ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Nutrient depletion and inhibitory end-product accumulation are the major problems for hydrogel-entrapment cell cultures. An electrokinetic technique was developed to enhance intrahydrogel mass transfer to overcome these problems. Escherichia coli cells (ATCC 15224) were entrapped in 3.2-mm-thick potassium-K-carrageenan and agarose hydrogel slabs. With a electric current density of 180A/m2 the cell densities were increased by 140%(from 3.9 to 9.6 dry cell weight [DCW] g/L) in potassium-K-carrageenan and by 80% (from 3.9 to 7.0 DCW g/L) in agarose. A mathematical model taking into account nutrient depletion, inhibitory end-product formation, and cell growth kinetics under facultatively anaerobic conditions was developed to rationalize the overall transport and biological behaviors in the hydrogel. The cell growth in hydrogel was successfully simulated. It is concluded that the augmented transports for glucose and inhibitory end-products accounted for these increases in cell growth. The increase in cell density in potassium-K-carrageenan was due to the enhanced removal of inhibitory end-products (through electroosmosis and electro-phoresis: 80%) and due to the augmented glucose transport (through electroosmosis: 20%). © 1995 John Wiley & Sons, Inc.
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  • 60
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The variation in the behaviour of bulk solids as a result of sudden pressure changes in the surrounding gas phase is of interest in the application of lock hoppers for transfer of bulk solids into pressure vessels. By using such sluice systems, problems occur due to consolidation of the pressurized bulk solids. This paper presents the results of experimental and theoretical research, which allow to describe the influence of several parameters on pressure compensation and the degree of consolidation. With reference to the forces and mechanisms responsible for consolidation, it is possible to describe the different consolidation behaviour of incompressible (limestone) and compressible (hard coal) bulk solids.
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  • 61
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    Chemical Engineering & Technology - CET 14 (1991), S. 65-72 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A study was made of the flow characteristics of large oscillating drops of pure liquid-liquid systems, using a thermostatically-controlled, rising drop column, 50 mm in diameter and 1000 mm in length. Mirrors in the jacket enabled front and side views of drops to be photographed simultaneously. Single drops in the size range 5-10 mm were investigated with both mutually-saturated phases and when the solute was being transferred from the dispersed phase. The systems studied were (1) toluene and acetone (dispersed)-water (continuous), and (2) n-heptane and acetone (dispersed)-water (continuous). Acetone concentrations were varied up to 3.75 kmol/m3. The oscillations of a travelling drop were asymmetrical; therefore, the amplitude cannot be expressed accurately in terms of only two axes. The area change of the drop compared to that of a sphere of equal volume ‘ε’, was shown to represent the amplitude accurately. The periods of droplet oscillation were uniform for the mutually saturated systems of constant physical and flow properties but changed when mass transfer was taking place. The interfacial tension exerted a marked effect on the amplitude, which also depended upon the oscillation frequency. The amplitude changed with droplet size in a similar manner to the terminal velocity, i.e. it increased with increasing size until it reached a maximum, subsequently decreasing less rapidly. The drag coefficient increased with increasing rate of mass transfer from the drop. Correlation of the results and the area eccentricity ‘ε’ by dimensional analysis embracing all possible parameters and physical properties affecting drop oscillation, resulted in the correlation ε = 0.22 Sr0.42 We-0.53 M0.13 with a mean deviation of ± 14%. This will facilitate more accurate prediction of the interfacial area for mass transfer calculations, relating to equipment containing droplets in the oscillating regime.
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  • 62
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    Chemical Engineering & Technology - CET 14 (1991), S. 113-117 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The reduction of nitric oxide with ammonia on iron oxide catalysts has been studied in a continuous-flow recycle reactor using simulated flue gas in the temperature range from 573 to 673 K. NO and HN3 concentrations were varied between 0 and 1000 vpm, O2 and H2O concentrations between 0 and 9 vol.-%, the remainder being nitrogen. In the presence of oxygen, the formulated reaction rate equation describes the measured rates of the main reaction NO + 2/3 NH3 ⇄ 5/6 N2 + H2O. Its form corresponds to the Langmuir-Hinshelwood type. The rate equation well fits the data, which cover the whole industrial temperature and concentration range. In the absence of oxygen, the measured reaction rates can be best described by a power law.
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  • 63
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    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new transport system has been developed which combines vibration of the conveying pipe with amplitudes of the order of fractions of a millimetre with rather low superficial velocities of the carrier gas. This new system was specially designed for gentle transport of fragile particulate materials.
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  • 64
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    Chemical Engineering & Technology - CET 14 (1991), S. 147-161 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this study, we investigated the limits of safe operation for a cooled reactor, operated batchwise. As an example of a single-phase reaction, we studied the decomposition of t-butyl peroxypivalate, a well-known organic peroxide, undergoing self-heating at relatively low temperatures. If sufficiently diluted, it can be supplied to a polymerization process from large, cooled but unstirred vessels. We present a number of extensions to the existing homogeneous explosion theory, namely a practical definition of the critical condition, its calculation, and expressions for the available time before runaway in the case of a supercritical condition, taking into account the effects of natural convection inside the vessel and the reactant conversion. The extensions of the theory were confirmed by adiabatic and non-adiabatic runaway experiments on bench scale, and natural convection cooling experiments with liquids in various packages.
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  • 65
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This contribution deals with the consequences of the decrease in diffusion coefficients at the boundaries of miscibility in fluid systems and its effect on the mechanisms of mass transfer. The liquid-liquid (1-1) extraction involving several components is taken as an example for the discussion of this topic. A short review of Hampe's theory on transport mechanisms in the transition regions of interfaces [1] explains the intrinsic interdependence between mass and momentum transfer. Finally, a discussion of the peculiarities of resistances to mass transfer in boundary layers, based on the changes in molecular transport properties, is presented.
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  • 66
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    Chemical Engineering & Technology - CET 14 (1991), S. 200-208 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The catalytic properties of wide-pore zeolites, namely Ni- and Cu-doped faujastie H-Y and H-mordenite (H-M) are compared on the example of conversion of n-hexane. After thermal pretreatment of ion-exchanged zeolites, reduction with hydrogen (PH2 ≤ 40 bar) leads to a large number of metallic clusters and crystallites as well as ionic species in the zeolites. Notwithstanding this variety, the different types of catalysts can be divided into three classes: classical bifunctional catalysts (for example NiHM, NiHY) with ion exchange degrees 〉 30%, monofunctional catalysts, such as CuHM and CuHY with high copper contents (ion exchange degree 〉 40%) and metal-doped (Ni and Cu) zeolites which, depending on the conditions of pretreatment, act as bifunctional and/or monofunctional compounds and, therefore, show very unstable reaction behaviour. In addition to the extent of reduction, the cation distribution affects the activity and deactivation behaviour and, to a lesser extent, the product selectivity. The reaction mechanism of bifunctional catalysis is similar to that observed in platinum-doped catalysts, while the mechanism of monofunctional catalysis resembles the process on undoped zeolites.
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  • 67
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    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This work investigates the thermodynamically based assumption that diffusion at binodal states of unstable fluid systems vanishes. It is shown experimentally that a scaling law, which describes the fall in diffusion near critical points, may also be applied in an extended form to its abatement at the limits of solubility. These results may profoundly affect the modelling of mass transfer between phases whose bulks exhibit states of saturation.
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  • 68
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    Chemical Engineering & Technology - CET 14 (1991), S. 240-247 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A systematic method of controller design is introduced to determine the overall charcteristic behaviour developed on process pole removal. The time delay compensation is automatically incorporated in the proposed control law. The implemented tuning parameters in the law are confined to the range between 0 and 1 to guarantee the stability of the overall control system. Subsequently, the fixed and adaptive control strategies are implemented to simulate a batch PVC reaction system. The adaptive control scheme provides good, roubust control of this simulated reactor notwithstanding the wide range of operating conditions and non-linear dynamics of the system. However, the fixed control scheme performs well only for a noise-free system. In addition, two limiting control laws, derived from the proposed method, are also used to simulate the reactor. The results indicate that these laws are not suitable for this non-linear reaction system.
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  • 69
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    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article describes the effects which occur during three-component mass transfer in liquid-liquid extraction between saturated and undersaturated phases of the system glycerol-acetone-water. The description is based on experiments in a countercurrent spray column. The physical modelling starts with the balance equations of phase flows, taking into account the variation of concentrations and mass flow rates along the column axis as well as backmixing effects. In addition, recent findings on the mechanisms of mass transfer in saturated and undersaturated phasses, deduced from theoretical and experimental results, were used to formulate mass transfer coefficients. These are valid for the region near the interface of drops in a fluid and are inserted into the balance equations. A comparison between experiments and calculations of the observed phenomena is presented.
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  • 70
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    Chemical Engineering & Technology - CET 14 (1991), S. 275-287 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Those chemical and photochemical characteristics of a photochromic system which are necessary for successful application to flow visualization studies in water are identified. The properties of a number of photochromic and photosensitive systems are discussed, and critically examined, with regard to these requirements.
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  • 71
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    Notes: This contribution demonstrates the possibility of using a thermodynamic description of electrolyte solutions directly for computer simulation of distillation columns. Coefficients are determined for a combination of a solvation model with a local composition equation for the activity coefficient. The use of one set of coefficients yields good results of calculation of vapour pressure, vapour phase composition and caloric data of hydrochloric acid. Solution of material and heat balance equations of distillation columns is achieved on an IBM PC with the above description. Two illustrative examples of an absorption and a rectification calculation are presented
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  • 72
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    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The knowledge of multicomponent adsorption equilibria is of great importance for the industrial design and application of adsorptive purification and separation processes. As the experimental determination of the required data is time-consuming and costly, theoretical help is desirable. This contribution presents and assesses the most common theoretical approaches to the calculation of multicomponent adsorption equilibria. The models are based on the assumption of an adsorbed solution, extension of the volume filling of micropores theory or statistical thermodynamics. These models are applied to different adsorptive/adsorbent systems with ideal and non-ideal equilibrium behaviour. It is established that, at low coverages of adsorbent surfaces, all theories provide good estimates of the adsorbed phase composition and total loading while, at high saturations, no model is completely satisfactory.
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  • 73
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    Chemical Engineering & Technology - CET 14 (1991), S. 347-352 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simplified model has been developed to describe ammonia stripping process in spray towers. The effect of various operating parameters including air flow rate, contact time, droplet size, liquid to gas ratios and tower height on the performance of such systems has been studied in detail. The results obtained for ammonia stripping have been compared with the limited experimental data available in literature. The predicted results agree reasonably well with the reported experimental data.
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  • 74
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    Chemical Engineering & Technology - CET 14 (1991), S. 357-362 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of particle size (layer thickness), BET surface area and pore size distribution on the reactivity of CaO derived from limestone were studied by using a pH-stat and the BET method. Visual analysis was obtained with SEM, and X-ray diffraction was used to determine the crystalline components present. It was found that the pore size distribution, including pore volume and pore surface distribution, affected the sulphatization behaviour. The larger pores (D 〉 50 Å) with their pore volume and pore surface were to some extent responsible for both initial reactivity and ultimate capacity of CaO.
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  • 75
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    Chemical Engineering & Technology - CET 14 (1991), S. 376-378 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Activity coefficients of hydrocarbons and chloroparaffins at infinite dilution in methyl phenyl ether, butyl ether and methyl iso-butyl ketone at 293.15 K have been obtained by the retention-time method in a modified gas chromatograph. The experimental data are compared with predictions of the UNIFAC and MOSCED models. Selectivities for the 1,1,1-trichloroetane-1,2-dichloroetane separation process are reported.
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  • 76
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    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A semi-empirical gas-liquid equilibrium model for the absorption of CO2 in aqueous 3M AMP (2-amino-2-methyl-1-propanol) is presented. It applies to high CO2 loadings (y 〉 0.5) in the temperature range between 20 and 50 °C, and is based on experimental solubility and pH determinations. For a given amine concentration, it yields the equilibrium partial pressure of CO2 as a function of only two variables: the CO2 loading and temperature. The model correlates the expressions for the chemical equilibria involved as follows: pCO2 = m y × 10x, where pCO2 is the equilibrium partial pressure, x = logK - pH, m is the amine molarity, y the CO2 loading, and K is a parameter involving Henry's law constant, H, and the first dissociation constant, K1, of carbonic acid. pH is found to depend on both temperature and CO2 loading while logK depends only on the CO2 loadIng. Correlations for pH and logK are presented. The model fits own data for 3M AMP very well as well as the equilibrium data found in recent literature.
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  • 77
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    Chemical Engineering & Technology - CET 15 (1992), S. 91-98 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The transport of corrosive liquids containing solids, in conventional energy and process technology as, for example, in oil and gas production, places high demands on the selection of materials for plant construction. A discussion on the chemo-mechanical load demonstrates the significance of hydrodynamics in selecting the test method. Choosing a pipe with an abrupt expansion of cross-section as a flow model yields a new method of simulating operation conditions. By subjecting ferrous materials used for hydraulic machinery to representative media from three important industries, a broad spectrum of results can be obtained. Classification of the results leads to four basic types of mass loss behaviour in materials as functions of flow rate. These findings yield guidelines for the selection of materials and the dimensioning of flow equipment.
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    Chemical Engineering & Technology - CET 15 (1992), S. 124-130 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The adsorption of p-nitrophenol (PNP) from an NaOH, H3PO4-buffered, aqueous solution on activated carbon was studied at pH 2, 7, and 8.5. Comparative studies were then carried out on ozonized carbon and ozonized PNP solution. In the latter case, PNP adsorption was performed from an ozonized solution, and also after exposing the activated carbon to the ozonized PNP solution for 24 h. At acid pH, PNP adsorption was found to be markedly higher than at neutral and weakly basic pH's. Ozonization of carbon affects the results only at pH 7 and 8.5. Ozonization of PNP led to a small decrease in adsorption, irrespective of the method of contect between carbon and ozonization reaction products. The rate was faster at acid than at basic pH, and the ozonization speeded up the adsorption slightly but only at pH 8.5. Application of a mathematical model indicated that mass transfer in bulk solution controls the rate of adsorption.
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    Chemical Engineering & Technology - CET 15 (1992), S. 162-167 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new model is derived to establish the effective nucleation rate in terms of the MSMPR-theory. The model is based on the concept of two-step crystal growth, secondary nucleation, nuclei activation and metastable supersaturation. A comparison between model prdictions and experimental data from different crystallizers is presented. This model, which integrates hydrodynamics directly into its theory, proves to be extremely suitable for describing the MSMPR-crystallization process.
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    Chemical Engineering & Technology - CET 15 (1992), S. 193-201 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A laboratory fixed-bed reaction column is presented, in which a heterogeneouslyl catalyzed reversible fluid-phase reaction and the separation of the reaction product from unreacted or inert components by distillation are performed simultaneously. The rection studied, the formation of methyl tertiary butyl ether (MTBE) from methanol and isobutylene, is catalyzed by strongly acidic macroreticular ion exchange resins. Packing elements are Raschig rings, manufactured for the first time from the above-mentioned ion exchange resins. This in-house made catalyst shows practically the same ion exchange capacity as the commercial products. The control system installed in the equipment is capable of correcting strong disturbances os the steady state. As shown by expwrimental results, the superimposed distillation process occurs in such a way that the chemical equilibrium of the MTBE synthesis does not limit isobutylene conversion.
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  • 81
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    Chemical Engineering & Technology - CET 16 (1993), S. 24-25 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Measurements of surface tension of flame-, plasma- and UV-treated PP-EPDM samples show that all three methods of pretreatment significantly increase the surface tension. The flame or plasma pretreatment increases especially the polar element of surface tension while the UV-treatment enhances the disperse element. The measurements were carried out with a new fully automatic unit for determining contact angles (AURAM).
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    Chemical Engineering & Technology - CET 16 (1993), S. 35-39 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Experimental data are reported for ion exchange equilibria of binary systems on Amberlite IR-120 resin, at different temperatures and total ionic concentrations in aqueous solutions. These systems exhibit non-ideal behaviour in both phases, and the equilibrium characterization has been based on the law of mass action, using Wilson and Meisssner, and Kusik equations to correlate the activity coefficients in the resin and aqueous solutions, respectively. Equilibrium constants, standard thermodynamic properties (Gibbs free enthalpy changes, enthalpy and entropy) and Wilson binary parameters were obtained as functions of temperature.
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    Chemical Engineering & Technology - CET 16 (1993) 
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    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
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    Chemical Engineering & Technology - CET 16 (1993), S. 62-67 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The kinetics of methane oxidative coupling (OCM) was studied using 1 g of Na-Mn2O3 catalyst at 1073 to 1123 K, in an integral flow reactor (I.d. = 10 mm), at atmospheric pressure with methane and oxygen partial pressures of 0.27 and 0.13 bar, respectively, so that the ratio of CH4 to O2 was 2. The flow rate range was 50 to 200 ml/min. the kinetic data were analyzed by the Rideal-redox type of rate equation assuming the methyl radical and active surface oxygen to be the steady-state intermediates. Oxidation and reduction rate constants (Kox, Kred) for methane consumption were calculated from experimental catalysis results by computer simulation using the multiple least squares method. The activation energies at rate constants Kox and Kred for this type of catalyst were reported as 43.26 and 62.2 kcal/mol, respectively.
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    Chemical Engineering & Technology - CET 15 (1992), S. 69-81 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The photochemical reaction SiHCl3+Cl2→SiCl4+HCl is of industrial importance in the production of pure silicon for the semiconductor industry. The present paper deals with an experimental investigation of the kinetics of this purification step. The main reaction occurs in the liquid phase and is initiated by light. In this gas-liquid reaction system, all components, which are present in both phases, were analyzed by gas chromatography. Reaction progress was monitored by recording the pressure rise. Apart from the influence of wavelength and radiation intensity on the above photoreaction, reactions in the absence of light and on the wall were also investigated.
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    Chemical Engineering & Technology - CET 15 (1992), S. 114-118 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A bubble column absorber was used to investigate kinetics of the reaction between carbon dioxide and aqueous solutions of diisopropanolamine (DIPA), by means of gas absorption experiments. These were conducted in the temperature range of 20 to 40°C, with DIPA concentrations from 5 to 500 mol/m3, and CO2 partial pressures between 5 and 101 kPa. A model based on the Danckwerts' surface reneval theory was used to analyze the experimental results and to determine the rate constant. The obtained data support the assumption of a second-order overall reaction, with the rate constants being well correlated by the Arrhenius equation: \documentclass{article}\pagestyle{empty}\begin{document}$$ k_2 = 1.112 \times 10^{10} \exp \left({ - {{4848} \mathord{\left/ {\vphantom {{4848} T}} \right. \kern-\nulldelimiterspace} T}} \right).$$\end{document}
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    Chemical Engineering & Technology - CET 16 (1993), S. 130-146 
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    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new set of rate equations for gas desorption has been derived from the laws of thermodynamics and motion and the kinetic theory of gases. The equations assume that gas desorption or absorption originally represents an isothermal change at the interface, similarly to condensation or vaporization. The mass transfer in gas desorption or absorption is initially caused by external work being done on the system and takes place as work done by the system which instantly develops a concentration gradient, increases in internal energy and temperature, and a deviation from the ideal gas behaviour inside MTZ in the process of mass transfer. After cessation of external work, these conditions disappear following the transfer of a small residual mass at a constant velocity and a decrease in internal energy inside the MTZ.
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    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Theoretical and experimental investigations were performed with cold water models of bottom-blown metallurgical ladles in order to develop a mathematical description of the mixing processes in such reactors. This analytical model is based on the subdivision of the reactor into interacting subspaces, whereby each subspace is regarded as an ideal mixer. The convective mass transfer between the subspaces is represented by constant coupling coefficients, and the mass balance for the material to be mixed in provides a homogeneous system of differential equations to determine the time dependence of concentration in the individual subspaces. Different combinations of interacting subspaces are represented. The flow field and the distribution of turbulence intensity measured by LDA in water models and the concentration profiles in mercury models measured by EMF cells show that a subdivision into five subspaces is adequate. Size, position and interaction coefficients are also determined by experiments. The calculated mixing times agree well with those measured.
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  • 89
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    Chemical Engineering & Technology - CET 16 (1993), S. 200-205 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Henry's law is often used to correlate the solubility of a gas in water. Such an approach is frequently employed to calculate the solubility of hydrogen sulphide in water. However, Wright and Maass [1] clearly demonstrated that the strict Henry's law did not apply to the solubility of hydrogen sulphide in water. That is, the solubility of hydrogen sulphide is not directly proportional to its partial pressure, even at seemingly low pressures. By accounting for the non-idealities in the vapour phase, Carroll and Mather [2] showed how a simple modification of Henry's law could be used for moderate pressures (up to 1 MPa). In this paper, a further extension of Henry's law is used to model the vapour-liquid equilibrium at higher pressures and temperatures. It is also applied to liquid-liquid and vapour-liquid-liquid equilibria, areas where Henry's law is rarely used.
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  • 90
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    Chemical Engineering & Technology - CET 15 (1992), S. 269-277 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The chemical and photochemical properties of a photochromic water soluble triaryl methane dye sulphite, acid magenta monomethyl magenta sulphite, (AMMM · SO3)4-, have been examined, and the application of this photochromic system to the visualization of water flows is discussed. Addition of sulphite to an aqueous solution of the parent dye acid magenta monomethyl magenta, AMMM2-, (a methylated derivative of acid magenta, C.I. acid violet 19) leads to formation of the colourless sulphite complex, (AMMM · SO3)4-. Pulsed irradiation of a solution of this compound, with light of wavelength shorter than ≈ 340 nm, causes transient production of the parent dye, whose lifetime is influenced by pH, sulphite concentration, temperature and ionic strength. Longest useful lifetimes are of the order of a few seconds and a general equation is presented for the calculation of dye lifetimes for a variety of conditions. The complex formation constant for (AMMM · SO3)4- is 1.8 (± 0.2) × 105 M-1, while that for the weakly photochromic hydroxide (AMMM · OH)3- is 1.3 (± 0.3) × 106 M-1. The quantum yield of dye production at 308 nm is ≈ 0.5 - 1.0, and this is discussed along with the absorption spectra of AMMM2- and (AMMM · SO3)4- with respect to the penetration characteristics of a pulse of 308 nm laser radiation with an energy of a few millijoules. The action spectrum for dye production from (AMMM · SO3)4- suggests that there is a non-photochromic absorption band at about 350 nm, an assumption which is supported by the failure to detect efficient dye production when using 355 nm excitation from a Nd/YAG laser.
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  • 91
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    Weinheim : Wiley-Blackwell
    Chemical Engineering & Technology - CET 15 (1992), S. 300-312 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Thick-walled components subjected to pulsating internal pressure are widely applied in high-pressure technology and in manufacturing processes such as fluid-jet cutting and high-pressure cleaning, mainly in conjunction with reciprocating pumps. Corrosive fluids require high-strength and tough chrome-nickel steels with soft martensitic or semi-austenitic structure. This contribution reports on the fatigue of thick-walled plain and cross-bored pipes made from high alloy chrome-nickel steels such as X5 CrNiMoCu 21 8 and X5 CrNiMo 16 5. The speciments, uniaxial standard form and thick-walled pipes, were cut from forged blocks in the three axial directions. For loading with pulsating pressure, a suitable, high-frequency piston pulsation machine has been developed. The fatigue tests on pipe specimens show typical Woehler characteristics with only slight scatter and relatively good isotropy. The surprisingly large admissible pulsating pressure can be explained for the applied steels by dynamic generation of residual stresses as a result of shake-down effects. Presentation in a Smith digram explains the occuring dynamic shake-down and its favourable results in comparison to the more brittle highly tensile steels. It also reveals that heat treatment to higher tensile strength does not always yield an increase in the admissible pulsating pressure. It will be shown that static autofretting and shake-down affect the fatigue strength of thick-walled pipe specimens in the same way. Tests with internal liners in the tube specimens provide indications on the sensitivity of material failures towards fluids. The investigation aids the understanding of the fatigue behaviour and the design of components made of modern high-strength corrosion resistant steels.
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  • 92
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    Chemical Engineering & Technology - CET 16 (1993), S. 213-217 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Treatment of a given multicomponent cationic mixture was studied applying the emulsion liquid membrane (ELM) separation technique for Zn recovery. Experimental apparatus and conditions are described. Comparative studies using two zinc-selective ion exchangers (thio-DEHPA and dithio-DEHPA) were carried out whereby the composition of the liquid membrane and the operating conditions were investigated for both extractants. A useful method of representing the concentrations of the feed and the receiving aqueous phase is proposed, which allows the occurrence of break-up and/or osmosis to be easily detected.
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  • 93
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    Chemical Engineering & Technology - CET 16 (1993), S. 238-242 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This contribution presents a method of evaluating electrical conductivities of pure organic liquids. Emphasis is placed on the methodology of selection, correlation and analysis of experimental data. The proposed method brings order out of conflicting, confusing and chaotic experimental observations. It is also shown that the consistent and evaluated data are generated at a fraction of the cost and time required for producing the original raw experimental data.
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  • 94
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    Weinheim : Wiley-Blackwell
    Chemical Engineering & Technology - CET 16 (1993) 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
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  • 95
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Based on the dispersion model rather than on the conventional pluf-flow model, a method of predicting the transient behaviour of parallel and counterflow heat exchangers is developed, whereby the effects of shell-side flow maldistribution and the finite heat capacities of both fluids, tube bundle and shell are considered. The final solution has been obtained by numerical inversion of the Laplace transform, to determine the transient responses to arbitrary inlet temperature variations. The experiment was carried out in a one pass tube heat exchanger. The comparison between therotical and experimental results is presented and the feasibility of this method is discussed.
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  • 96
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    Chemical Engineering & Technology - CET 16 (1993), S. 413-421 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This contribution reviews the models for equilibrium and kinetics of the system Zn/D2EHPA/n-dodecane (and other diluents), published during the past decade, as well as values of the appropriate model parameters, i.e. of equilibrium and reaction rate constants. The reviewed kinetic models comprise those for kinetic, diffusional and mixed mass transfer regimes. It is shown that, under certain conditions, the rate equation within the kinetic regime has the same mathematical structure as the equations for the overall mass transfer rate within the diffusional and mixed mass transfer regimes. This means that the same equation interprets experimental data from each of the three regimes. Thus, a fit of experimental data to a model is not sufficient for discriminating between the mass transfer regimes. Results of kinetic experiments, using a stirred mass transfer cell with constant interfacial area, are presented.
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  • 97
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The separation of ethoxylated aliphatic amines from zinc containing effluents with macroporous polymeric adsorbents Wofatit EP 61, EP 62, Y 59, Y 77 and active carbons has been examined. The adsorption behaviour has been investigated as a function of temperature, time and concentration of Na2SO4, ZnSO4 and H2SO4. The macroporous polymers EP 61, Y 77 and Y 59 are capable of separating ethoxylated aliphatic amines from effluents of viscose fibre industry at higher temperatures and under weakly acidic conditions. In contrast to active carbon, inorganic components are not adsorbed on the investigated macroporous resins. Data obtained in a concentration range of 0 to 10 g of adsorbate per litre produced a Langmuir profile for adsorption of ethoxylated aliphatic amines by polymeric adsorbents.
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  • 98
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The present work investigates the energy consumption of a solids production process. Such a process consists of three different operation units: crystallization/precipitation, solid-liquid separation by centrifuges or filters and drying of wet crystals. Each unit is analyzed with the objective of minimizing the energy consumption. Operating conditions, process design and other characteristic parameters (crystal size, porosity and physical proporties) are discussed with the aim to evaluate their influence on the process.
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  • 99
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    Weinheim : Wiley-Blackwell
    Chemical Engineering & Technology - CET 16 (1993), S. 102-108 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Power consumption under non-aerated and aerated conditions was measured in a 0.45 m diameter tank containing a range of xanthan gum solutions, using the Scaba 6SRGT impeller. Well-mixed caverns were measured in a transparent Carbopol solution. The data are compared with previous results obtained with a Rushton turbine in the range of Reynolds numbers, Re, between 1 and 2000. In each case, the non-aerated power numbers were similarly related to Re except that the Po values of the Scaba 6SRGT were 3 to 4 times smaller. Under aerated conditions, the aeration rate does not affect the power number, Pog, with the actual value falling as a result of increasing xanthan concentration. The Pog/Po vs Re curve passes through a minimum which is lower at higher xanthan concentrations. At the same Reynolds number, the decline in power consumption of the Scaba agitator due to aeration is 20 to 50% smaller than that observed with a Rushton turbine. Well-mixed cavern shapes and sizes are also very similar to those observed with Rushton turbines and can be predicted from equations available in literature. However, instabilities in torque could present a problem in broths containing xanthan in concentrations above 35 kg m-3. Nevertheless, the Scaba agitator offers an interesting retrofitting possibility for xanthan fermentation, which could lead to improved performance.
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  • 100
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    Weinheim : Wiley-Blackwell
    Chemical Engineering & Technology - CET 16 (1993) 
    ISSN: 0930-7516
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
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