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  • 201
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991), S. 673-680 
    ISSN: 0006-3592
    Keywords: indole alkaloid production ; Catharanthus roseus ; Agrobacterium rhizogenes ; hairy root cultures ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Catharanthus roseus hairy root cultures, genetically transformed with Agrobacterium rhizogenes, produce a wide variety of indole alkaloids. The effect of sucrose, phosphate, nitrate, and ammonia concentrations on growth and indole alkaloid production of C. roseus hairy root cultures were studied by using statistical experimental designs and linear regression analysis. Contradictory effects of these nutrients on growth and indole alkaloid production were found. The maximal growth was obtained by having 77. 8 mg NaH2PO4 · H2O/L and 1. 311 g KNO3/L in the medium, whereas the specific production of alkaloids was highest at the lowest levels of all the nutrients studied. The maximal dry weight was obtained with high values of sucrose and ammonia, but clear optimum concentrations could not be found. When having enough nutrients to support reasonable growth, it appeared difficult to affect the specific alkaloid production rates considerably. The growth (dry wt.) with the optimized nutrient concentrations in the medium was more than 50% better than in the control medium with about the same alkaloid production.
    Additional Material: 7 Ill.
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  • 202
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 203
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991), S. 703-707 
    ISSN: 0006-3592
    Keywords: Dextransucrase ; Leuconostoc mesenteroides ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: High yields of the enzyme dextransucrase have been produced repeatedly by fed-batch fermentation techniques. Activities in excess of 21.9 U/cm3 have been obtained by culturing Leuconostoc mesenteroides NRRL B-512(F) under nonaerated fed-batch fermentation conditions. Aerobic fermentations carried out under identical conditions have consistently produced enzyme of less than 17 U/cm3, but with no difference in the final cell concentration in the broth. Different types of yeast extract have been found to have significant effect on the final cell concentration and more especially on the enzyme activity with enzyme yields varying by as much as 50% when different types of yeast extracts were used.
    Additional Material: 4 Ill.
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  • 204
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991), S. 729-735 
    ISSN: 0006-3592
    Keywords: amperometric biosensor ; hypoxanthine ; electropolymerization ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An amperometric biosensor for hypoxanthine was constructed by forming a layer of crosslinked xanthine oxidase on a platinum electrode, followed by electropolymerization of a submonolayer film of resorcinol and para-diaminobenzene. The fabricated electrodes were evaluated for speed of response, sensitivity, and reusability. Optimal performance was obtained with enzyme-based electrodes sparsely covered with film which was formed by electropolymerization in less than 6 min. The resulting electrodes exhibited linear response to hypoxanthine in the. range 5-300 μM with a response time of 2 min. Application of the biosensor in monitoring hypoxanthine content of fish extracts yielded results which agreed well with spectrophotometric assays using soluble xanthine oxidase. The biosensor was stable for 60 days when stored at 4°C in phosphate buffer and it could be used continuously for 6 h with over 50 assays.
    Additional Material: 7 Ill.
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  • 205
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991), S. 762-769 
    ISSN: 0006-3592
    Keywords: animal cell culture ; bioreactor ; airlift ; fiber-bed bioreactor ; oxygen transfer ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simple hydrodynamic model is introduced to describe the airlift fiber-bed bioreactor, which can enhance the volumetric productivity of anchorage-dependent animal cell cultures. By applying the model, liquid flow rates and volumetric mass transfer coefficients are predicted and are in agreement with experimental measurements. Consequently, the optimal reactor configuration giving the maximal oxygen supply is derived. Also, theoretical scaleup potential of this concentric internal loop reactor is considered for volumes ranging from 10 to 67,000 L with which cell densities of 5.1 × 107 and 1.2 × 107 cells/cm3, respectively, can be maintained.
    Additional Material: 8 Ill.
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  • 206
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 196-205 
    ISSN: 0006-3592
    Keywords: maximum principle ; Bang-bang control ; extended Kalman filter ; PF system ; glutathione production from yeast ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The optimal of the specific growth rate was obtained with simple mathematical model in a yeast fed-batch cultures. The model was based on the mass balance around the fed-batch system and the relationship between the specific growth rate, μ, and the specific production rate of glutathione, ρG. The optimal profile of μ was calculated as a bang-bang type, That is μ, should start from the maximum value, μmax and should be kept at μmax; then μ should be switched to μc, which gives a maximum value of ρG. It was proven from the maximum principle that switching was needed only once, with the switching time from μmax to μc depending on the final required glutathione content. Finally, this ideal profile of μ for the maximum production of glutathione was realized by manipulating the substrates feed rate in the fed-batch culture. Using the extended Kalman filter and a programmed-controller/feedback-compensator (PF) system, μ could be controlled at the optimal profile obtained. As a result, the maximum production of glutathione was accomplished fairly successfully. However, further improvement in the controller performance for μ is desired. The control strategy employed here can be applied to other batch reaction processes.
    Additional Material: 12 Ill.
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  • 207
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 217-223 
    ISSN: 0006-3592
    Keywords: sucrose ; biosensor ; Zymomonas mobilis ; interfering compounds ; invertase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new biosensor for specific determination of sucrose was developed using an oxidoreductase of Zymomonas mobilis and invertase. Cells of Z. mobilis were permeabilized with toluene in order to utilize the enzymes of glucose-fructose oxidoreductase and gluconolactonase inside the intact cells. Permeabilized cells and invertase were coimmobilized in a gelatin membrane, and a whole cell enzyme electrode was constructed by fixing the membrane on a pH electrode. The production of hydrogen ion was detected using the biosensor-connected microcomputer, and the concentration of sucrose was determined by using both the initial rate and the steady-state methods. Optimum conditions for biosensor response were pH 6.2 and temperature 35°C. The effect of interfering compounds on the electrode response was investigated, and the interference by various sugars was eliminated by determining sucrose concentration using the steady-state method. The biosensor developed is simple and reproducible, and the calibration curve for sucrose is linear up to 70 g/L.
    Additional Material: 10 Ill.
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  • 208
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991), S. 869-875 
    ISSN: 0006-3592
    Keywords: scu-PA ; pro-urokinase ; yeast ; respiratory quotient ; fermentation ; Saccharomyces cerevisiae ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Secretion of a nonglycosylated form of human pro-urokinase, also known as single-chain urinary plasminogen activator (scu-PA), from Saccharomyces cerevisiae is described. A “supersecreting” yeast strain harboring multiple copies of integrated plasmids was grown batchwise and at constant respiratory quotient (RQ) in 20-L fermenters. Because the promoters used to drive expression of the pro-urokinase genes are not tightly regulated, secretion into the culture supernatant was growth associated. Although the final cell density achieved in the perturbed-batch fermentation (45 g dry wt/L) was less than that observed in the RQ-controlled culture (77 g dry wt/L), the scu-PA titer in the perturbed-batch fermentation (1863 IU/mL) was nearly twice that attained at constant RQ (1108 IU/mL). The effects on cell growth and scu-PA titer of other process variables (pH, temperature, phosphate concentration, and medium composition) are also discussed.
    Additional Material: 5 Ill.
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  • 209
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 280-295 
    ISSN: 0006-3592
    Keywords: AtT-20 cells ; secretory proteins ; intracellular trafficking ; conversion compartment ; trans-Golgi ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: After their synthesis, secretory proteins in animal cells undergo a series of transport and processing steps before they are secreted. The amount and quality of protein obtained in culture medium depends on the rates of these intracellular steps. We present a model of recombinant protein trafficking in mouse pituitary AtT-20 cells based on currently available biological knowledge, plausible hypotheses, and quantitative secretion results, and we use it to simulate the dynamics of basal and induced secretion and to predict the dynamics of intracellular trafficking events. Besides the endoplasmic reticulum and Golgi, the model recognizes a conversion compartment (CC) where final processing of protein occurs, a storage compartment from which protein is secreted only in the presence of secretion stimulus, and constitutive and pseudoregulated (PR) pathways of secretion. The model further assumes that the protein flux is split between CC and PR and that the storage compartment exerts a negative feedback on protein flux through CC. The model predictions are compared with experimental results on secretion of human growth hormone (hGH) and insulin related peptides and on accumulation of hGH upon removal of secretion stimulus. The model is in agreement with data when either of two hypotheses is implemented: (a) cells always exhibit a high sorting efficiency at the trans-Golgi, but CC has the capacity to process only a fraction of the protein flux leaving the Golgi compartment; (b) the processing capacity of CC never becomes saturated, but significant missorting at the trans-Golgi occurs; in the case, the protein flux toward the plasma membrane becomes split both at the trans-Golgi cisternae and between CC and PR. The usefulness of the type of model considered in providing a quantitative description of intracellular events and in designing new experiments is discussed.
    Additional Material: 9 Ill.
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  • 210
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    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.
    Additional Material: 3 Ill.
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  • 211
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 331-339 
    ISSN: 0006-3592
    Keywords: Thalictrum rugosum ; alkaloid production ; berberine ; bioreactor ; airlift ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Airlift bioreactor operations have been studied for the growth-associated production of secondary metabolites from plant cell suspension cultures. The model system used in this work was Thalictrum rugosum producing berberine, an isoquinoline alkaloid. The airlift system was well suited for growth of Thalictrum cell suspension cultures unless the cell density was high. At high cell density, the airlift system with a draught tube was not adequate due to large aggregates clogging the recirculation paths. This was overcome by use of a cell scraper in the reactor. For berberine production, gas-stripping also played a significant role and it was discovered that CO2 and ethylene were important for product formation. By supplying a mixture of CO2 and ethylene into the airlift system, the specific berberine content was increased twofold. It is evident that continuous gas sparging was harmful for the production of berberine without supplementation with other gases.
    Additional Material: 6 Ill.
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  • 212
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 371-379 
    ISSN: 0006-3592
    Keywords: plant tissue culture ; Papaver somniferum ; linear growth ; phosphate limitation ; modeling ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In examining the growth kinetics of cell suspensions of opium poppy (Papaver somniferum), the increase in biomass with time was observed to be linear over the entire batch growth period of up to 20 days. Although batch growth profiles were reproducible utilizing the same inoculum, growth rates varied tremendously when experiments were inoculated with cells from different flasks. Both of these phenomena are difficult to explain with conventional batch growth models. In a series of a experiments, phosphate was determined to be the growth-rate-limiting substrate. By expressing growth rate in terms of the intracellular reserves of phosphorus, a growth model which expresses kinetics in terms of the intracellular phosphorus contents of the cells is shown to predict both linear growth character and inoculum dependent variability in growth. The stationary phase phosphate content of seven plant suspension cultures of different plant species was found to be comparable to phosphorus levels of phosphate-starved poppy cells, which suggests that phosphate limitation may be common for plant tissue culture. The applicability of this model to other biological systems which display similar batch growth patterns when subjected to inorganic nutrient deprivation is discussed.
    Additional Material: 5 Ill.
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  • 213
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 159-168 
    ISSN: 0006-3592
    Keywords: spin filter ; plugging ; fouling ; perfusion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Mouse-mouse hybridomas (15 μm mean diameter) were cultivated in a simulated perfusion reactor with spin filter and external recirculation of the medium. Proteins at high concentrations, such as 10% foetal calf serum (FCS), were found to be not responsible by themselves for fouling, even at high recirculation rates. Stainless steel (10 μm pores) in contrast to polyamide (11 μm proes) led to a great accumulation of dead cells and nucleic acids on the screen, finally leading to fouling, as shown by biochemical and microscopic examinations. It is suggested that the high surface charge density of metals compared to polyamide is responsible for attachment of various residues. Stainless steel should rather be replaced by a resistant and nontoxic synthetic material, such as polyamide 66 which was successfully used. FCS should be avoided, since it seems to increased the fouling phenomenon. Moreover, the pore size of the screen should be carefully defined according to the wide size distribution of living and dead cells of the line used (33% of variation of the mean size in our case) as well as fragments. The purpose of the screen being to get rid of fragments and small dead cells, and not to wash too many new small cells, a good retention was achieved here by a 10-μm opening.
    Additional Material: 6 Ill.
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  • 214
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 869-876 
    ISSN: 0006-3592
    Keywords: fermentation ; pervaporation ; immobilized yeast ; S. cerevisiae ; ethanol ; membrane ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A system comprised of an immobilized yeast reactor producing ethanol, with a membrane pervaporation module for continuously removing and concentrating the produced ethanol, was developed. The combined system consisted of two integrated circulation loops: In one the sugar-containing medium is circulated through the membrane pervaporation module. The two loops were interconnected in a way allowing for separate parameter optimization (e.g., flow rate, temperature, pH) for each loop.The fermentation unit was 2.0 L bioreactor with five equal segments, packed with 5-mm beads of immobilized yeasts. The bead matrix was a crosslinked polyacrylamide hydrazide gel coated with calcium alginate. The fast circulation loop of the bioreactor allowed for efficient liberation of CO2 at the top of the immobilized yeast reactor. Continuous operation of the uncoupled reactor for over 50 days with inflowing defined medium or dilute molasses at a residence time of 1.25 h yielded ethanol at a rate of about 10 g/L h.The pervaporation unit was constructed from four 60-cm-long tubular membranes of silicone composite on a polysulfone support. The output from the fermentor was circulated through the inside of the tubes of a unit with a total surface area of 800 cm2, having an average flux of 150 mL/h, and selectivities to ethanol vs. water up to 7. A vacuum of 30 mb was applied to the outside of the tubes, removing 20-30 g of ethanol per hour, which was collected in condensors. The continuous removal of ethanol, avoiding inhibition of the fermentation process, resulted in an improved productivity and allowed the use of high sugar concentrations (40% wt/vol) offering the potential of a compact system with reduced stillage.The combined system of ethanol production and removal enabled an operative steady state at which the liquid volume of the system, and the concentrations of ethanol within the reactor (˜4% wt/vol), as well as within the flux crossing the pervaporation membrane (17%-20% wt/vol) were kept constant. At the steady state, a 40% wt/vol sugar solution could be continuously added to the fermentor when 12%-20% wt/vol clear ethanol solution was continuously removed by the pervaporation unit. Membrane fouling was reversed by short washing steps, and continuous step operation was maintained by working with two different modules that were interchanged. In this manner, long term continuous operation (over 40 days) was achieved with a productivity of 20-30 g/L h, representing over a twofold increase relative to the continuously operated reactor uncoupled from the membrane and a fivefold increase in comparison with the value obtained fro a corresponding batch fermentation.
    Additional Material: 8 Ill.
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  • 215
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 923-928 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Significant quantities of heavy metals will adsorb onto modified bone gelatin beads. As this adsorption occurs, the bead can undergo a substantial volume change. Research has shown that the equilibrium bead diameter was a function of the solution pH and the ion concentration in the solution. Here, we demonstrate that under certain conditions, the volume of the beads that absorbed the metal was only 35% of the bead volume when no metal was adsorbed. By taking advantages of these size changes, a fluidized-bed separator can be operated such that natural segregation of loaded beads occurs. This phenomenon may facilitate the design of continous separators for the recovery and concentration of heavy-metal-contaminated waters. These concepts are demonstrated using Cu2+ adsorption onto such beads.
    Additional Material: 6 Ill.
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  • 216
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 948-951 
    ISSN: 0006-3592
    Keywords: amperometric method ; oxygen consumption ; reaction mechanism ; colorimetric method ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simple amperometric method has been developed for the determination of amino groups on a solid support. The method is based on oxygen consumption during the reaction between the bifunctional reagent gluaraldehyde and the amino groups on the solid support. Oxygen consumption was monitored with a Clark oxygen electrode. AHSepharose 4B, Affi-Gel 102, and Amino-Celluofine were used as amino-terminal solid supports. The concentration of amino groups on each support was calculated from a calibration curve of the standard compound. When hexamethylenediamine was used as the standard, the concentration of amino groups on AH-Sepharose 4B and Affi-Gel 102 by the present method showed good agreement with that by the conventional method. In the case of Amino-Cellulofine, ethanolamine had to be selected as the standard. The relative standard deviation for six successive determinations of AH-Sepharose 4B was 3.2%; the polymerization of glutaraldehyde showed no effect on the result. The spectral property of AH-Sepharose 4B treated with glutataraldehyde showed no effect on the result. The spectral property of AH-Sepharose 4B treated with lutaraldehyde suggested that the reaction mechanism of the present method was based on the oxygen consumption during the formation of the pyridinium salt on the support. A comparative study of the present method with a colorimetric method with Traut's regent is also included.
    Additional Material: 4 Ill.
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  • 217
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 961-961 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 218
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 986-994 
    ISSN: 0006-3592
    Keywords: protein partitioning ; polyethyleneglycol/dextran systems ; isoelectric point ; polymer molecular weight ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We report the partition coefficient, Kp′ at the isoelectric point of lysozyme, chymotrypsinogen A, albumin, transferrin, and catalase in 64 different polyethylene(PEG)/ dextran(Dx)/water systems. We study the trends of the partition coefficient with protein type, polymer concentration, and polymer molecular weight. We find that the partition coefficient decreases with increasing tie line length for lysozyme, albumin, transferrin, and catalase for which Kp is less than 1, but increases for chymotrysinogen for which Kp is larger than 1. The effect of the tie line length on the partition coefficient is larger for the large proteins than for the small proteins. The partition coefficient decreases with increasing protein molecular weight except for lysozyme suggesting that lysozyme is present as a dimer or a trimer. The partition coefficient decreases with increasing PEG molecular weight, but the magnitude of the increase is larger for the smaller PEG molecular eights and tends to level of at high PEG molecular weight. The partition coefficient increases with increasing dextran (Dx) molecular weight for chymotrypsinogen but decreases for catalase. The partition coefficients of lysozyme, albumin, and transferrin increase with increasing Dx molecular weight from Dx 104 to Dx 1.1 × 105 and then slightly decrease from Dx 1.1 × 105 to Dx 5 × 105. The experimental results are analyzed using a statistical thermodynamics model. The experimental results are analyzed using a statistical thermodynamics model. The experiments suggest that protein partitioning at the isoelectric point in aqueous two-phase systems is strongly related to the size of the proteins and polymers. Finally, the impossibility of obtaining data completely independent of polymer concentration is emphasized.
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  • 219
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 1012-1019 
    ISSN: 0006-3592
    Keywords: invertase ; polyelectrolytes ; polyampholytes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In connection with our work on polyelectrolyte complex formation with polyampholytes, the interaction between invertase and several linear polyelectorlytes has been investigated by means of turbidimetry, light scattering measurements, and determination of the enzyme activity. Polyelectrolyte complex formation of invertase was shown to occur with cationic polyelectrolytes only. The light-scattering data yield information on aggregation and desegregation processes in complex formation. As indicated by our results, only a part of the protein molecules is engaged in this Coulombic interaction, and this part shows a rather small enzyme activity only. Thus, a direct interaction between invertase and a cationic polyelectrolyte is no effective approach to enzyme binding, but a complete immobilization of invertase can be achieved via an “inclusion flocculation” with a symplex formed by interaction between an anionic and a cationic linear polyelectrolyte or via immobilization in symplex microcapsules.
    Additional Material: 8 Ill.
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  • 220
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 1041-1049 
    ISSN: 0006-3592
    Keywords: diffusion ; lactose ; get matrix ; immobilization ; growing cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Effective diffusion coefficients (De) of lactose in κ-carrageenan (2.75% wt/wt)/locust bean gum (0.25% wt/wt) (LBG) gel beads (1.5-2.0-mm diameter)with or without entrapped lactic acid bacteria (LAB) were determined at 40°C. The effects of lactose concentration, bacteria strain (Streptococcus salivarius subsp. thermophilus and Lactobacillus casei subsp. casei) and cell content at various steps of the fermentation process (after immobilization, pre-incubation of the beads and successive fermentations) were measured on De as a first step for process modelling. Results were obtained from transiend concentration changes n well-stirred lactose solutions in which the beads were suspended. A mathematical model of unsteady-state diffusion in a sphere was used, and De was obtained from the best fit of the experimental data. Diffusivity of lactose in cell-tree beads was significantly lower than in pure water mainly because of the obstruction effect of the polymer chains and the hydration region. Furthermore, effective diffusivity and equilibrium partition factor were independent of lactose concentration in the range from 12.5 to 50 g/L. No significant difference was found for De (effective diffusivity) and Kp (partition) coefficients between beads entrapping S. thermophilus (approximately 5 × 109 CFU/mL) and cell-free beads. On the other hand higher cell counts obtained with L. casei (close to 1.8 × 1011 CFU/mL) increased mass transfer resistance resulting in lower effective diffusivities and Kp. Finally, the effects of the type of bacteria and their distribution in the beads on the diffusivity were also discussed.
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  • 221
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    Biotechnology and Bioengineering 38 (1991), S. 1082-1090 
    ISSN: 0006-3592
    Keywords: fermentation ; recombinant fermentation ; cyclic fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A two-stage, cyclic fed-batch fermentation process to produce recombinant human lymphokine was designed. The organism used in the study was Escherichia coli K-12 containing a temperature-sensitive walkaway plasmid bearing an insert which codes for a human lymphokine. Transcription of the recombinant gene is controlled by a lambda repressor/pL promoter system. The lambda promoter is regulated by the temperature-sensitive product of the cl857 gene at 30°C, but at 42°C the promoter is derepressed. The first or growth, stage of the process was maintained at 28°C and operated in the fed-batch mode. The vessel was fed at a rate which gives a constant specific growth rate using a media designed to maintain a constant optical density OD600 of 50. After the volume in the first stage reached the maximum working volume of the vessel (12 L), a portion of the vessel contents was transferred to the second stage. The second, or induction/product formation, stage also operated in the fed-batch mode, was kept at 42°C, and was fed with a media that is conducive to recombinant human lymphokine synthesis. An optical density of more than 100 was consistently achieved in the second stage. Thirty cycles were completed with a consistent yield of human lymphokine and cell density in each cycle. The process was used to produce 200 L of OD600 50 material from the first stage in 10 days. The volumetric productivity (g lymphokine/L. day) of the two-stage, cyclic fed-batch process is twice that of a single-stage, fed-batch fermentation process.
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  • 222
    ISSN: 0006-3592
    Keywords: ABE fermentation ; Clostridium acetobutylicum ; whey permeate ; packed bed reactor ; pervaporation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An integrated solvent (ABE) fermentation and product removal process was investigated. A stable solvent productivity of 3.5 g/L h was achieved by using cells of Clostridium acetobutylicum immobilized onto a packed bed of bonechar, coupled with continuous product removal by pervaporation. Using a concentrated feed solution containing lactose at 130g/L, a lactose value of 97.9% was observed. The integrated fermentation and product removal system, with recycling of the treated fermentor effluent containing only low amount of solvents (/but lactose and acids), leads to only low acid losses. Therefore, most of the acids are converted to solvents, and this results in a high solvent yield of 0.39 g solvents/g lactose utilized. The pervaporation system provided a high product removal rate even at low solvent concentrations. A solvent membrane flux of 7.1 g/m2 h with a selectivity of 5 was achieved during these investigations. The system proved to be very reliable.
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  • 223
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    Biotechnology and Bioengineering 38 (1991), S. 1166-1172 
    ISSN: 0006-3592
    Keywords: hollow-fiber capillary reactor ; L-alanine dehydrogenase ; NAD regeneration ; elementary reaction kinetics ; multistage stirred tank reactor model ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Continuous production of L-alanine with conjugated enzyme systems of alanine dehydrogenase (AlaDH) and lactate dehydrogenase (LDH) or alcohol dehydrogenase (ADH) was carried out with NAD regeneration in an ultrafiltration hollow-fiber capillary reactor (HFCR) which was proposed as a test bioreactor with very small scale. In the AlaDH/LDH system, pyruvate is the intermediate product for L-alanine so that an optimal point existed in pyruvate concentration for the production rate of L-alanine. NAD cycling number of 4850 and L-alanine productivity of 61.7 mmol/L h were obtained at the best condition. In the AlaDH/ADH system, however, the substrate inhibition in the AlaDH reaction by pyruvate should be considered and the best results of NAD cycling number and L-alanine productivity were 2700 and 13.5 mmol/L h, respectively. In consideration of concentration distribution and mixing in the axial direction on an HFCR, performance of the reactor was theoretically analyzed with a multistage stirred tank reactor model combined with the kinetic model based on all the elementary reactions involved. Although quantitative discrepancy existed in some cases, the present theoretical model could explain experimental results and is expected to be generally applicable to standard hollow fiber reactors.
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  • 224
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    Biotechnology and Bioengineering 38 (1991), S. 1203-1209 
    ISSN: 0006-3592
    Keywords: oxygen fluctuations ; cephamycin C ; antibiotics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An experimental Monte Carlo method was used to study the effect of fluctuations in oxygen concentration on the synthesis of antibiotics by Streptomyces clavuligerus. Air was supplied to the culture in a 2-L fermentor in random cycles following the lognormal distribution in order to model the circulation within large production-scale vessels. Each cycle consisted of air supply for 5 s followed by no aeration for the balance of the cycle time which ranged from 8 to 44 s, with a mean time of 20 s. Comparable experiments were also conducted with constant period cycling of air and with continuous supply of air. The yields of cephamycin C and its precursor, penicillin N, were suppressed by the Monte Carlo simulation of circulation in a large tank, as compared to constant period cycling. The concentration of dissolved oxygen remained at a low, ca. 5% of saturation, for 5-10 h longer during the Monte Carlo experiment than during the periodic aeration experiment. The biosynthetic enzymes, which are sensitive to oxygen levels, were likely affected not only by the mean time of cycling but also by the distribution of the cycles.
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  • 225
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    Biotechnology and Bioengineering 38 (1991), S. 1233-1238 
    ISSN: 0006-3592
    Keywords: membrane bioreactor ; gas transfer ; pulsatile flow ; external oxygenation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We show the design features of a membrane bioreactor based on pulsatile flow across dimpled membranes. Results show an enhanced mass transfer of air of at least five-fold magnitude as compared with flat membranes. An increased working volume form 20 mL to 120 mL reduced the kLA at a given Reynolds number because of axial mixing of fluid from the deoxygenated end chamber. The bioreactor was used to supply air to a hybridoma mammalian cell line, and the calculated oxygen uptake showed that high-density cultures could be maintained in a 20mL, single-dimpled cultures could be maintained in a 20 mL, single-dimpled membrane system. Indirect aeration of a 2 L continuous stirred tank reactor, by a double-membrane system, showed that air could be supplied to mammalian cells at cell densities of approximately 4 × 106 /mL.
    Additional Material: 7 Ill.
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  • 226
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    Biotechnology and Bioengineering 38 (1991), S. 1285-1291 
    ISSN: 0006-3592
    Keywords: Tripterygium Wilfordii ; plant cell culture ; suspension ; medium ; immobilization ; bioreactor culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The plant Tripterygium wilfordii produces di- and triterpenes of interest for male contraception and treatment of arthritis and skin disorders. Cell line TRP4a obtained form this plant in 1981 was reported to produce these valuable compounds at yields (∼0.04% of the biomass dry weight) higher than found in the plant (0.001%). In order to improve this production, studies were carried out to determine the feasibility of eliminating the troublesome component of coconut milk originally used to culture this cell line. A defined formulation suitable for growth ad maintenance has been developed. This medium consisted of Gamborg's PRL4 or B5 medium supplemented with 2 mg L-1 2,4-dichlorophenoxyacetic acid and 20 g L-1 sucrose. Furthermore, monitoring of carbohydrate uptake revealed that T. wilfordii cells, contrary to many plant cell species, did not hydrolyze sucrose extra-cellularly before uptake. Replacement of this disaccharide by glucose or fructose increased specific growth rate from 0.15 to 0.25 day-1. As tripdiolide is reported to be present in broth extract in significant amounts, plant cell immobilization technology offers a promising alternative to suspension cultures, especially in view to on line harvesting of the product. Surface immobilized T. wilfordii cell cultures were successfully carried out in 2-L bioreactors. Their biomass production and carbohydrate uptake were comparable to those observed for shake flask grown suspension cultures. Higher nitrate and ammonium uptake were found in immobilized cultures.
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  • 227
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    Biotechnology and Bioengineering 38 (1991), S. 1318-1324 
    ISSN: 0006-3592
    Keywords: glucose metabolism ; pet operon ; E. coli ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The glucose metabolism of an Escherichia coli strain bearing mutations abolishing both acetyl phosphotransferase (PTA) and acetate kinase (ACK) activities was studied under aerobic and anaerobic conditions. These studies were conducted in a complex medium with the mutant carrying no plasmid, the mutant carrying the common cloning vector pUC19, and the mutant carrying a plasmid bearing the “pet” operon that encodes Zymomonas mobilis pyruvate decarboxylase and alcohol dehydrogenase activities. The mutant carrying no plasmid showed lower specific growth and glucose uptake rates relative to the parent wild-type strain (K-12), Lactic acid was produced at higher levels than the wild type, and considerable amounts of pyruvic acid were secreted as an unusual byproduct. Analysis of other fermentation products showed low but significant amounts of acetic acid, no accumulation of formic acid, and lower secretion of succinate and ethanol. The maintenance of the plasmid pUC19 in the mutant negatively affected metabolism. Expression of the pet operon overcame the metabolic stress caused by the plasmid, enhancing growth and glucose uptake rates to the values observed in the plasmidfree mutant. Also, expression of the pet operon allowed consumption of pyruvate accumulated during the first hours of fermentation.
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  • 228
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    Biotechnology and Bioengineering 38 (1991), S. 1353-1363 
    ISSN: 0006-3592
    Keywords: Klebsiella pneumoniae ; metabolic regulation ; continuous cultures ; cybernetic model ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The transient behavior of a continuous culture of Klebsiella pneumoniae with mixed feed of glucose and xylose arising from step-up and step-down in dilution rates and from a feed-switching experiment is presented. he organism gradually switches from simultaneous utilization of the substrates at low growth rates to preferred utilization of the faster substrate (i.e, supporting a higher growth rate) at high dilution rates. The metabolic lags following a step increase in dilution rate and a significant accumulation of the slower substrate during the transient period result from the effects of metabolic regulation. The cybernetic modeling approach that successfully described the foregoing situations with single-substrate feeds is employed to describe mixed substrate behavior. The parameters in the mixed-substrate (glucose and xylose) model are the same as those in the single-substrate models with the singular exception of the rate constant for the xylose growth enzyme synthesis. The reason for this discrepancy is discussed in detail. It appears that the constitutive rate of enzyme synthesis for growth on a given substrate may be related to the past history of the organism in regard to whether or not the organism has been exposed to the particular substrate. Thus, the results further demonstrate the ability of the framework to effectively describe metabolic regulation in batch, fedbatch, and continuous microbial cultures.
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  • 229
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    Biotechnology and Bioengineering 38 (1991), S. 653-658 
    ISSN: 0006-3592
    Keywords: light irradiation ; anthocyanin production ; Perilla frutescens ; cell culture ; bioreactor cultivation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: After a series of experiments on photoperiodicity and light intensity under daylight supplied by an ordinary fluorescent lamp in cultivations using a flask and a roux bottle, it was found that irradiation at 27.2 W/m2 for the whole period was effective for anthocyanin production by a suspended culture of Perilla frutescens (shiso). A high amount of anthocyanin pigments, 3.0 g/L, was obtained in a bubble column bioreactor after 10 days of cultivation at an aeration rate of 0.1 vvm with light irradiation at 27.2 W/m2, while 2 g/L was obtained at 13.6 W/m2 and very little at 54.4 W/m2. A high amount of anthocyanin pigments, 2.9 g/L, was also produced using an aerated and agitated bioreactor at an agitation speed of 130 rpm, an aeration rate of 0.1 vvm and light irradiation intensity of 27.2 W/m2. The amount of anthocyanin produced was more than twice that without light irradiation, Keeping the other cultivation conditions the same. The results obtained also showed that the amount of anthocyanin pigment accumulated in a shake flask could be rather well reproduced in bioreactors for both aerated culture, and aerated and agitated culture, by improving the conditions of light irradiation, which conspicuously affects metabolite formation.
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  • 230
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    Biotechnology and Bioengineering 38 (1991), S. 679-687 
    ISSN: 0006-3592
    Keywords: mathematical models ; cross regulation ; repressor synthesis control ; gene expression ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Molecular-level mathematical models have been used to evaluate the effectiveness of eight different configurations of repressor synthesis control on the regulation of cloned gene expression initiated from a promoter-operator sequence. Both single and dual-repressor situations were considered, employing genetically structured models for the lac and λPR promoter-operators in example calculations. Simulation results suggest that the most effective mode of cloned gene expression control is a cross-regulation configuration carried on a multicopy plasmid. This system was able to control cloned gene transcription in the uninduced state over a broad range of plasmid copy number and also provided the highest overall transcription rate in the induced state. The general strategies suggested by these simulations should be applicable for other repressor-operator-promoter systems in diverse hosts.
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  • 231
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    Biotechnology and Bioengineering 38 (1991), S. 727-732 
    ISSN: 0006-3592
    Keywords: lipase kinetics ; Candida cylindracea ; hydrolysis of triacetin ; hollow-fiber membrane reactor ; immobilization ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The aptitude of a hollow-fiber membrane reactor to determine lipase kinetics was investigated using the hydrolysis of triacetin catalyzed by lipase from Canadida cylindracea as a model system. The binding of the lipase to the membrane appears not to be very specific (surface adsorption), and probably its conformation is hardly altered by immobilization, resulting in an activity comparable to that of the enzyme in its native form. The reaction kinetics defined on the membrane surface area were found to obey Michaelis-Menten kinetics. The specific activity of the lipase in the membrane reactor was found to be significantly higher than in an emulsion reactor. The activity and stability of the enzyme immobilized on a hydrophilic membrane surface seem not to be influenced significantly by the choice of the membrane material. The hollow-fiber membrane reactor is a suitable tool to assess lipase kinetics in a fast and convenient way.
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  • 232
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    Biotechnology and Bioengineering 38 (1991), S. 1001-1006 
    ISSN: 0006-3592
    Keywords: subtilisin ; enzymes ; inactivation ; stabilization ; organic solvents ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The stability of the serine proteases from Bacillus amyloliquefaciens (subtillisin BPN') and Bacillus licheniformis (subtilisin Carlsberg) was investigated in various anhydrous solvents at 45°C. The half-life of subtilisin BPN' in dimethyl-formamide dramatically depends on the pH of the aqueous solutions from which the enzyme was lyophilized, increasing from 48 min to 20 h when the pH is raised from 6.0 to 7.9. Both subtilisins exhibited substantial inactivation during multihour incubations in tert-amyl alcohol and acetonitrile when enzymatic activities were also measured in these solvents; however, when the enzymes were assayed in water instead, hardly any loss of activity was detected. This surprising difference appears to stem from the partitioning of the bound water essential for catalytic activity from the enzymes into the solvents. When assayed in organic solvents, this time-dependent stripping of water results in decay of enzymatic activity; however, when assayed in water, where the dehydrated subtilisins can undergo rehydration thereby recovering catalytic activity, little inactivation is observed. In agreement with this hypothesis, the addition of small quantities of water tert-amyl alcohol stabilized the subtilisins in it even when enzymatic activity was measured in the nonaqueous solvent. Ester substrates (vinyl butyrate and trichloroethyl butyrate) greatly enhanced the stability of both subtilisins in organic solvents possibly because of the formation of the acyl-enzymes.
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  • 233
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    Biotechnology and Bioengineering 38 (1991), S. 1034-1040 
    ISSN: 0006-3592
    Keywords: amino acid fermentation ; culture redox potential ; fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We investigated the relationship of dissolved oxygen and culture redox potential (CRP) on amino acid production. Corynebacterium glutamicum ATCC 14296 was used for all experiments. The fermentation can be divided into a growth phase and a production phase. Our results indicate that in order to get higher amino acid production, a lower oxygen supply during the exponential phase is favored. A higher oxygen supply rate appears to be necessary during the production phase. Culture redox potential (CRP) was used to monitor the fermentation. CRP readings were observed to drop to a characteristic minimum value as the metabolic state changed from a growth to production phase. This was evidenced by the commencement of amino acid production and a simultaneous uptake of lactate. Upon lactate exhaustion, the CRP increased abruptly. At the same time, maximal amino acid yields were observed. By the use of minimum CRP as an indication of metabolic phase changes, the agitation rate was changed to increase oxygen supply during the production phase. This significantly increased amino acid production. These results show that culture redox potential measurements can be used to monitor and optimize amino acid production by process manipulation.
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  • 234
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    Biotechnology and Bioengineering 38 (1991), S. 1059-1064 
    ISSN: 0006-3592
    Keywords: Amino acid extraction ; di(2-ethylhexyl)phosphoric acid ; liquid membrane ; L-lysine ; pertraction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Kinetics of liquid membrane (Pertraction) recovery of L-lysine from dilute aqueous solutions is studied in a tow-compartment glass cell. A 5% (vol) solution of the cation exchange carrier di(2-ethylhexyl)phosphoric acid in n-decane was used as intermediate, membrane liquid. The third stripping phase was 1/v hydrochloric acid. The reaction mechanism and stoichiometry were defined, and on the basis of the proposed mathematical model of the process and the experimental data obtained, the mass transfer coefficients were evaluated. It was found that overall transfer rate is controlled by the eddy diffusion of transported species in the donor and membrane liquids. The results proved the feasibility of the pertraction process for recovery and concentration of L-lysine from its dilute aqueous solutions.
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  • 235
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    Biotechnology and Bioengineering 38 (1991), S. 1100-1109 
    ISSN: 0006-3592
    Keywords: cephamycin ; Nocardia lactamdurans ; nutrient regulation ; antibiotic fermentation ; nitrogen metabolism ; phosphate limitation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A High cephamycin C producing strain of Nocardia lactam-durans was used to study cell growth and antibiotics production in defined media. Batch fermentations in shake flasks and stirred tanks showed that antibiotic production occurred during cell growth and the production rate rapidly decline as the growth slowed. Glutamate served as a primary substrate during this phase. Later, ammonia was utilized along with a remainder of the glucose. Rapid antibiotic production occurred in this phase. Increased glutamate promoted higher growth, a rise in ammonium ion concentration, and a marked reduction in antibiotic titers. An increase of the glucose concentration along with the glutamate concentration balanced to the medium; no ammonium ion rise occurred and a peak specific antibiotic titer comparable to the control medium was obtained. In a phosphate-limited medium, cell growth equivalent to the control medium and increased antibiotic titers were obtained. In these experiments, adjustment of Na+ and K+ ion concentration equal to that in the control medium was found to be important. Based on carbon and nitrogen balances, the activity of the key nitrogen metabolism enzymes, and the published literature, a two-stage model of regulation is suggested.
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  • 236
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    Biotechnology and Bioengineering 38 (1991), S. 1119-1130 
    ISSN: 0006-3592
    Keywords: continuous fermentation ; on-line analysis ; biomass production ; microfluorometry ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A process for the continuous fermentation of the genetically modified, nitrogenase-producing Escherichia coli C-M74 (pUS1)-strain has been developed. This strain, which is able to fix molecular nitrogen, has the nifgenes of the bacterium Klebsiella pneumoniae. Cell growth and nitrogenase activity of the enzyme have been optimized both in batch and continuous fermentations. For the fermentations, trial runs were performed by cultivating the E. coli cells in 50-ml culture bottles. The medium composition was varied in order to provide high biomass production and nitrogenase activity. For an effective fermentation control, an on-line analysis was built up for the substrates ammonium and glucose. Other medium components such as ampicillin, citric acid, acetic acid, nitrogenase activity, and protein were measured by using different off-line methods. Modern optical methods like in-line microfluorometry for monitoring the culture fluorescence and laser flow cytometry for the estimation of DNA and protein content were also employed. Plasmid stability was also determined.
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  • 237
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    Biotechnology and Bioengineering 38 (1991), S. 1153-1158 
    ISSN: 0006-3592
    Keywords: chemical stabilization of enzymes ; thermal in activation of enzymes ; thermal equivalence ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Acid phosphatase thermal deactivation follows a complex path: an initial decay toward an equilibrium distribution of at least two intermediate structures, mutually at the equilibrium, followed by a final breakdown toward a completely inactive enzyme configuration. The results obtained in the presence of sorbitol have been compared to those produced in the course of purely thermal deactivation of the native enzyme. For any sobitol concentration, an equivalent temperature is calculated that results in exactly the same activity-versus-time profile. This suggests enzyme deactivation to be controlled by a single, unchanging step. Immobilized enzyme runs have been performed, as well, by entrapping acid phosphates within a polymeric network formed onto the upstream surface of an ultrafiltration membrane. The stabilizing effect of entrapment cumulates with that produced by sorbitol. In this case, however, an equivalent temperature cannot be determined, thus indicating that a different deactivation mechanism is followed.
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  • 238
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    Biotechnology and Bioengineering 38 (1991), S. 1182-1189 
    ISSN: 0006-3592
    Keywords: photobioreactor ; ultrfiltration ; photoautotrophic ; DNA histograms ; cell cycle ; flow cytometry ; chlorella vulgaris ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A photobioreactor system has been designed, constructed and implemented to achieve high photosynthetic rates in high-density photoautotrophic algal cell suspensions. This unit is designed for efficient oxygen and biomass production rates, and it also can be used for the production of secreted products. A fiber-optic based optical transmission system that is coupled to an internal light distribution system illuminates the culture volume uniformly, at light intensities of 1.7 mW/cm2 over a specific surface area of 3.2 cm2/cm3. Uniform light distribution is achieved throughout the reactor without interfering with the flow pattern required to keep the cells in suspension. An on-line ultrafiltration unit exchanges spent with fresh medium, and its use results in very high cell densities, up to 109 cells/mL [3% (w/v)] for eukaryotic green alga chlorella vulgaris. DNA histograms obtained form flow cytometric analysis reveal that on-line ultrafiltration influences the growth pattern. Prior to ultrafiltration the cells seem to have at a particular point in the cell cycle where they contain multiple chromosomal equivalents. Following ultrafiltration, these cells divide, and the new cells are committed to division so that cell growth resumes. The Prototype photobioreactor system was operated both in batch and in continuous mode for over 2 months. The measured oxygen production rate of 4-6 mmol/L culture h under continuous operation is consistent with the predicted performance of the unit for the provided light intensity.
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  • 239
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    Biotechnology and Bioengineering 38 (1991), S. 1259-1259 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 3 Ill.
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  • 240
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    Biotechnology and Bioengineering 38 (1991), S. 1065-1081 
    ISSN: 0006-3592
    Keywords: mRNA ; transcription ; translation ; excretion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A predictive, simple, structured model describing the synthesis of α-amylase by Bacillus amyloliquefaciens was formulated. Three key intracellular processes were identified (i.e, translation, and excretion) along with two key intracellular components (i.e., mRNA and the intracellular form of the α-amylase enzyme). Nearly all the model parameters were estimated by means of performing independent experiments, primarily fed-batch experiments. The model was shown to predict transient system behavior in batch and in fed-batch operation with some limitation and minor model parameter revisions. Since a principal objective was to demonstrate that independent experimental parameter determination can be used to construct the predictive model, further fine-tuning of the parameters may be necessary before application for optimization and control purposes.
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  • 241
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    Biotechnology and Bioengineering 37 (1991), S. 505-511 
    ISSN: 0006-3592
    Keywords: perfluorocarbon emulsions ; oxygen permeability ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of interfacial surfactant molecules on oxygen transfer through oil/water phase boundary has been studied in FlurO2TM emulsions, i.e., perfluorocarbon (PFC) emulsions developed as oxygen carriers in cell culture. Measurements of oxygen permeability were made with a polarographic oxygen electrode in pure PFCs and in emulsions with various PFC volume fractions. Comparison of the experimental results with the theoretically derived values of relative oxygen permeability clearly indicates that the mass transfer resistance caused by the interfacial surfactant layer in PFC emulsions is insignificant. Therefore, oxygen dissolved in the enclosed PFC phase is readily available to cells growing in the aqueous media and FlurO2 emulsions with very fine emulsion particles (〈 0.2 μm) can be used to effectively enhance gas/liquid interfacial oxygen transfer in bioreactors. The inadequacy in describing mass transfer in heterogeneous systems, such as the PFC emulsions, by conventional concentration-based oxygen diffusion coefficients has also been discussed.
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    Biotechnology and Bioengineering 37 (1991), S. 537-543 
    ISSN: 0006-3592
    Keywords: colloidal particles ; γ-globulin ; adsorption ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of surface properties on the adsorption of bovine γ-globulin, a model protein for antibody, was studied. Polystyrene latex (PS), hydrophilic copolymer lattices of styrene/2-hydroxyethyl methacrylate [P(S/HEMA)], styrene/ methacrylic acid [P(S/MAA)] and methyl methacrylate/ 2-hydroxyethyl methacrylate [P(MMA/HEMA)], and colloidal silica were used. The adsorption isotherms of γ-globulin on these colloidal particles were measured as a function of pH and ionic strength. The hydrophilic particles showed low affinities for γ-globulin at alkaline pH, while PS showed high affinities for γ-globulin over the whole range of pH and ionic strength. The γ-globulin adsorption on hydrophilic particles was highly reversible with respect to the pH and ionic strength compared with that on PS. These differences indicate that the dominant driving forces of adsorption are related to the hydrophilicity of particles. The adsorption isotherms of all colloidal particles showed the plateau values, and the order of maximum values of plateau adsorption was P(S/MAA) 〉 PS or P(S/HEMA), silica 〉 P(MMA/HEMA). Thus, they were also affected by the charged groups and the hydrophilicity of the surfaces. On the other hand, the plateau values of all colloidal particles were more or less symmetrical with a maximum at around the isoelectric point of γ-globulin at an ionic strength of 0.01. This behavior is attributed to the important role of the lateral interaction between the adsorbed molecules at low ionic strength.
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  • 243
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    Biotechnology and Bioengineering 37 (1991), S. 587-590 
    ISSN: 0006-3592
    Keywords: Saccharomyces ; lactose fermentation ; glucose fermentation ; reverse osmosis membrane ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel concept of membrane bioreactor in which living cells are sandwiched between ultrafiltration (UF) and reverse osmosis (RO) membranes was applied for lactose fermentation to ethanol by genetically engineered yeast cells. The productivity of the Lactophile 13B strains was higher than that of the Lactophile 13D strains. In both cases performance data similar to those for glucose fermentation to ethanol by Saccharomyces cerevisiae were obtained. However, the operational stability of recombinant yeast cells was improved in the new bioreactor in comparison to the stability of these cells in a shake flask.
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  • 244
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    Biotechnology and Bioengineering 37 (1991), S. 591-595 
    ISSN: 0006-3592
    Keywords: tylosin ; macrocin ; dissolved oxygen ; aeration rate ; Streptomyces ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Different dissolved oxygen concentrations and aeration rates were imposed on a stable mutant of Streptomyces fradiae during the antibiotic-producing phase. At high aeration rate (1 vvm), the tylosin yield in the fermentor broth with dissolved oxygen (DO) concentrations controlled close to 100% saturation (6-8 ppm) increased 10% as against uncontrolled. The rates of cellular growth, oil consumption, and tylosin production were severely reduced when DO concentration fell below 25% saturation, but all resumed to their initial rates when DO was raised to saturation level again. The DO concentration in combination with air flow rate affected the pattern of the antibiotics produced. At high DO levels, an additional macrolide antibiotic, macrocin, was synthesized to more than one-third the amount of tylosin at high aeration rate (1 vvm). On the other hand, tylosin production rate remained constant and no significant amount of macrocin was produced at low aeration rate (0.2 vvm).
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  • 245
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    Biotechnology and Bioengineering 37 (1991), S. 627-638 
    ISSN: 0006-3592
    Keywords: solubility parameters ; hydrophobicity index ; Hansen parameter ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Widespread commercial application of enzymes as catalysts for specialty or commodity chemical synthesis will require their use in nonaqueous systems. While a number of non-aqueous enzyme applications have been demonstrated, the lack of useful rules for predicting enzyme-solvent interactions has hindered the development of this technology. Both Hildebrand and solvent hydrophobicity (octanol-water partition coefficient) parameters have been used previously to correlate and predict enzyme activity in nonaqueous systems, with some success, but any single-parameter approach is inherently limited in its ability to reflect the spectrum of possible enzyme-solvent interactions. Therefore, this study evaluates the three-dimensional solubility parameter space, as proposed by Hansen, to correlate and predict enzyme activity in microaqueous, miscible, and biphasic nonaqueous systems. Preliminary results suggest that Hansen parameters may be useful for correlating nonaqueous enzyme activity, and that the dispersive and polar parameters may be disproportionately important in single-phase microaqueous systems. The Hansen hydrogen-bonding parameter appears to be the only parameter yet evaluated capable of correlating the water requirement for enzyme activity in microaqueous systems, suggesting that water affects protein structure through enthalpic rather than entropic processes in nonaqueous systems. Insufficient data are available for miscible and biphasic systems, but it is proposed that enzyme activity may correlate with the average solubility parameters of miscible systems and of the aqueous phase in biphasic systems.
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  • 246
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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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  • 247
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    Biotechnology and Bioengineering 37 (1991), S. 693-695 
    ISSN: 0006-3592
    Keywords: insulin ; single chain des-(B-30)-insulin precursor ; lysyl endopeptidase ; des-(B-30)-procine insulin ; proinsulin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: It has been shown that the single-chain des-(B-30)-insulin precursor (SCI) can be converted into human insulin ester by transpeptidation using trypsin in the presence of a threonine derivative. The present study demonstrates that Achromobacter lyticus protease 1 (lysyl endopeptidase) can catalyze the transpeptidation reaction more efficiently than can trypsin. It is also shown that des-(B-30)-insulin (DAI) can be produced by hydrolysis of SCI with the lysyl endopeptidase. Since it is well known that SCI can be produced by gene technology, the following method is recommended for industrial production of human insulin ester: hydrolysis of SCI with lysyl endopeptidase followed by coupling of the resulting DAI with a threonine derivative using trypsin or lysyl endopeptidase.
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  • 248
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    Biotechnology and Bioengineering 37 (1991), S. 708-715 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A mathematical model for the growth of a single cell of E. coli on medium containing amino acid is presented. A mixture of purified amino acids (glutamate, aspartate, serine, tyrosine, and leucine) combined in the ratios found in a natural digest (casein) were employed as the nitrogen source. Each of these amino acids is the representative of a different family of amino acids. The transport mechanisms and assimilation routes for each amino acid were inserted into the prototype model. The enzyme activities and saturation constants used in the model were based on literature data. The maximum velocities for uptake systems were calculated from experimental data. The formation and homeostasis of amino acid pools were regulated through cross-control of the activities of biosynthetic enzymes and of membrane transport of exogenous nutrients. The size of each amino acid pool was determined with mass balance equations that included terms for a transport system, a biosynthesis system, a transaminase enzyme system for interchange between the amino acid families, and a consumption system. The predictions of the extended model with regard to nutrient concentrations and growth rates compared well with the experimental data.
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  • 249
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    Biotechnology and Bioengineering 37 (1991), S. 736-745 
    ISSN: 0006-3592
    Keywords: computer image analysis ; electrophoresis patterns ; DNA manipulations ; recombinant Escherichia coli ; metabolic analysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Relative levels of many individual proteins in Escherichia coli HB101 strains with 0, 37, 56, and 240 plasmids per chromosome were determined by computer image analysis of two-dimensional gel electrophoresis patterns. The plasmids investigated had very similar sequences except for small domains encoding the represser of plasmid replication. At the intermediate plasmid copy number of 56, levels of several of the TCA cycle enzymes (oxoglutarate dehydrogenase complex, succinate thiokinase, and succinate dehydrogenase) as well as in aspartate transcarbamoylase increased. At a plasmid copy number of 240, higher amounts of PEP carboxylase as well as several of the heat shock proteins were observed. Furthermore, at high plasmid levels, significant decreases occurred in growth rate, pyruvate kinase I, pyruvate dehydrogenase complex, unadenylated glutamine synthetase, aspartate transcarbamoylase as well as in several of the proteins involved in translation. Decreases in ribosome content as well as in the free 30S and 50S ribosomal subunit pool fractions were also observed in separate analyses. These results indicate that recombinant DNA manipulations can cause major alterations in numerous host cell properties which could significantly influence cloned protein production or metabolic engineering endeavors.
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  • 250
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    Biotechnology and Bioengineering 37 (1991), S. 770-777 
    ISSN: 0006-3592
    Keywords: laccase ; Trametes versicolor ; Fomes fomentarius ; affinity chromatography ; purification ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A rapid and convenient method for graduation, isolation, and purification of laccase from Trametes versicolor and Fomes fomentarius culture fluids was developed. For purification affinity chromatography on syringyl- and vanillyl-controlled porosity glass (CPG) columns was applied. The purified laccase of F. fomentarius was immobilized on porous glass. Some properties of the immobilized enzyme in comparison to the free one are discussed.
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  • 251
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    Biotechnology and Bioengineering 37 (1991), S. 1076-1086 
    ISSN: 0006-3592
    Keywords: plasmids-mathematical models ; multimerization ; segregational instability ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of plasmid multimerization on segregational instability was investigated using a structured, segregated model of genetically modified Escherichia coli cells. By including the multimerization of plasmids, the model can predict the proportion of each multimer in the total plasmid population. Simulation results suggest that the plasmid copy number is controlled by the total plasmid content (i.e., total number of plasmid origins) in the host cell and that multimerization reduces the total number of independent, monomeric segregation units. However, multimerization is found to have a minor effect on decreasing plasmid segregational stability for multicopy plasmids with average copy number per cell greater than about 25. Also model predictions were used to test whether or not a nonrandom plasmid distribution at cell fission could cause segregational instability. Even in the case of severely biased partitioning, plasmids whose copy number is above 45 per cell do not show significant segregational instability. The results suggest that when the ColE1-type plasmid does not encode and express any large or disruptive foreign proteins, the copy number of 45 per cell may be the threshold at which only growth rate-dependent instability is responsible for overall plasmid instability.
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  • 252
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    Biotechnology and Bioengineering 38 (1991), S. 1-10 
    ISSN: 0006-3592
    Keywords: lactic acid fermentation ; fermentation ; microbial fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Most fermentation models presented in the literature are unstructured, i.e., the biomass composition is assumed constant during all operating conditions. These models are unable to simulate experiments carried out at widely different operating conditions. It is therefore interesting to examine simple structured models where knowledge of the cell physiology is taken into account in the modeling phase. In this article, a simple structured model is presented. The model is based on experimental work with the lactic acid bacteria Streptococcus cremoris, but due to the similarities in basic metabolism for many microorganisms it is applicable also for other fermentation system. The basic assumption in the model is that the biomass can be divided into two parts (compartments)-an active part and a mainly inactive structural part. The size of the active part has a pivotal role in the model.
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  • 253
    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.
    Additional Material: 8 Ill.
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  • 254
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    Biotechnology and Bioengineering 38 (1991), S. 90-98 
    ISSN: 0006-3592
    Keywords: lactic acid bacteria ; mixed cultures ; population ratio ; pH ; temperature ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of pH, temperature, and inoculum composition on Streptococcus thermophilus 404 and Lactobacillus bulgaricus 398 mixed cultures were studied. Linear quadratic models according to these three variables were proposed to express the growth and acidification characteristics, as well as the final proportion between the two strains. Initial and operating conditions, allowing maximization of these characteristics were determined. Finally, a graphic method to predict both final total concentration and final proportion of the two bacteria as a function of pH, temperature and inoculum composition is presented.
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  • 255
    ISSN: 0006-3592
    Keywords: protease ; acyl transfer ; nucleophile efficiency ; inverse substrates ; trypsin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Benzyloxycarbonyl-L-alanine p-guanidinophenyl ester behaves as a trypsin “inverse substrate,” i.e., a cationic center is included in the leaving group instead of being in the acyl moiety. Using this substrate as an acyl donor, trypsin catalyzes the synthesis of peptide bonds that cannot be split by this enzyme. An optimal acyl transfer efficiency was achieved between pH 8 and 9 at 30°C.The addition of as much as 50% cosolvent was shown to be of minor influence on the acyl transfer efficiency, whereas the reaction velocity decreases by more than one order of magnitude. The efficiency of H-Leu-NH2 and H-Val-NH2 in deacylation is almost the same for “inverse” and normal type substrates.
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  • 256
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    Biotechnology and Bioengineering 38 (1991), S. 135-138 
    ISSN: 0006-3592
    Keywords: enzymic hydrolysis ; enzyme recovery ; process models ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: At the end of an enzymic hydrolysis process involving a solid lignocellulosic substrate, enzymes are found both in solution and absorbed to the substrate residue. Removal of residue from the system will result in loss of some of the enzymes, the extent of which will depend on the design of the process. To minimize enzyme loss, a study has been conducted in which six process models have been formulated and an enzyme loss function derived for each model based on the total amount of enzymes lost through residue removal. Model 1 is a reference model, characterized by an uninterrupted hydrolysis throughout the entire hydrolysis period. The residue is then washed in order to recover both sugar and adsorbed enzymes before the residue is discarded. Models 2-6 are all characterized by the removal of hydrolysate three times during the process, recirculation of dissolved and adsorbed enzymes to various points in the process and selection of a stage at which the residue is removed. The following conclusions could be drawn from the derived enzyme loss functions: Increased enzyme adsorption leads to increased enzyme loss.The enzyme loss decreases if the solid residue is removed late in the process.Both adsorbed and dissolved enzymes should be introduced at the starting point of the process. This is particularly important for dissolved enzymes. Three models were chosen for experimental studies, which are reported in a second, accompanying article. The experimental results obtained are compared with the theoretical study reported here.
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  • 257
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    Biotechnology and Bioengineering 38 (1991), S. 181-190 
    ISSN: 0006-3592
    Keywords: solventogenic metabolism ; NADH content ; NADH fluorescence ; continuous fermentation ; butanol biosynthesis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: With a constant glucose feed concentration, the change in the continuous culture dillution rate resulted in an altered fermentation profile and the cellular NADH content. The cultures growing at high dillution rates demonstrated an oxidative metabolism low NADH and butanol concentrations. The low specific NADH flourescence (F/X) at high butanol production rates suggested that a rapid regeneration of NADH to NAD is essential for a high solventogenic culture activity. The culture florescence and butanol concentration remained constant in the solventogenic dilution rate range of D = 0.05-0.2 h-1 with an inverse relationship between the specific flourescence (F/X) and the specific butanol production rate, qB. Flourometric NADH observations were confirmed by enzymatic NADH determination. The stiochiometric “Fermentation Equation” was used to check the experimental data consistency and to investigate the role of the available biosynthetic and reduction energy on the culture metabolic activities under different growth conditions. The butanol concentration in the broth was stabilized in a fed-batch process when the culture NADH fluorescence was being controlled through the addition of fresh medium.
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  • 258
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    Biotechnology and Bioengineering 38 (1991) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 259
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    Biotechnology and Bioengineering 38 (1991), S. 224-231 
    ISSN: 0006-3592
    Keywords: modeling ; immobilized ; growth ; Nitrobacter ; sensitivity analysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The modeling of the growth of Nitrobacter agilis cell immobilized in κ-carrageenan is presented. A detailed description is given of the modeling of internal diffusion and growth of cells in the support matrix in addition to external mass transfer resistance. The model predicts the substrate and biomass profiles in the support as well as the macroscopic oxygen consumption rate of the immobilized biocatalyst in time. The model is tested by experiments with continuously operated airlift loop reactors containing cells immobilized in κ-carrageenan. The model describes experimental data very well. It is clearly shown that external mass transfer may not be neglected. Furthermore, a sensitivity analysis of the parameters at their values during the experiments revealed that apart from the radius of the spheres and the substrate bulk concentration, the external mass transfer resistance coefficient is the most sensitive parameter for our case.
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  • 260
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    Biotechnology and Bioengineering 38 (1991), S. 254-259 
    ISSN: 0006-3592
    Keywords: light limitation ; shading ; maintenance ; mathematical model ; continuous culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Light-limited growth in continuous cultures of phototrophic organisms is modeled. It is assumed that light energy up-take rate depends hyperbolically on light intensity and that the maintenance costs are proportional to biomass. Modeling the light distribution caused by shading within the vessel is necessary to explain the existence of steady state in light-limited chemostats. The model fits well to experimental data from literature on light-limited chemostats and turbidostats. Attention is given to the implications of the model for the estimation of the specific maintenance rate constant in light-limited continuous cultures.
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  • 261
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    Biotechnology and Bioengineering 38 (1991), S. 304-313 
    ISSN: 0006-3592
    Keywords: Zymomonas mobilis ; molasses ; fermentation ; ethanol ; osmolality ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new osmotolerant mutant strain of Zymomonas mobilis was successfully used for ethanol production from beet molasses. Addition of magnesium sulfate to hydrolyzed molasses allowed repeated growth without the need of yeast extract addition. The kinetics and yields parameters of fermentation on media with different molasses concentrations were calculated. The anabolic parameters (specific growth rate, μ, and biomass yield, YX/S) were inhibited at elevated molasses concentrations while the catabolic parameters (specific ethanol productivity, qp, and ethanol yield, Yp/s) were not significantly affected. In addition to ethanol and substrate inhibition, osmotic pressure effects can explain the observed results.
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  • 262
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    Biotechnology and Bioengineering 38 (1991), S. 319-321 
    ISSN: 0006-3592
    Keywords: Protease ; acyl transfer ; nucleophile efficiency ; inverse substrates ; trypsin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Benzyloxycarbonyl-L-proline p-guanidinophenyl ester is an “inverse substrate” for trypsin; i.e., the cationic center is included in the leaving group instead of being in the acyl moiety. This substrate can be used in trypsin-catalyzed acyl-transfer reactions leading to the synthesis of Pro-Xaa peptide bonds. The reaction proceeds about 20 times slower than reaction with similar alanine-containing substrates, but the ratio between synthesis and hydrolysis is more favorable. The investigation of a series of nucleophiles led to information about the specificity of the process. Nucleophiles differing only in the P1′-position show an increasing acyl transfer efficiency in the order Phe 〈 Gly 〈 Ley 〈 Ser 〈 Ala 〈 lle. C terminal elongation of the nucleophiles is of minor influence on their efficiency. The formation of an H bond between the acyl-enzyme and the nucleophile seems to play an important role in the aminolysis of the acyl-enzyme.
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  • 263
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    Biotechnology and Bioengineering 38 (1991), S. 353-362 
    ISSN: 0006-3592
    Keywords: Rhizopus oligosporus ; fermentation ; starch ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A Semimechanistic mathematical model is developed which describes the growth of Rhizopus oligosporus in a model solid-state fermentation system. Equations are presented for the release of glucoamylase, the diffusion of glucoamylase, the hydrolysis of starch, the generation and diffusion of glucose, and the uptake of glucose and conversion into new biomass. Good agreement of the model with the experimental data was obtained only after the glucoamylase diffusivity and the maximum specific glucose uptake rate were altered from their originally determined values. The model recognizes the distributed nature of the solid-state fermentation and therefore is able to predict the concentration profiles of the system components within the substrate. The model provides an insight into the possible rate-limiting steps in solid-state fermentation - the generation of glucose within the substrate and the resulting availability of glucose at the surface.
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  • 264
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    Biotechnology and Bioengineering 38 (1991), S. 389-396 
    ISSN: 0006-3592
    Keywords: length-projected area ; length-number scaling ; Zoogloea ramigera ; Saccharomyces cerevisae ; fractal geometry ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The fractal nature microbial aggregates is a function of the type of microorganism and mixing conditions used to develop aggregates. We determined fractal dimensions from length-projected area (D2) and length-number scaling (D3) relationships. Aggregates of Zoogloea ramigera developed in rotating test tubes were both surface and mass fractals, with fractal dimensions of D2 = 1.69 ± 0.11 and D3= 1.79 ± 0.28 (±standard deviation), respectively. When we grew this bacteria in a bench-top fermentor, aggregates maintained their surface fractal characteristics (D2 = 1.78 ± 0.11) but lost their mass fractal characteristics (D3 = 2.99 ± 0.36). Yeast aggregates (Saccharomyces cerevisae) grown in rotating tests tubes had higher average fractal dimensions than bacterial aggregates grown under physically identical conditions, and were also considered fractal (D2 = 1.92 ± 0.08 and D3 = 2.66 ± 0.34). Aggregates porosity can be expressed in term of a fractal dimensions, but average porosities are higher than expected. The porosities of yeast aggregates (0.9250-0.9966) were similar to porosities of bacterial aggregates (0.9250-0.9966) cultured under the same physical conditions, although bacterial aggregates developed in the reactor had higher average porosities (0.9857-0.9980). These results suggest that that scaling relationships based on fractal geometry may be more useful than equations derived from Euclidean geometry for quantifying the effects of different fluid mechanical environments on aggregates morphology and characteristics such as density, porosity, and projected surface area.
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  • 265
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    Biotechnology and Bioengineering 38 (1991), S. 459-470 
    ISSN: 0006-3592
    Keywords: cell culture ; contact inhibition phenomena ; discrete mathematical model ; cell proliferation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We report the development of new class of discrete models that can accurately describe the contact-inhibited proliferation of anchorage-dependent cells. The models are based on cellular automata, and they quantitatively account for contact inhibition phenomena occurring during all stages of the proliferation process: (a) the initial stage of “exponential” growth of cells without contact inhibition; (b) the second stage where cell colonies form and grow with few colony mergings; and (c) the final stage where proliferation rates are dominated by colony merging events. Model prediction are presented and analyzed to study the complicated dynamics of large cell populations and determine how the initial spatial cell distribution, the seeding density, and the geometry of the growth surface affect the observed proliferation rates. Finally, we present a model variant that can simulate contact-inhibited proliferation of asynchronous cell populations with arbitrary cell cycle-time distribution. The latter model can also compute the percentage of cells that are in a specific phase of their division cycle at a given time.
    Additional Material: 17 Ill.
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  • 266
    ISSN: 0006-3592
    Keywords: energy balances ; Saccharomyces cerevisiae ; fermentation ; microcalorimetric monitoring ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Energy balance calculations were performed for different physiological states during batch growth of Saccharomyces cerevisiae with glucose as carbon and energy source. For the different physiological states, energy recoveries close to one were obtained, which permitted a continuous control that the constantly changing growth process was quantified accurately. During the respiro-fermantative phase of growth, during which glucose served as the carbon and energy source, a low-heat-yield value (ΔQx) of -8.6 kJ/g dry biomass formed was obtained. This low-heat-yield value was due to the mainly fermentative metabolism during the middle of this phase of growth. After a transition phase, the ethanol produced during the respiro-fermentative growth was respired. During this respiratory phase, the heat yield values increased markedly, resulting in a lowest value of -42.7 kJ/g. The low-heat-yield values of the respiro-fermentative growth is not a reflection of the most efficient metabolism of S. cerevisiae. On the contrary, during the middle of this phase, 74% of the energy input was dissipated as ethanol, 6% was dissipated as heat, and the energy conserved as biomass was just 13%, while during the early respiratory phase, 69% of the energy input was dissipated as heat, and 22% of the energy input was conserved as biomass. By mathematical modeling and direct monitoring on-line of the rate of heat production, continuous calculations of (1) glucose consumption, and (3) biomass production were performed, and were shown to correlate closely with measured values for the continuously changing growth process.
    Additional Material: 8 Ill.
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  • 267
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    Biotechnology and Bioengineering 38 (1991), S. 499-506 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Bed fluidization offers the possibility of gaining the advantages of fixed-film biological processes without the disadvantage of pore clogging. However, the biofilm detachment rate, due to hydrodynamics and particle-to-particle attrition, is very poorly understood for fluidized-bed biofilm processes. In this work, a two-phase fluidized-bed biofilm was operated under a constant surface loading (0.09 mg total organic carbon/cm2 day) and with a range of bed height (H), fluid velocities (U), and support-particle concentrations (Cp). Direct measurements were made for the specific biofilm loss rate coefficient (bs)and the total biofilm accumulation (XfLf). A hydrodynamic model allowed independent determination of the biofilm density (Xf), biofilm thickness (Lf), liquid shear stress (τ), and Reynolds number (Re). Multiple regression analysis of the results showed that increased particle-to-particle attrition, proportional to Cp and increased turbulence, described by Re, caused the biofilms to be denser and thinner. The specific detachment rate coefficient (bs) increased as Cp and Re increased. Almost all of the 6, values were larger than predicted by a previous model derived for smooth biofilms on a nonfluidized surface. Therefore, the turbulence and attrition of bed fluidization appear to be dominant detachment mechanisms.
    Additional Material: 6 Ill.
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  • 268
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    Biotechnology and Bioengineering 38 (1991), S. 545-551 
    ISSN: 0006-3592
    Keywords: rice ; α-amylase ; callus ; mRNA ; and 2,4-dichlorophenoxyacetic acid (2,4-D) ; plant cell fermentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Rice seed callus expressed and secreted α-amylase at high levels. Twenty percent of the protein secreted by the callus was αamylase. The callus secreted about 840 μg α-amylase with 10.9 × 103 units of activity per gram dry weight callus per day. The α-amylase from callus exhibited a more complex isoform pattern than the germinating seed α-amylase. In addition, the level of mRNA expression by the five α-amylase gene groups was markedly different between callus and the germinating seed. The rice callus culture has features which it attractive as a potential system for expression proteins in plant cell fermentation systems.
    Additional Material: 3 Ill.
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  • 269
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    Biotechnology and Bioengineering 38 (1991), S. 557-560 
    ISSN: 0006-3592
    Keywords: levan ; continuous culture ; molecular weight ; Erwinia herbicola ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The optimal production of the fructan biopolymer levan by the bacterium Erwinia herbicola was investigated, including variations in nitrogen, carbon and phosphorous sources, pH, incubation time, culture yields up to 19% by weight produced based on conversion of sucrose as the carbon source when grown in a continuous culture system and processed by tangential flow filtration. Product identity was confirmed with gas chromatography (GC) and 13C nuclear magnetic resonance (NMR). Gel permeation chromatography (GPC) and low-angle laser light scattering (LALLS) determination of the molecular weight of the product showed a significant difference in molecular weight values dependent on the method of analysis. Analysis by GPC resulted in molecular weight one order of magnitude lower than LALLS independent of sample, underscoring the unusual nature of this biopolymer.
    Additional Material: 2 Ill.
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  • 270
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    Biotechnology and Bioengineering 38 (1991), S. 588-602 
    ISSN: 0006-3592
    Keywords: endothelium ; genetic expression ; protein synthesis ; shear stress ; signal transduction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Mammalian cells responds to physical forces by altering their growth rate, morphology, metabolism, and genetic expression. We have studied the mechanism by which these cells detect the presence of mechanical stress and convert this force into intracellular signals. As our model systems, we have studied cultured human endothelial cells, which line the blood vessels and forms the interface between the blood and the vessel wall. These cell responds within minutes to the initiation of flow by increasing their arachidonic acid metabolism and increasing the level of the intracellular second messengers inositol trisphosphate and calcium ion concentration. With continued exposure to arterial levels of wall shear stress for up to 24 h, endothelial cells increase the expression of tissue plasminogen activator (tPA) and tPA messenger RNA (mRNA) and decrease the expression of endothelin peptide and endothelin mRNA. Since the initiation of flow also causes enhanced convective mass transfer to the endothelial cell monolayer, we have investigated the role of enhanced convection of adenosine trisphosphate (ATP) to the cell surface in eliciting a cellular response by monitoring cytosolic calcium concentrations on the single cell level and by computing the concentration profile of ATP in a parallel-plate flow geometry. Our result demonstrate that endothelial cells respond in very specific ways to the initiation of flow and that mass transfer and fluid shear stress can both play a role in the modulation of intracellular signal transduction and metabolism.
    Additional Material: 9 Ill.
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  • 271
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    Biotechnology and Bioengineering 38 (1991), S. 637-652 
    ISSN: 0006-3592
    Keywords: metabolites ; siderophores ; iron transport ; metabolic regulation ; cybernetic modeling ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The cybernetic modeling framework developed by Ramkrishna and co-workers has been applied to a case of bacterial metabolite production, namely the production of siderophores (iron-chelating agents) associated with iron-limiting fermentation conditions. Experimental growth data showed that, even though final biomass levels were controlled by exhaustion of the carbon source, iron-limiting conditions also affected the biomass yield. A structured model which includes the process of an iron-limiting energy resource production was able to quantitatively account for this apparent dual-substrate limitation over a wide range of batch and continuous operating conditions. The experiments data also showed quite large difference in iron uptake over the wide range of operating condition and iron levels investigated. The inclusion in the model of the processes of low and high (siderophore-mediated) affinity iron transport, and siderophore production led to simulation results that were in good quantitative agreement with the siderophore, medium and cell iron levels, in both batch and steady-state continuous culture operating conditions.
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  • 272
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    Biotechnology and Bioengineering 38 (1991), S. 665-677 
    ISSN: 0006-3592
    Keywords: hybridoma ; flow cytometry ; cell cycle ; population balance ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Single-cell rates of accumulation of cellular protein have been determined as a function of total protein content using flow cytometry and population balance equations for exponentially growing murine hybridoma cells in the individual G1, S1 and G2 + M cell cycle phases. A novel flow cytometric technique for the identification of hybridoma cells in mitosis was developed and implemented. The data were obtained from a producer cell line which synthesizes and secretes high levels of monoclonal antibodies, and from a nonproducer clone which does not synthesize and secrete substantial amounts of antibody. The results indicate that the kinetics of single-cell protein accumulation in these two cell lines are considerably different. In particular, low protein content G1 phase producer cells were characterized by a rate of protein accumulation which was approximately five times higher than the mean rate observed for higher protein content producer cells cycle phase. In contrast, the rate of accumulation of protein increased continuously with totalprotein content for the G1 phase nonproducer cells. S phase hybridoma cells were characterized by a considerably lower rate of protein accumulation which did not vary much with protein content for either cell line. Finally, G2 + M phase producer cells demonstrated a negative rate of protein accumulation which indicates that the rates of protein synthesis. It was hypothesized that these differences in total protein accumulation are caused by differences in monoclonal antibody accumulation. The distribution of rates suggests the need for a segregated approach to the modeling of the kinetics of antibody production in hybridomas.
    Additional Material: 10 Ill.
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  • 273
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    Biotechnology and Bioengineering 38 (1991), S. 719-726 
    ISSN: 0006-3592
    Keywords: microsin B17 promoter ; fusion strain ; gene expression ; growth rate dependence ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Prior work has demonstrated that the microsin antibiotics are produced by enteric bacteria when the growth medium is depleted of nutrients. Because the control loci could have biotechnical potential, and general stress-response phenomena are of importance to understanding how bacteria survive in natural and bioreactor environments, we examined further the growth rate dependence of gene expression under the control of the microsin B17 promoter. This work entailed performing batch and chemostat growth experiments with a strain of E. coli K-12 containing a mcbA-lacZ gene fusion in the chromosome. Our results indicate that when a culture is presented with excess respiratory substrate, a well defined growth rate exists, below which a significant induction event occurs. However, cultures that are fermenting or highly glycolytic tend to express poorly. Additionally, the utility of the fusion strain was examined by performing fed-batch cultivation experiments. We found that sustained production in a fed-batch reactor can be accomplished by using a straightforward, exponential nutrient feeding profile.
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  • 274
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    Biotechnology and Bioengineering 38 (1991), S. 749-760 
    ISSN: 0006-3592
    Keywords: metabolic modeling ; bioreactor optimization ; recombinant protein synthesis ; induction dynamics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The dynamics of chemically induced chloramphenicolaceyl-transferase (CAT) expression are determined in batch cultures of Escherichia coli DH5αF'IQ [pKK262-1]. This article is directed towards understanding the coupling of induced cloned-protein synthesis and reduced cell growth which are balanced in the optimal system. Experimental results indicate that the best inducer (IPTG) concentration is near 1.0 mM when added during midexponential growth. Lower concentrations cause only weak induction, whereas higher concentrations cause sufficiently strong induction that cell growth is suppressed. Induction at the onset of the stationary phase results in high expression but is accompanied by stimulated protease activity. Also, cell mass yield is adversely affected by enhanced protein synthesis. A structured metabolic model is shown to predict the responses of instantaneous growth rate and productivity which result from protein overexpression. This model can be employed to predict alternative reactor strategies and operating conditions necessary for the design of efficient bioprocess.
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  • 275
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    Biotechnology and Bioengineering 38 (1991), S. 802-802 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 4 Ill.
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  • 276
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    Biotechnology and Bioengineering 38 (1991), S. 803-804 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 277
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    Biotechnology and Bioengineering 38 (1991), S. 838-852 
    ISSN: 0006-3592
    Keywords: hybridoma cells ; cell cycle ; morphology ; geometry ; mechanical properties ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Morphological, geometrical, and rheological properties of the GAP A3 hybridoma cell line have been evaluated as a function of the cell cycle. Interference contrast video microscopy and scanning electron microscopy (SEM) showed that a sample of cells taken from the middle of the exponential growht phase displayed a range of cell morphologies, consistent with a heterogeneous growing culture. Micropipet manipulation was used to measure the geometrical (cell volume) and mechanical (cortical tension and apparent cell viscosity) properties of single cells selected at random from a sample in the middle of the exponential growth phase. Consistent with the range of morphologies, cell volumes (1400 to 5700 μm3) and apparent viscosities (430 to 1.2 × 104 P) showed a wide range of values at 37°C, demonstrating that a hybridoma cell line cannot be characterized by a single value for any one property, and that properties must be related to their cycle dependence when considering proliferating cells. Direct, video-microscopic observation of synchronized cells, and of individual cells that were followed throughout their cell cycle, allowed us to correlated distinct morphologies with phases of the cell cycle. As the cell cycle progresses, an increase in cell volume by a factor of 3 to 4is accompanied by an overall increase in apparent cell viscosity by approximately the same ratio, consistent with an accumulation of more cytoplasmic material in the older cells. Also, a decrease in average apparent viscosity by a factor of 10. These results are important in order to evaluate the possible role of certain structural, cell-cycle dependent features in shear and abrasion sensitivity. This is a problem of current concern in the bioreactor culture of mammalian cells.
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  • 278
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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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  • 279
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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.
    Additional Material: 11 Ill.
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  • 280
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    Biotechnology and Bioengineering 38 (1991), S. 929-940 
    ISSN: 0006-3592
    Keywords: culture contamination ; photon correlation spectroscopy ; dynamic light scattering ; particle sizing ; mixed culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The application of photon correlation spectroscopy (PCS) to detect culture contamination in chemostats was studied. It was found that the presence of a given particle size in a population of particles of a different size could be detected, but this ability was strongly dependent on particles of a different size could be detected, but this ability was strongly dependent on particle size difference and was most sensitive when contaminants are larger than the host. The inherent polydisparity of actively growing and dividing microbial cells negates any advantage in the use of multi-angle PCS to detect contaminants.
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  • 281
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    Biotechnology and Bioengineering 38 (1991), S. 960-960 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 282
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    Biotechnology and Bioengineering 38 (1991), S. 972-976 
    ISSN: 0006-3592
    Keywords: cell culture ; antibody production ; fermentation ; continuous culture ; cell growth ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A clonal derivative of a transfectant of the SP2/O myeloma cell line producing a chimeric monoclonal antibody was maintained in steady-state, continuous culture at dilution rates ranging from 0.21 to 1.04 day-1. The steady-state values for nonviable and total cell concentrations increased as the dilution rate decreased, while the viable cell concentration was roughly independent of the dilution rate. At steady state, the specific growth rate increased and the specific death rate decreased as the dilution rate increased. The maximum specific growth rate was 1.15 day-1. Antibody production was growth associated and the specific rate of antibody production increased linearly as the specific growth rate increased.
    Additional Material: 6 Ill.
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  • 283
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    Biotechnology and Bioengineering 38 (1991), S. 995-1000 
    ISSN: 0006-3592
    Keywords: tubular-loop photobioreactor ; orientation ; Chlorella pyrenoidosa ; biomass productivity ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The profiles of photon flux density incidented on a tubularloop photobioreactor in the day could be altered by inclining the bioreactor at an angle with the horizontal. The photon flux density at noon decreased with increasing angle of inclination, whereas the photon flux density in the early morning and late afternoon increased with increasing angle of inclination. The overall photosynthetic radiance received by the bioreactor inclined at 0, 25, 45, and 80° was 1:0.89:0.77:0.62. Regardless of the angle of bioreactor inclination, the overall biomass output rate of a fed-batch culture over an 8-h/day period was comparable (26-36 g-biomass m-2 bioreactor surface area day-1). As a bioreactor inclined at an angle occupied smaller land area, and daily biomass output rate per land area of a bioreactor inclined at 80° (130 g-biomass m-2 land) was about six times of that obtainable at horizontal position (21-g biomass m-2 land). The bioenergetics growth yield from the absorbed photosynthetic radiance was not a constant but an inverse function of the photon flux density. The quasi-steady state chlorophyll content of the Chlorella cells varied between 36 and 63 mg g-1 cells. Photoinhibition of the maximum photosynthetic capacity was not observed in this study.
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  • 284
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    Biotechnology and Bioengineering 38 (1991), S. 1029-1033 
    ISSN: 0006-3592
    Keywords: Aspergillus niger ; 3,4-dihydroxyphenylacetaldehyde ; dopamine ; monoamine oxidase ; norlaudanosoline ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Norlaundanosoline is a key intermediate in the synthesis of the benzylisoquinoline alkaloids providing the upper isoquinoline portion of the morphinan skeleton. This study evaluates the feasibility of using Aspergillus niger as an in situ biotransformation system to produce norlaudanosoline from dopamine. A. niger was chosen because monoamine oxidase can be readily induced in this organism. Monoamine oxidase catalyzes the conversion of dopamine to 3,4-dihydroxyphenylacetaldehyde. In the presence of dopamine, this aldehyde will then undergo a spontaneous Picket-Spengler condensation to form norlaudanosoline. Fermentation condition to form norlaudanosoline. Fermentation conditions were optimized for the production monoamine oxidase by using a two-stage process consisting of a growth stage and an induction stage. pH control was found to be important, and at pH 4.5 dopamine accumulation in the cells was high as was the level of monoamine oxidase. With pH control at 4.5, up to 21% of the cellular dopamine was converted to norlaudanosoline. It is proposed that with further protein engineering improvements, this system may prove suitable for the in situ bio-transformation of dopamine to norlaudanosoline.
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  • 285
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    Biotechnology and Bioengineering 38 (1991), S. 1020-1028 
    ISSN: 0006-3592
    Keywords: hybridoma ; cell culture ; continuous culture ; kinetics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A hybridoma cell line, AFP-27-P, was cultivated in continuous culture under glucose-limited conditions. The viable cell concentration, dead-cell concentration, and cell volume all varied with the dilution rate. A model previously developed for a nonproducing clone of the same cell line, AFP-27-NP, was extended to describe the behavior of the cells. The relationship between the specific growth rate and glucose concentration is described by a function similar to the Monod model. A threshold glucose concentration and a minimum specific growth rate are incorporated; the model is meaningful only at glucose concentration and a minimum specific growth rate are incorporated; the model is meaningful only at glucose concentrations and specific growth rates above these levels. The relationship between the death rate and the glucose concentration is described by an inverted Monod-type function. Furthermore, the yield coefficient based on glucose is constant in the lower range of specific growth rates and changes to a new constant value in the upper range of specific growth rates. No maintenance term for glucose consumption is used; in the plot of specific glucose consumption rate vs. specific growth rate, the line intercepts the specific growth rate at a value close to the minimum growth rate. The productivity of antibody as a function of the specific growth rate is described by a mixed type model with a noon-growth-associated term and a negative-growth-associated term. The values for the model parameters were determined from regression analysis of the steady state data.
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  • 286
    ISSN: 0006-3592
    Keywords: catabolite repression ; protein A ; membrane proteins ; continuous culture ; protein expression ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Although widely used in experimental and industrial situations, genetically engineered plasmids containing the lac promoter from Escherichia coli are subject to catabolite repression when grown in glucose-containing media. Several methods of overcoming this problem have been investigated by studying the expression of the protein A gene from Staphylococcus aureus under the control of the Escherichia coli lac promoter. When glycerol is used as a sole carbon source, the plasmid is unstable and is rapidly lost from the culture. When the bacteria are grown in chemostats under glucose limitation, the plasmid is maintained, even at high dilution rates, and the expression of protein A is similar to that observed when glycerol was used. The balance between metabolic load and protein A expression seems to be maintained by reducing the gene dose to a tolerable level. Depending on the metabolic conditions prevailing in the culture, this is achieved, either by reducing the copy number of the plasmid or in extreme cases by removing the plasmid altogether.
    Additional Material: 10 Ill.
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  • 287
    ISSN: 0006-3592
    Keywords: chloramphenicol acetyl transferase ; baculovirus ; Spodoptera frugiperda ; serum-free medium ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Spodoptera frugiperda insect cells were grown in Sf-900 serum-free medium and two kinds of serum-supplemented media (IPL -41 and Grace's). The specific growth rates of uninfected cells were found to be 0.024, 0.35, and 0.034 h-1 respectively, at 33°C. The IPL -41 medium supported to highest maximum cell density (10.6 × 106 cells/mL) compared to 3.5 × 106 and 8.7 × 106 cells/mL with the Grace's and serum-free media, respectively. In temperature shifdown experiments with a temperature-sensitive baculo-virus (acts10YM1CAT), virus titer and chloramphenicol acetyl transferase (CAT) expression were highest in the IPL -41 (5.1 × 107 PFU/mL and 20000 U/mL). Use of Grace's medium gave higher virus titers than the serum-free medium (4.4 × 106 vs 4.1 × 105 PFU/mL) as well as higher CAT titers (7050 vs 1980 U/mL). Interestingly, in the three media used, the highest virus and CAT titers were obtained at MOI (multiplicity of infection) of 0.02 At MOI of 2.0 virtually no increase in virus of CAT titer was observed. This result is contrary to those obtained at constant-temperature (27°C) infection and cell culture, in which higher virus titers and recombinant protein expression and obtained at higher MOI.
    Additional Material: 5 Ill.
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  • 288
    ISSN: 0006-3592
    Keywords: plant cell suspensions ; carbon utilization ; growth yield ; maintenance coefficient ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Methodology is presented for the determination of growth yield (Yg) and maintenance coefficient (m) for carbon utilization of plant cells grown in suspension culture. Estimation of Yg and m requires measurements of specific growth rate (µ) and specific rate of substrate uptake (q) at different growth limiting substrate concentrations. Batch culture of tobacco cells did not permit evaluation of Yg and m because µ is constant and maximal during most of the growth cycle. In batch culture, the period of declining specific growth rate is extremely brief because of the rapid transition from logarithmic growth to stationary phase. This occurs because the Km for growth is relatively small compared to the initial sucrose concentration. Thus, when the substrate level reaches the Km, the large mass of cells rapidly depletes the remaining substrate. In contrast, semicontinuous culture facilitates the determination of Yg and m because various steady-state growth rates can be achieved. Mathematical expressions were developed to determine the effective values of µ and q over the semicontinuous replacement interval. The validity of this approach was verified by conducting simulations using experimentally determined parameters.
    Additional Material: 5 Ill.
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  • 289
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    Biotechnology and Bioengineering 38 (1991), S. 1159-1165 
    ISSN: 0006-3592
    Keywords: continuous glycerolysis ; lipase adsorbed on liposome ; microemulsion ; reversed micelles ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Chromobacterium viscosum lipase which has adsorbed on liposome and solubilized in microemulsion droplets of glycerol containing a little amount of water could catalyze the glycerolysis of olive oil. Studies on the continuous glycerolysis of olive oil by the immobilized enzyme was done at 37°C in continuous stirred vessel bioreactor with polysulfone membrane. The effect of the flow rate of substrate (olive oil) in isooctane on the conversion and composition of the outlet was investigated using high-performance liquid chromatography (HPLC). The conversion increased with decrease in the flow rate. And we studied the effect of water content in the glycerol-water-lipase solution on the glycerolysis reaction. The conversion to desirable products, mono- and di-olein, was improved without a substantial production of oleic acid at lower water concentrations, i.e., below 8.0% (w/v) which corresponds to a wo value of 0.97. At water concentration higher than 8.0% (w/v), the amount of free fatty acid was dramatically increased. Higher operational stability of the enzyme reactor, and the half-line of the enzyme continuous reaction was about 7 weeks.
    Additional Material: 9 Ill.
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  • 290
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 1190-1202 
    ISSN: 0006-3592
    Keywords: continuous centrifugal bioreactor ; ethanol fermentation ; mammalian cell hybridoma ; monoclonal antibody ; fluidized bed ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A continuous centrifugal bioreactor (CCBR), developed to study the growth and productivity of dense suspensions cultures, has been applied to both fermentation and mammalian cell cultivation processes. With this approach, high-density nonflocculent cultures are maintained in a tapered fluidized bed by balancing the drag forces on the cells due to following substrate with the centrifugal forces. The Sysyem was first used to produce ethanol by fermentation with Saccharomyces cerevisiae; then with H21A1 mouse hybridoma cells secreting monoclonal antibody (MoAb), lgM. Results of this research show the feasibility of using the CCBR for both production of secreted products and as a research tool for studying cell metabolism and production kinetics. Media recycle may be used to modify the behavior of the system form a plug flow apparatus to a continuous stirred reactor (CSTR).
    Additional Material: 8 Ill.
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  • 291
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 1218-1222 
    ISSN: 0006-3592
    Keywords: horse liver alcohol dehydrogenase ; lipase ; optical resolution ; organometallic compounds ; second harmonic generation ; organometallic compounds ; second harmonic generation ; nonlinear optical material ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Twelve species of optically active metallocene derivatives having a 4-nitrophenyl group were prepared with horse liver alcohol dehydrogenase- or lipase-catalyzed optical resolution as the key step. The second harmonic generation (SHG) efficiently of these products was measured by the power method using the fundamental light from the Nd:YAG laser. (-)-1- (4-Nitrophenylthio)ethylferrocene (9), (+)-1-(4-nitrophenylthio)ethylruthenocene (16), (+)-1-(4-nitrophenylthio)ethylosmocene (19), (+) -1-(5-nitro-2-pyridylthio)ethylruthenocene (21), and (+) -1-[(4-nitrophenylhydrazono)methyl] -2-methylferrocene (12) showed SHG signals. The highest SHG efficiency was found with (+) -16, being 27 times more intense than the commonly used urea standard.
    Additional Material: 2 Ill.
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  • 292
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 1253-1258 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 3 Ill.
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  • 293
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    Biotechnology and Bioengineering 38 (1991), S. 1280-1284 
    ISSN: 0006-3592
    Keywords: Saccharomyces cerevisiae ; biotransformation ; oxidoreductases ; carbonyl ; stereospecific ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The progress of reductive biotransformations of a variety of earbonyl compounds by whole cells of baker's yeast was monitored with time. Biotransformations rates ranged from 0.11 to 112.12 mg product formed per g dry yeast per h. While rapid biotransformations of citronellal and ethyl benzoylformate were observed, complete conversion of substrate to product did not occur. Reductive conversions of ethyl- and methyl-acetoacetate went to completion in 6 and 12 h respectively. Ethyl mandelate was produced stereoselectively, favoring the (R)- stereoisomer and ethyl and methyl-3-hydroxybutyrate were produced with (S)-enantiospecificity. Yeast crude extract and resuspended presence of NAD(P)H. Ethyl benzoylformate and methyl-and ethyl-acetoacetate were preferentially reduced by yeast crude extract as compared to resuspended pellet and, in the case of the former two substrates, the reaction manifested a preference for NADPH over NADH.
    Additional Material: 8 Ill.
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  • 294
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    Biotechnology and Bioengineering 38 (1991), S. 1302-1307 
    ISSN: 0006-3592
    Keywords: liquid-liquid extraction ; selective separation of proteins ; reversed micelles ; purification ; lipases ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Selective separation and purification of two lipases form Chromobacterium viscosum were carried out by liquid-liquid extraction using a reversed micellar system. Optimum parameters for extraction were determined using a 250 mM AOT micellar solution in isooctane. Complete separation of the two lipases was achieved at pH 6.0 with a 50mM potassium phosphate buffer solution containing 50 mM KCI. By adding 2.5% by volume of ethanol to the lipase-loaded micellar solution, 85% of the extracted lipase could be recovered in a new aqueous phase, 50 mM K2HPO4 with 50 mM KCl, at pH 9.0. Lipase A was purified 2.6-fold with a recovery of 86%, and lipase B by 1.5-fold with a recovery of 76%.
    Additional Material: 9 Ill.
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  • 295
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 1331-1336 
    ISSN: 0006-3592
    Keywords: plasmid ; yeast ; detection ; sensor ; image ; analysis ; 5-fluoro-orotic acid ; determination ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel imaging sensor system for the determination of plasmid carrying yeast cells was developed. The sensor system consisted of an Silicon Intensifier Target (SIT) video camera, a fluorescent microscope, and a personal computer system equipped with an image memory board. This system was based on the fact that the membrane integrity of only plasmid-carrying cells is lost following cell growth in 5-fluoro-orotic acid (5-FOA) containing medium, and consequently these target cell can be stained with fluorescent probes and detected. In this study, plasmid-carrying cells were detected and their fraction determined in a mixture of both plasmid-carring and plasmid-free cells. A good correlation was observed between the values determined by this sensor system and the conventional method in the 30%-80% range, and one assay was possible within 4 h. This sensor system could be used for the monitoring of plasmid-carrying fraction in recombinant yeast cells during cultivation.
    Additional Material: 6 Ill.
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  • 296
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    Biotechnology and Bioengineering 37 (1991), S. 557-566 
    ISSN: 0006-3592
    Keywords: anaerobic digestion ; fluidized-bed ; biofilm formation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of various operating variables such as initial inoculum circulation, dilution rate, chemical oxygen demand (COD) loading rate, and quantity and quality of inoculum on the process of film formation on sand surface and reactor performance were studied using synthetic glucose based wastewater. It was found that the film formation process is favored by a high dilution rate, a large quantity of inoculum, and an inoculum having high methane producing capacity. Experimental observations indicate that the biofilm formation process is initiated by methanogenic bacteria.
    Additional Material: 15 Ill.
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  • 297
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 298
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    Biotechnology and Bioengineering 37 (1991), S. 597-607 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new adaptive DO (dissolved oxygen) concentration control algorithm considering DO electrode dynamics with response time delay has been developed. A system model with two time-varying parameters was used to relate the DO concentration with two control variables: air flow rate and agitation speed. Parameters of this model were estimated on-line using a regularized constant trace recursive least-squares method. An extended Kalman filter was used to remove the effect of noises from the DO concentration measurements and thus to improve control performance. A discrete one-step ahead control scheme was adopted to determine control actions based on the parameter estimation results. Experimental results showed that the new adaptive DO concentration control algorithm performed better than other algorithms tested, a PID controller and adaptive algorithms without the DO electrode dynamics.
    Additional Material: 9 Ill.
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  • 299
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991), S. 639-646 
    ISSN: 0006-3592
    Keywords: enzymatic synthesis ; anhydrous pyridine ; trifluoroethylbutyrate ; transesterification ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A variety of enzymes have been found to acylate sucrose in anhydrous pyridine. The enzymic reaction is highly selective; with trifluoroethylbutyrate as ester donor, enzyme-catalyzed transesterification of sucrose yielded sucrose 1′-butyrate and sucrose 6, 1′-dibutyrate. No sucrose-tributyrates were formed. Using a similar technique, a long-chain linear sucrose polyester has been prepared using Proleather, an alkaline protease from a Bacillus sp. This protease catalyzes the esterification of sucrose with bis(2, 2, 2-trifluoroethyladipate) in a 1:1 ratio to yield a sucrose-containing polyester with Mw = 2100 and Mn = 1600 for a polydispersity of 1. 31. Polymers with molecular weights in excess of 13, 000 have been prepared by this enzymic approach, indicating that molecules containing over 30 sucrose units have been produced. The polyester is extremely water soluble and soluble in polar organic solvents. As with the sucrose dibutyrate, the polyester has ester linkages at the C6 and C1′ positions on the sucrose. The polyester can be depolymerized using Proleather in aqueous buffer, pH7. After 9 days in aqueous buffer, Proleather catalyzed the breakdown of the polyester to an Mw of ca. 900. This sucrose-containing polyester may have applications as a water-absorbent, biodegradable plastic for use as diapers and hygienic products, water-treatment chemicals, and components of drug delivery systems.
    Additional Material: 5 Ill.
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  • 300
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    Biotechnology and Bioengineering 37 (1991), S. 97-102 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Solvent selection tests were carried out for the Δ1 dehydrogenation of 6-α-methylhydrocortisone-21-acetate by Arthrobacter simplex cells in the presence of organic solvents. Solubility limits were determined for substrate and product in dry and water-saturated solvents and solvent mixtures. Molecular toxicity levels were estimated by measuring the dehydrogenation activity decay of freely suspended cells incubated in solvent-saturated aqueous media. Chloroform and n-decan-1-ol were the best choice of solvent, for both solubility and catalytic stability. High concentrations of water-soluble additives, such as monosodium glutamate, were found to greatly improve the retention of activity in chloroform-saturated media.
    Additional Material: 7 Ill.
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