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  • 1985-1989  (3,097)
  • 1830-1839
  • Biochemistry and Biotechnology  (3,097)
  • 201
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 34 (1989), S. 731-740 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The influence of Aspergillus niger broth rheology, bioreactor geometry, and superficial gas velocity on the volumetric liquid phase oxygen transfer coefficient (kLaL), riser gas holdup (εGR), and circulating liquid velocity (uLR) was studied in a bubble column (BC) and two external-circulation-loop airlift (ECLAL) bioreactors. The results are compared to those of previous studies on homogeneous fluids and in particular with a recent study on non-Newtonian carboxymethylcellulose (CMC) solutions conducted in the same contactors used for the A. niger fermentations. As expected from the CMC-based studies, in the heterogeneous broths of A. niger εGR, kLaL, and uLR decreased with increasing broth apparent viscosity; εGR and kLaL decreased with increasing downcomer-to-riser cross-sectional area ratio, Ad/Ar, whereas uLR increased with increasing Ad/Ar. Gas holdup data in the airlift fermentations of A. niger were well predicted by the CMC-based correlation. However, the CMC-based correlations produced conservative estimations of kLaL and overestimates of uLR compared to the observed values in the A. niger broths.
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  • 202
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 34 (1989), S. 785-793 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The fermentation of gaseous substrates such as CO, H2, and CO2 may be performed in a continuous stirred tank reactor, as well as the traditional batch reactor. In this article, the conversion of carbon monoxide by Peptostreptococcus productus is demonstrated in a stirred tank reactor under both mass transfer-controlled and nonmass transfer-controlled conditions. Utilizing a non-steady-state procedure, intrinsic rates are evaluated under non-mass transfer-controlled conditions in a time period of only 5-6 hours. A steady-state procedure was used to evaluate CSTR performance under mass transfer-controlled conditions. The mass transfer coefficient was calculated, followed by the development of a model to predict CSTR behavior for this gas phase substrate.
    Additional Material: 7 Ill.
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  • 203
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 34 (1989), S. 838-843 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In view of the biochemical reaction catalyzed by enzyme powder suspended in a water-insoluble organic solvent, an equation was derived to estimate the amount of water bound to the enzyme powder. With this equation, an apparent adsorption isotherm between free water (water freely dissolved in benzene) and bound water (water bound to crude lipase powder of Pseudomonas fluorescens) was obtained. A direct lactonization reaction (synthesis of cyclopentadenolide from 15-hydroxypen-tadecanoic acid) catalyzed by crude lipase powder of Pseudomonas fluorescens was carried out batchwise in microaqueous benzene at 40oC. A kinetic model of the enzymatic reversible lactonization reaction was derived, from which the effect of moisture content on the initial reaction rate with a fully hydrated enzyme was mathematically expressed. The observed initial reaction rate first increased, then decreased with increasing moisture content, giving rise to the maximum rate at a certain level of the moisture content. The drop in the reaction rate at lower moisture content was due to a lesser hydration of the enzyme molecule (hydration-limited) and the decrease in the reaction rate at higher moisture content was attributed to the dependence of the true initial rate of the reversible reaction on the moisture content (true reversible reaction limited), and could be simulated by the kinetic model. The equilibrium yield approached 100% at a lower moisture content.
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  • 204
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 34 (1989), S. 868-871 
    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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  • 205
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 34 (1989) 
    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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  • 206
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 34 (1989), S. 896-901 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Equilibrium adsorption studies on Escherichia coli and Saccharomyces sp. revealed the capacity and affinity of these organisms for the surfaces of powdered charcoal and nickel. In simple salt solutions both organisms readily adsorbed to each solid with an affinity and maximum loading capacity individual to each cell-solid combination. In the presence of common growth media (lab-lemco, nutrient broth, peptone, and yeast extract, individually at a concentration of 1.3%), each medium substantially inhibited adsorption. Each medium contained a protein-aceous constituent as determined by ultraviolet (UV) analysis. The degree of inhibition was relative to medium concentration present during assay. Cell wall extracts from whole-yeast cells also effectively inhibited adsorption. Cells adsorbed in the presence of sodium chloride solutions were susceptible to subsequent desorption by nutrient broth.
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  • 207
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 34 (1989), S. 1085-1091 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The desire to increase cell density and product concentration has been the primary driving force for the development of better animal cell culture processes. In the technique used in our laboratory - microencapsulation - insect cells (Spodoptera frugiperda), infected with a temperature-sensitive mutant of the Autographa californica nuclear polyhedrosis virus (AcNPV), were cultured in multiple membrane alginate-polylysine (PLL) microcapsules which had a controlled membrane molecular-weight cutoff and an intracapsular alginate concentration which was ca. 16% lower than that obtained in the commercially available single-membrane system. Cell culture experiments indicated that the intracapsular alginate concentration appears to be a key factor in achieving good cell growth. It was possible to obtain intracapsular cell densities of 8 × 107 cells/mL capsules and virus concentrations to 109 IFU/mL capsules. The virus litre in the supernatant was ca. 300 times lower, indicating that virtually all of the virus was retained within the capsules.
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  • 208
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 34 (1989) 
    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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  • 209
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    Biotechnology and Bioengineering 34 (1989), S. 1127-1132 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Process scaleup for stirred-tank animal cell cultures such as suspension and microcarrier cultures often begins at the bench scale in small spinner vessels. In order to initiate process development under the proper conditions, it is essential to know the physical conditions under which the cells are grown. In this article, power inputs and surface oxygen transfer rates to culture medium in 500-mL Corning spinner vessels were determined as a function of the impeller geometry, impeller height, and agitation speed. The results obtained indicate that power dissipation dependency differs from literature correlations and may compromise scale up at constant power input from these vessels. These results are of general utility to researchers using small-scale vessels.
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  • 210
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 34 (1989), S. 1133-1139 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Protein localization in Saccharomyces cerevisiae was studied with two plasmid systems used as a model: one containing the SUC2 structural gene fused with the MFα1 (α-factor) promoter and signal-sequence, the other containing the entire SUC2 gene. Special emphasis was placed on the effect of promoter/signal-sequence (SUC2 vs. MFα1) on the efficiency of invertase transport. The MFα 1 and SUC2 signal sequences were capable of transporting, respectively, 83% and 77% of cloned invertase out of the cytoplasm. However, the SUC2 promoter was easier to control since a six-fold enhancement of the transported invertase activity associated with derepression was achieved in response to a glucose concentration change from 10 to 2 g/L Cloning on a multicopy plasmid resulted in a four-fold increase in total specific invertase activity over the wild type yeast strain (which harbors a single copy of the SUC2 gene on the chromosome), whereas the chromosomal site was more efficient for invertase localization yielding over 90% of the invertase transported out of the cytoplasm. Transient experiments done with the SUC2 signal-sequence-containing plasmid showed that the specific invertase activity in the periplasmic space reached a maximum three hours after derepression, then decreased very slowly with an accompanying gradual increase in invertase activity in the growth medium.
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  • 211
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 33 (1989) 
    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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  • 212
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    Biotechnology and Bioengineering 33 (1989), S. 26-31 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The conditions for the introduction of active carbonate groups into supports containing hydroxyl groups by reaction with 5-norbornene-2.3-dicarboximido carbonochloridate are described. Up to 1.5 mmol carbonate groups/g dry Sepharose 4B could be bound. In the case of glycine the reaction of the activated supports with the amino groups takes place with a 10-fold higher rate than the hydrolysis of the carbonate groups, and high coupling yields can be reached. It is shown that the activated supports are well suitable for the preparation of carriers for affinity chromatography or the immobilization of enzymes.
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  • 213
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    Biotechnology and Bioengineering 33 (1989), S. 139-148 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The fluidized-bed, granular activated carbon (GAC) anaerobic reactor has been shown to be an effective process for the continuous long-term treatment of wastewaters that contain biodegradable or nonbiodegradable toxic organic compounds. With loadings of 10 g COD/kg GAC day, COD removal of 94% was achieved. The anaerobic biofilm that develops on the GAC reduces the load on the carbon by converting the biodegradable organics to methane and carbon dioxide. Approximately 50% of the COD applied to the reactor was converted to methane, thereby reducing carbon requirements. Successful operation of the system requires that a carbon replacement schedule be maintained that will keep the bulk concentrations of toxic adsorbable compounds below their toxic threshold. As long as toxic substances can be adsorbed by the carbon, they will not inhibit the anaerobic biofilm. If nonadsorbable toxic compounds are present, processing must be included to reduce these materials to concentrations below their threshold level.
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  • 214
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    Biotechnology and Bioengineering 33 (1989), S. 1-10 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The optimal glucose feeding policy for the fed-batch culture of Saccharomyces carlsbergensis is presented. The biphasic nature of growth results in a singular feed rate policy that is unique to this organism. When the operating cost is high, the reduction in operating time forces the cells to utilize both glucose and ethanol toward the end of fermentation time and results in a decreasing rate of glucose addition, unlike the normally observed in creasing feed rate. The optimal feeding policy depends heavily on the initial conditions and is highly sensitive to changes in kinetic parameters. A semiempirical scheme for feedback optimization is suggested for the fed-batch yeast culture.
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  • 215
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    Biotechnology and Bioengineering 33 (1989), S. 11-15 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Determination of the optimal feed rate for fed-batch fermentation is normally a problem in singular control with a state inequality constraint and as such is, in general, difficult to solve, especially for those described by a large number of dynamic mass balance equations. In this article we use a new set of state variables and the culture volume as the control variable. In this way the problem is converted to one of nonsingular control with the magnitude and rate constraints on the manipulated variable and can be numerically solved by a gradient-based technique, thus avoiding the difficulty associated with singular control problems. Examples are given to illustrate the method.
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  • 216
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    Biotechnology and Bioengineering 33 (1989), S. 72-78 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A bacterial strain, Bacillus licheniformis, has been isolated and identified which produces high-temperature alkaline α-amylase. Cultural conditions, such as types of carbon and nitrogen sources, temperature, pH, and time of reaction, have been optimized for production of α-amylase in shake flask and fermenter. The enzyme produced was quite active even at 100°C; however, it showed optimum activity at 90°C. It exhibited optimum activity in the broad pH range 5.5-10. The effects of Na+ and Ca2+ ions on enzyme activity was also studied.
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  • 217
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    Biotechnology and Bioengineering 33 (1989), S. 95-103 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The breakage of nylon membrane microcapsules is proposed as a new method to study and quantify shear effects in biological systems. A critique of this method shows that a narrower particle size distribution may be an important improvement in the breakage study as well as breakage control in many bioreactor and biotechnological applications. In a turbine reactor, it was shown that the primary process which determines the microcapsule breakage is the shear effect. The breakage kinetics are first order with regard to the microcapsule concentration. The breakage kinetic constant was ob served to be dependent on the temperature and the particle size, and proportional to the average shear rate and the third power of the turbine angular velocity. Decrease of the breakage kinetic constant with temperature can be explained by a decrease of fluid viscosity and a change in nylon membrane properties. An increase in the breakage kinetic constant with the microcapsule diameter can be due to a lowering of internal pressure and a reduction of the membrane resistance with size. Proportionality between the breakage kinetic constant and the shear rate shows that shear is the main process which leads to microcapsule breakage. The additional intervention in the shear rate expression of the turbine angular speed in the form of the turbine and particle velocities, results in the dependence of the breakage kinetic constant on the third power of the angular velocity.
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  • 218
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    Biotechnology and Bioengineering 33 (1989), S. 129-133 
    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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  • 219
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    Biotechnology and Bioengineering 33 (1989), S. 149-156 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We investigated the ability of several hydrolases to catalyze reactions with an abiotic water-insoluble substrate, carbonic acid diphenyl ester, also known as diphenyl carbonate (DPC). In single-phase water/organic systems, turnover numbers (TN) of greater than 2 × 104 min-1have been achieved for the hydrolysis of DPC. The Km values for the hydrolytic reaction were measured to be 200μM and 330μM for Candida cylindracea lipase and Porcine liver esterase, respectively. In addition to hydrolysis, we observed transesterification of carbonates with a wide variety of alcohol and phenol species. Transesterifications of DPC with bifunctional alcohols resulted in the synthesis of polycarbonates. We investigated the stability and transesterification activity of these enzymes in several water-restricted environments to limit competing hydrolysis reactions. We find that, with the removal of water, hydrolysis is reduced more than four orders of magnitude while transesterification is diminished only 10-fold (turnover numbers of 600 min-1 in water-miscible systems to 60 min-1 in water-restricted environments with pure Candida lipase). Stability of the Candida lipase in these water-restricted environments (half-life of longer than 3 days) is much greater than in water/organic single phase systems (5 h in 20% methanol). In addition, the Candida lipase displayed enantiomeric selectivity in transesterifications of DPC with racemic 2-butanol (greater than 80% ee).
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  • 220
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    Biotechnology and Bioengineering 33 (1989), S. 183-190 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Shearing experiments were conducted in a stirred tank reactor with 0.1% lipase solutions of Candida cylindracea. Inactivation of the lipase solutions were observed at various shear rates from 50 to 150 s-1 after continuous shearing for ca. 30-240 min under optimal pH and temperature conditions. However, there was no shear stress denaturation of the lipase when it was subjected to shear stresses of 0.72-109.2 kg/m/s2 and shear rate of 100 s-1. In the presence of polypropylene glycol, the rate of denaturation of the lipase decreased by 93%. When the lipase solution was filled to the brim, the rate of denaturation of the lipase decreased by 97% compared to that when reactor was half-filled. The rate of denaturation of the lipase decreased by 61% when probes in the fermentor were removed. There was no significant difference in the rate of denaturation of the lipase under ambient conditions compared with that in the absence of oxygen, or in the absence of free metal ions. Recovery of lipase activity from the first hour of shearing was observed at a shear rate of 150 s-1. The native lipase and the lipase which had recovered its activity showed similar pH profiles, temperature profiles, and activation energies. Temperature was found to have no effect in the rate of shear-induced denaturation of the lipase in the range 20 to 30°C during shearing at 100 s -1and optimal pH. Above 30°C, the rate of denaturation of the lipase increased drastically as a function of temperature. The significance of the findings in the de sign of reactor systems for hydrolysis or esterification of oils by lipase will be discussed.
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  • 221
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    Biotechnology and Bioengineering 33 (1989), S. 216-220 
    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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  • 222
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    Biotechnology and Bioengineering 33 (1989), S. 229-232 
    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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  • 223
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    Biotechnology and Bioengineering 33 (1989), S. 251-255 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The estimation of the size of each reactor of a series of CSTR's performing a Michaelis-Menten reaction in the liquid phase can be obtained to advantage via an optimization technique leading to the minimum overall capital cost. The cost scaleup is assumed to be described by a power rule on the equipment capacity. Various contributions are lumped into the exponent, thus leading to values above unity. The analytical development leading to the optimal intermediate concentrations of substrate according to the foregoing criterion is presented. A short-cut method based on an empirical expression that approximates the numerical solution is reported. This correlation is found to be exact at the asymptotic behaviors, and to give accurate results within an acceptable error level for the range with physical interest. Therefore, it is particularly useful during the predesign steps of equipment for the biochemical industry.
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  • 224
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    Biotechnology and Bioengineering 33 (1989), S. 321-326 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A successful procedure for attaching cellulase to 45 μm iron oxide particles with a high-molecular-weight (1000) ligand has been developed. Mass and activity balances were calculated for immobilized enzymes with different loadings. The highest specific activity of the immobilized enzyme was 5.9 mmol glucose/g bound protein/h. The efficacy of retaining enzymatic activity was 128%. The optimum pH was 5.5 compared to 4.0 of the free enzyme. The half-life of the IMC was extended to 272 h compared to 0.77 h of the free enzyme.
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  • 225
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    Biotechnology and Bioengineering 33 (1989), S. 344-353 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Enzymatic peptide synthesis was investigated using carboxypeptidase Y immobilized with glutaraldehyde on 10 μm microparticulate amino-silica. Carboxypeptidase Y was immobilized with 98.5% recovery of active enzyme to yield the immobilized enzyme having 0.55 units esterase activity/mg amino-silica support. The stability of the immobilized enzyme was examined as a function of pH, temperature, and reactant concentrations. Immobilized Carboxypeptidase Y was used in stirred batch and recirculating packed-bed reactors for peptide synthesis. Packed-bed reactors (40 × 4.6 mm, 60 × 4.6 mm) were used to catalyze the synthesis of 170 mg N-benzoyl-L-arginyl-L-methioninamide, 380 mg N-benzoyl-L-arginyl-L-methionyl-L-leucinamide, and 200 mg N-benzoyl-L-arginyl-L-methionyl-L-leucyl-L-phenylalaninamide in 8, 3, and 1 hour, respectively, as intermediates in the synthesis of L-methionyl-L-leucyl-L-phenylalanine. No inactivation of the immobilized enzyme was observed during the course of the reactions. The N-benzoyl-L-arginyl group served to increase the water solubility of the peptides and was removed by immobilized trypsin at the end of synthesis to obtain the final product. While the first two syntheses were conducted with aqueous reaction mixtures, the synthesis of N-benzoyl-L-arginyl-L-methionyl-L-leucyl-L-phenylalaninamide was carried out in a reaction mixture containing dimethylformamide to avoid precipitation of the product. HPLC and amino acid analysis confirmed the high purity and amino acid composition of the final product.
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  • 226
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    Biotechnology and Bioengineering 33 (1989), S. 365-368 
    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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  • 227
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    Biotechnology and Bioengineering 33 (1989), S. 1221-1234 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The relationship among extent of hydrolysis, reaction time, and enzyme dosage was investigated. For this, Sigmacell 50 and pretreated poplar wood (20 g/L) was hydrolyzed with varying dosages of cellulases from three different sources (5 to 100 FPU/g) for time periods ranging from 2 to 94 h. It was found that the formation of glucose can be described by summation of two parallel first order reactions. The extent of hydrolysis at fixed time increases with increasing enzyme dosage in a hyperbolic function. From the empirical data it is possible to calculate the fractions of easily and difficult hydrolyzable cellulose and the digestability which could maximally be obtained at infinite enzyme loadings. In the system Sigmacell 50 and Celluclast the easily and difficult hydrolyzable components are 43.0 and 57.0%, respectively, and the maximum digestability at 94 h is 82.6%. Poplar wood, steam treated at 200°, 220°, and 240°C, showed with Celluclast at 24 h a maximum digestability (weight percentage of wood degraded to glucose) of 43.9, 64.9, and 68.0%. The relationships derived from experimental data allow one to compare objectively the effectiveness of different cellulase enzymes and different pretreatments.
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  • 228
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    Biotechnology and Bioengineering 33 (1989), S. 570-577 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The construction and use of an automatic on-line titration unit for routine or event- initiated monitoring of alkalinity, buffer capacity, and volatile fatty acid (VFA) levels is presented. Under computer control a sample of digester liquor is pumped into the titration vessel and weighed. A sequence of titration, sparging, and back-titration operations are then initiated during which the pH and weight are recorded continuously and a titration curve constructed. From the curve, estimates of the alkalinity, buffer capacity to any desired pH endpoint, and total VFA levels are computed. The data is stored to disk and output as hard copy together with the titration curve itself. Monitoring and control of the titration apparatus is effected by a microcomputer via two analog input lines and eight digital output lines, respectively. The system is suitable for downloading to a small, inexpensive dedicated microprocessor-based system. The apparatus is constructed from standard and widely available equipment and the titration sequence, being under software control, is fully adaptable to particular requirements. The use of this facility in the on-line monitoring, control and optimization of the anaerobic digestion process is discussed.
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  • 229
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    Biotechnology and Bioengineering 33 (1989), S. 584-591 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article presents a calculation procedure useful for the optimization and scale up of batch sterilization cycles in large-scale fermentors. This technique determines the sterilization temperature and hold-time necessary to minimize nutrient damage in a specific fermentor. The method can also be used for “scaledown” experiments to eliminate sterilization conditions as a scale up parameter. A method for the systematic evaluation of different sterilization conditions on product yield is also presented. This procedure is useful in determining if scale up of sterilization conditions is important for a given process. The validity of the techniques presented are supported by data showing significant yield improvements in a 1.2 × 105 L antibiotic fermentation.
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  • 230
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    Biotechnology and Bioengineering 33 (1989), S. 1347-1349 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 1 Ill.
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  • 231
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    Biotechnology and Bioengineering 33 (1989) 
    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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  • 232
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    Biotechnology and Bioengineering 33 (1989), S. 1358-1362 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 5 Ill.
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  • 233
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    Biotechnology and Bioengineering 33 (1989), S. 1350-1352 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Acetic acid at concentrations as may occur during vinification and other alcoholic yeast fermentations induced death of glucose-grown cell populations of Saccharomyces cerevisiae IGC 4072 at temperatures at which thermal death was not detectable. The Arrhenius plots of specific death rates with various concentrations of acetic acid (0-2%, w/v) pH 3.3 were linear and could be decomposed into two distinct families of parallel straight lines, indicating that acetic acid induced two types of death: (1) High enthalpy death (HED) predominated at lower acetic acid concentrations (〉 0.5%, w/v) and higher temperatures; its enthalpy of activation (ΔH≠) approached that of thermal death (12.4 × 104 cal/mol); (2) Low enthalpy death (LED) predominated at higher acetic acid concentrations and lower temperatures with ΔH≠ of 3.9 × 104 cal/mol. While the ΔH≠ values for HED induced by acetic acid were similar with those reported earlier for HED induced by other fermentation endproducts, the values for the entropy coefficients were different: 127-168 entropy units mol-1L for acetic acid as compared with 3.6-5.1 entropy units mol-1L for ethanol, which agreed with experimental results indicating that acetic acid is over 30-times more toxic than ethanol with respect to yeast cell viability at high process temperatures.
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  • 234
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    Biotechnology and Bioengineering 33 (1989), S. 1393-1399 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article treats the basic problem of selection of experimental conditions for microbiological experiments for evaluation of newly isolated bacterial strains, mutants, or plasmid/strain combinations. For this purpose shake flask experiments in a 210-4confounded factorial design at resolution IV with four blocks of 16 flasks were used. The design was used for testing of two new strain/plasmid combinations (E. coli MT 102/403-SD2 and W 3110/403-SD2) i.e., both strains with the same plasmid 403-SD2. Both strains were integrated in the design, so both strains were tested with nine factors (temperature, aeration, glucose, initial pH, pH regulation, reduced aeration, chloramphenicol, acetate, and glycerol). With both strains the interaction between initial pH and reduced aeration had a significant influence on the yield of the recombinant-DNA product nuclease. There was more than a factor of 10 between lowest and highest yield of product. In this interactive system the strains reacted differently. MT 102/403-SD2 had highest yields at high initial pH (8.4) and no reduction in aeration, whereas W 3110/403-SD2 had highest yields of nuclease at low initial pH (7.4) and reduced aeration (rubber stopper inserted after cultivation for 12 h). These data (and previous work) clearly demonstrate that it is impossible to suggest a simple set of experimental conditions for testing of new plasmid/strain combinations. It is clear that the exclusive application of a standardized growth technique e.g., LB-medium at 37°C at an unspecified and uncontrolled aeration level, may lead to wrong conclusions on properties and potentials of now plasmid/strain combinations and may lead to rejection of useful strains or plasmids.
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  • 235
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    Biotechnology and Bioengineering 33 (1989), S. 1419-1424 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An adaptive control algorithm for the on-line determination of optimal temperature or pH for biomass production in a continuous fermentor is presented. The algorithm requires no prior information and uses a dynamic Hammerstein model to identify parameters and to estimate an optimal steady-state control value. A check of the estimated performance measure second derivative is included to ensure that the target extremum is an optimum. The process is driven towards this optimum with a variable step size that depends on the quality of the on-line identified model. Numerical simulations are performed on a dynamic chemostat model that incorporates a metabolic time delay. The algorithm successfully finds the optimum temperature or pH values and maintains the reactor at the optimum steady state.
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  • 236
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Gluconobacter suboxydans IFO 3290 was immobilized by adsorption on ceramic honeycomb monolith and continuous production of free gluconic acid from glucose was performed in an aerated reactor. The effects of reactor residence time, aeration rate, and glucose concentration were investigated on the gluconic acid yield. Observation of SEM photographs revealed that the cells were adsorbed with a high density not only on the outer surface of the support but also on the inner surface of large pores. From measurement of the number of the adsorbed cells, it was elucidated that the biofilm comprised a monolayer or bilayer of the cells. Maximum specific rate of growth was estimated for the free and adsorbed cells, and the adsorbed cells were found to grow at a fast rate compared with the free cells. In the continuous fermentation performed for one month at the glucose concentration of 100 kg/m3, reactor residence time of 3.5 h and aeration rate of 900 cm3/min, the activity of the adsorbed cells was appreciably stable. The high productivity of 26.3 kg/(m3-reactor · h) was attained with the gluconic acid yield of 84.6% and glucose conversion of 94%.
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  • 237
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    Biotechnology and Bioengineering 34 (1989) 
    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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  • 238
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    Biotechnology and Bioengineering 33 (1989), S. 631-637 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Adsorption reversibility and competition between fractionated components of the Trichoderma reesei cellulase system were studied. Specific endoglucanase (EGI), nonspecific endoglucanases (EGII, EGIII), and cellobio-hydrolase (CBHI) were previously grouped according to their hydrolytic function. At 5°C, direct evidence of exchange between adsorbed and free enzyme was obtained for each component using [3H] and [14C] radiolabeled tracers. No release of bound enzymes was detected upon dilution of the free enzyme solution. In simultaneous adsorption of enzyme pairs, CBHI was shown to predominate adsorption. Endoglucanase EGI was preferentially adsorbed over EGII and EGIII. Sequential adsorption studies have shown that interaction between enzyme components largely determines the degree of their adsorption. Evidence suggests that both common and distinct adsorption sites exist and that their occupation depends on which components are involved. Predominance in adsorption by any one of the enzyme components is decreased at 50°C. Light microscopy and monitoring of sugar production during cellulose hydrolysis provided evidence that reduction in the ionic strength decreases the adsorption predominance of CBHI and enhances the synergism between the cellulase components.
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  • 239
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    Biotechnology and Bioengineering 34 (1989), S. 160-170 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The authors' mathematical model of transient immobilized cell growth and product formation is applied here to examine the performance of an immobilized cell system subject to periodic cycling of the rate-limiting substrate supply. The model system consists of a single hydrogel-like (porous) particle entrapping viable microorganisms. Proper nutrient cycling is shown to yield a relaxed periodic system and to virtually eliminate the leakage of biomass from the support that is commonly observed experimentally in steady (continuous nutrient supply) operation of these systems. The use of cyclic operation is evaluated by calculating the average product yield (the ratio of product formed to substrate consumed) and the average product flux from the particle (a measure of the total productivity of the system), for various cycling rates. Cycling increased the average product yield by at least a factor of three in nongrowth-related fermentations, relative to steady operation, without any significant sacrifice in average total productivity. Growth-related fermentations lost significant total productivity under most cycling conditions, while the average product yield was approximately unchanged at all cycling rates. Thus, immobilization in conjunction with periodic operation should be considered as an alternative process design for the production of nongrowth-related products such as penicillin and monoclonal antibodies.
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  • 240
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    Biotechnology and Bioengineering 34 (1989), S. 171-179 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Ultrafiltration through Carbosep M4 mineral membrane of protein solutions of decreasing complexity (whey before and after centrifugation or clarification, β-lactoglobulin) was studied. Mathematical models were used to explain variations in flux with time. Taking into account variations in protein retention and hydraulic resistance of the membrane during ultrafiltration, proteins and lipoproteins were found to be involved not only in the polarization layer (reversible fouling leading to a difference in the osmotic pressure), but also in irreversible fouling by adsorption. Morever, the presence of particles (e.g., inorganic precipitates) in whey explains the build-up of a deposit over and within the membrane which contributes to the decline in flux after 1 h ultrafiltration. The relative importance of these phenomena was quantified.
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  • 241
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    Biotechnology and Bioengineering 34 (1989), S. 196-206 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Aiming to satisfy the need for the formalization of semiquantitative reasoning in the analysis of biochemical systems, the O[M] formalism for reasoning with orders of magnitude and approximate relations has been developed. It is based on seven primitive relations among quantities and compound relations which are formed as implicit disjunctions of primitives. O[M] can perform inferences by using formal approximate relations, algebraic equations, inequalities, if-then rules, assumptions, and goals. The applications discussed include Michaelis-Menten kinetics, different modes of inhibition of an enzymatic reaction, analysis of fluxes in biochemical networks, and identification of rate-limiting steps of biochemical pathways. In these applications, O[M] provides a medium for acquisition and formalization of previously informal concepts, analysis of systems at the order-of-magnitude level of detail, and automation of commonsense reasoning.
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  • 242
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    Biotechnology and Bioengineering 34 (1989), S. 242-251 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The behavior of a packed-bed reactor for enzymatic hydrolysis of pretreated wheat straw has been described by means of a mathematical model. The flow pattern has been evaluated by residence time distribution experiments. Small deviations from ideal plug flow behavior were found using the dispersion model. The kinetic model proposed for enzymatic hydrolysis of cellulosic fraction of pretreated wheat straw has been derived from batch experimental data. Variations of enzyme concentration throughout the straw bed have been approximately described using a ramp variation of adsorbed enzyme. The final explains qualitatively the experimental results.
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  • 243
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    Biotechnology and Bioengineering 34 (1989), S. 214-222 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Plant cells of Mucuna pruriens L. entrapped In calcium alginate, calcium pectinate, agarose, or gelatine were able to convert L-tyrosine to L-DOPA, which was released Into the medium. Michaelis-Menten kinetics could be applied on the entrapped cells, based on the measurement of initial rates of L-DOPA production. The calculated apparent affinity constants were comparable with the affinity constants obtained with enzyme preparations. Comparison of the apparent maximum rate of bioconversion of the entrapped cells and the maximum rate of bioconversion of a derived cell homogenate indicated that the systems were not operating optimally. Measurement of the effective diffusion coefficients of L-tyrosine pointed out that this substrate could diffuse freely into the matrices. From the initial rates of bioconversion and the effective diffusion coefficients, the observable modulus was calculated for each system. The obtained values confirmed that the diffusional supply rate of L-tyrosine was not the limiting factor. For oxygen, which was utilized for byconversion as well as for cell respiration, the calculated observable moduli was directed toward strong oxygen transfer limitations. The values found for the oxygen consumption indicated that the entrapped cells remained partly or totally viable in the four matrices tested. Based on the highest viability and the highest rates of bioconversion, it was concluded that alginate-entrapped cells of M. pruriens formed the most suitable biocatalytic system for the production of L-DOPA from L-tyrosinre.
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  • 244
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    Biotechnology and Bioengineering 33 (1989), S. 1385-1392 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Penicillin was recovered from mycel-containing fermentation broth by direct reactive extraction into a counter-current extraction decanter, Type CA 226-290 of the Westfalia Separator Co., at room temperature via steady state operation. Penicillin concentrations in the feed varied from 3 to 41 g L-1, Amberlite LA-2 carrier concentrations from 7 to 20 g L-1and/or DITDA carrier concentrations from 7.2 to 84 g L-1, the LA-2-to-penicillin mole concentration ratio from 4 to 6.4, and/or the DITDA-to-penicillin mole concentration ratio was maintained at 2. The throughputs of the fermentation broth (520 to 880 L h-1) of the solvent phase (200 to 860 L h-1) and the over all throughput (800 to 1750 L h-1) were high. Extraction degrees of 72 to 96% were achieved between pH 4.6 and 5.1. Without carriers in the same pH range, extraction degrees of only 17 to 19% were attained. By reducing the pH to 2.3 and in the absence of carriers, the degree of extraction was increased to 61%. However, during the extraction, 6.5% of the penicillin decomposed. At these high throughputs, the steady state was attained within 1 to 4 min. Through the mechanical stress, the length of the hyphae was reduced and the protein content of the broth was increased by 50 to 100%. However, this protein content had no appreciable influence on the phase separation.
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  • 245
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    Biotechnology and Bioengineering 33 (1989), S. 1379-1384 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Acoustic resonance densitometry (ARD) provides a highly reproducible and stable method for on-line measurement of culture biomass density. The technique provides a direct determination of changes in relative density of culture medium and cell mass. At cell concentrations higher than 106 cells mL-1this method can replace cell counts and provide a continuous measure of total cell mass. In cultures of hybridomas or U937 human lymphoma cells, the ARD value correlates well with cell number except when the average cell size changes during culture. It is argued that cell mass determined by ARD rather than cell number should be used as the basis for measurements of specific biological activity.
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  • 246
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    Biotechnology and Bioengineering 33 (1989), S. 1445-1460 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A previously isolated cellodextrin glucohydrolase (β-glucosidase) from Trichoderma reesei QM 9414 is characterized using β-1,4-glucose oligomers with defined degrees of polymerization as soluble substrates. The enzyme splits off glucose units from the nonreducing chain ends of cellooligomers. Besides this hydrolytic activity there is also evidence for transfer activity depending on the concentration and degree of polymerization of substrates. Concentration-time-course data have been gathered for the degradation of cellobiose, cellotriose, cellotetraose, cellopentaose, and cellohexaose covering a wide range of enzyme and substrate concentrations. A Michaelis-Menten type kinetic model has been developed, which is able to satisfactorily describe the complex system of parallel and series reactions during the conversion of oligomers to glucose. The only kind of inhibition considered is competitive inhibition by the final product glucose. The model takes into account the formation of multiple enzyme-substrate complexes and is limited to those conditions, in which no transglucosylation products are observed. Cellodextrins with higher degrees of polymerization are found to be better substrates for this enzyme than is the dimer cellobiose.
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  • 247
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    Biotechnology and Bioengineering 33 (1989), S. 1477-1481 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The water-in-oil microemulsion system bis(2 ethyl-hexyl-sodium-succinate (AOT)/isooctane/water is able to solubilize soybean nodules mitrochrondria. Transparent and thermodynamically stable hydrocarbon solutions are obtained, which can be assayed for mitochondrial activity just as aqueous solutions. Malate dehydrogenase (MDH) activity was measured in vivo and gave in reverse micelles very similar results as in water. However the kinetic behavior of this reaction in AOT/isooctane reverse micelles shows some differences with respect to water. Mitochondria in reverse AOT micelles are able to retain about 70% of their initial MDH activity after three days. Mitochondria can be back-transferred from reverse micelles to water and show respiratory activity almost identical to the native organelles. Electron microscopy studies show that the dimensions of mitochondria back-transferred into water from AOT micelles are comparable to the dimensions of the native organelles.
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  • 248
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    Biotechnology and Bioengineering 34 (1989), S. 689-693 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Near-homogeneous forms of glucoamylases I and II, previously purified from an industrial Aspergillus niger preparation, were used to hydrolyze a number of di- and trisaccharides linked by α-D-glucosidic bonds. Maximum rates and Michaelis constants were obtained at various temperatures and pH values with glucoamylase I for the disaccharides β,α-trehalose, kojibiose, nigerose, maltose, and isomaltose and the trisaccharides panose and iso-maltotriose, and with glucoamylase II for maltose, maltotriose, and isomaltotriose. Maximum rates were highest and energies of activation were lowest for maltose, maltotriose, and panose, the only three substrates containing α-D-(1, 4)-glucosidic bonds. Michaelis constants were lowest and standard heats of binding were most negative for maltose and maltotriose. The variation of maximum rates and Michaelis constants with varying pH values suggested that two carboxyl groups were involved in substrate binding.
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  • 249
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    Biotechnology and Bioengineering 34 (1989), S. 705-716 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A wild-type strain of Klebsiella oxytoca growing aerobically in batch culture has exhibited intermittent or oscillatory growth while growing on lactose at concentrations on the order of 1 g/L or less. In two-substrate experiments, preferred growth on glucose followed by growth on lactose also produced oscillatory growth behavior during the lactose growth phase at lactose concentrations of 1 g/L or less. Only oscillations in cell density have currently been observed. Alkalinization of the medium during growth on lactose indicated the presence of lactose active transport. The observed intermittent growth was reduced or removed during growth on lactose after preferred growth on galactose or in a medium containing 50 mM NaCl. Results suggested that the presence of an intracellular energy source or a sufficient ΔpH buffer may alleviate growth inhibition when transport and growth processes compete for essential energy resources during growth on lactose.
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  • 250
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    Biotechnology and Bioengineering 34 (1989), S. 10-17 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The potential of uranium recovery from the dilute uranium ore bioleach solutions of the Elliot Lake district of Canada was examined using immobilized microbial biomass. Batch and continuous laboratory scale pilot plant experiments were carried out. The results have shown that the immobilized microbial biomass can successfully recover all of the uranium from dilute (less than 300 mg U/L) solutions. The uranium can subsequently be eluted producing a high uranium concentration eluate perhaps exceeding 5000 mg U/L. The biomass maintained its biosorption capacity of about 50 mg U/g over 12 examined successive adsorption-elution cycles with no apparent indication of failure.
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  • 251
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    Biotechnology and Bioengineering 34 (1989), S. 804-818 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A pH-dependent “series-type” enzyme deactivation model using rapid protonation and deprotonation equilibria and the relatively slower inactivation rates is presented. From the enzyme activity-time trajectories at different pH the models presented permit the evaluation of some of the protonation and inactivation rate constants as well as the specific activities of the different enzyme forms. pH dependence of enzyme deactivations may also exhibit deactivation disguised kinetics. Three different examples of pH-dependent enzyme deactivations available in the literature are appropriately modeled to indicate the general applicability of the model. The model presented is consistent with the data and provides mechanistic insights into the pH-dependent deactivation of different enzymes.
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  • 252
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    Biotechnology and Bioengineering 34 (1989), S. 59-64 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Cellulomonas sp. ATCC 21399 produced extracellular enzyme activities against Avicel, H3PO4-swollen Avicel, carboxymethylcellulose, (1-3, 1-4)-β-D-heteroglucan, xylan, galactomannan, and amylose drying growth on microcrystalline cellulose. No extracellular cellobiase activity was produced. Crossed immunoelectrophoresis of the crude extracellular enzyme system revealed 15 immunologically distinct immunoprecipitates. The immunoprecipitates of endoglucanase A, endoglucanase B and the xylanase appeared heterogeneous with several optima, whereas the immunoprecipitates of endoglucanase C and the amylase appeared homogeneous. The heterogeneity of endoglucanase A, endoglucanase B and xylanase was also visualized using electrofocusing-immunoelectrophoresis. Electro-focusing could resolve the activity against carboxymethylcellulose into six peaks, whereas only one peak of activity against Avicel was observed. The later peak coincided with the major peak of activity against carboxymethylcellulose with isoelectric point between pH 4.0-5.0.
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    Biotechnology and Bioengineering 34 (1989), S. 577-591 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A multivariable on-line adaptive optimization algorithm using a bilevel forgetting factor method was developed and applied to a continuous baker's yeast culture in simulation and experimental studies to maximize the cellular productivity by manipulating the dilution rate and the temperature. The algorithm showed a good optimization speed and a good adaptability and reoptimization capability. The algorithm was able to stably maintain the process around the optimum point for an extended period of time. Two cases were investigated: an unconstrained and a constrained optimization. In the constrained optimization the ethanol concentration was used as an index for the baking quality of yeast cells. An equality constraint with a quadratic penalty was imposed on the ethanol concentration to keep its level close to a hypothetical “optimum” value. The developed algorithm was experimentally applied to a baker's yeast culture to demonstrate its validity. Only unconstrained optimization was carried out experimentally. A set of tuning parameter values was suggested after evaluating the results from several experimental runs. With those tuning parameter values the optimization took 50-90 h. At the attained steady state the dilution rate was 0.310 h-1 the temperature 32.8°C, and the cellular productivity 1.50 g/L/h.
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  • 254
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    Biotechnology and Bioengineering 34 (1989), S. 660-670 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Model experiments were performed with starved yeast (Saccharomyces cerevisiae) cultures in a batch reactor in order to develop a better understanding of NAD(P)H and flavin culture fluorescence. Fluorescence was monitored during aerobic-anaerobic-aerobic transitions and ethanol and glucose substrate addition experiments. Interpretations of the fluorescence responses obtained are provided, with consideration given to redox compartmentation and the formation of ethanol shortly after a glucose addition. An analytical spectrofluorophotometer was interfaced to a personal computer and adapted to measure fluorescence in a bioreactor. This was achieved by the use of quartz fiber-optic waveguides to convert the right-angle cuvette geometry of the analytical spectrofluorophotometer to an open-ended fluorescence probe geometry, resulting in a flexible culture fluorescence apparatus. Features of the apparatus include variable excitation and emission wavelengths, allowing for detection of NAD(P)H or flavin fluorescence, as well as small slit widths, a variable sampling rate, excitation and emission scanning capabilities, and good sensitivity.
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  • 255
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    Biotechnology and Bioengineering 34 (1989), S. 681-688 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Glucoamylase, industrially derived from Aspergillus niger, was chromatographically separated into forms I and II and purified to near homogeneity. Preparations were proved to be free of D-glucosyltransferase by electrophoretic and differential inhibition tests. Maximum rates and Michaelis constants were obtained for both glucoamylases I and II with maltooligosaccharides from maltose to maltoheptaose and with isomaltooligosac-charides from isomaltose to isomaltohexaose. Subsite maps were calculated from these kinetic data and were not significantly different for the two forms. Subsites in both forms had lower affinities for D-glucosyl residues contained in isomaltooligosaccharides than for D-glucosyl residues in maltooligosaccharides.
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  • 256
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    Biotechnology and Bioengineering 34 (1989) 
    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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  • 257
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    Biotechnology and Bioengineering 34 (1989), S. 750-754 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An oscillatory behavior in population density was observed when a transformant of Bacillus stearothermophilus carrying a rocombinant plasmid pZAM26 was cultivated continuously in a well-stirred reactor vessel at a fixed dilution rato. Among the transformant cells that were subjected to the continuous culture, the fraction of cells harboring p2AM26 was found to be as high as 0.98-1.00 despite the emergence of the oscillation. Cells whose plasmids underwent rearrangement of DMA in terms of structural change could not be found throughout. With reference to this observation, the dynamics of the genetic-engineered bacterium was analyzed within the category of both the linearized stability principle and the bifurcation theory. It was concluded that Hopf bifurcation was most probable to account for the experimental oscillation.
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  • 258
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    Biotechnology and Bioengineering 34 (1989), S. 755-762 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Various water-immiscible solvents were tested for biocompatibility and hydrocarbon recovery under different contact conditions with the hydrocarbon-rich microalga Botryococcus braunii. Eighteen solvents were first selected from a database of 1500 compounds (compiled for solvent selection for ethanol recovery from Saccharomyces cerevisiae fermentation). Nine of these candidate solvents were shown to be biocompatible with B. braunii following short contact times. This biocompatibility tends to be associated with high molecular weights and high boiling points but strongly depends on solvent chemical structure. A low polarity is essential to biocompatibility and calculated octanol-water partition coefficients, or capacity factors determined by reversed-phase high-performance liquid chromatography (HPLC), are suitable predictors of biocompatibility with B. braunii. High recoveries of hydrocarbons directly from the algal culture require relatively polar solvents and are, therefore, inimical with maintenance of cell viability. The inaccessibility of weakly polar solvents to the cell surface appears to protect the algae but also prevents substantial recovery of the hydrocarbons stored in B. braunii outer walls. In order to achieve a high recovery, contact with the solvent must be carried out on algae concentrated by filtration. Then, a large fraction of B. braunii hydrocarbons can be recovered, after a short contact time, without impairing cell viability. Under these conditions, the pertinent solvent property is affinity for the nonpolar hydrocarbons, and the highest recovery yield, ∼70% after contact for 30 min, is achieved with hexane.
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  • 259
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    Biotechnology and Bioengineering 34 (1989), S. 825-837 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The bioconversion activity of calcium alginate-immobilized Clostridium acetobutylicum ATCC 824 was investigated in a continuous reactor system utilizing a defined feed medium which did not support cell growth. The changes in biocatalytic activity with time were studied at different pH values as well as when different metabolites (butyric, acetic, and acetoacetic acids) were present in the feed stream. Although the nongrowing cells were metabolically active, the product distribution was shifted from solvent production to acidogenesis. Overall activity losses occurred due to cell lysis, sporulation, and the effects of nitrogen level on macromolecular turnover. These effects were minimized under some operating conditions (e.g., pH 6), resulting in significantly longer productivity lifetimes.
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  • 260
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    Biotechnology and Bioengineering 34 (1989), S. 863-867 
    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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  • 261
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    Biotechnology and Bioengineering 34 (1989), S. 872-874 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 5 Ill.
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  • 262
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    Biotechnology and Bioengineering 34 (1989), S. 902-908 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Experiments were performed to evaluate, qualitatively and quantitatively, the adaptation of Escherichia coli to plasmid maintenance and cloned gene expression. Experimental findings indicate that the metabolic response to low plasmid levels is an increase of the biosynthetic capacity of both transcription and translation. At high copy number levels the gene-specific transcription rate continues to increase but the stability of plasmid-derived mRNA drops sharply. Protein levels are maintained, but translation efficiency decreases. These results indicate that cellular biosynthetic capacity may not be limiting productivity in recombinant systems. If macromolecular stability is the bottleneck, then current efforts to increase gene expression that focus on enhancing synthesis rates will be ineffective.
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  • 263
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    Biotechnology and Bioengineering 34 (1989), S. 933-941 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Although higher initial rates of phenylacetyl carbinol formation were observed in fermentations containing a high starting benzaldehyde level, a massive reduction in yeast viability was observed resulting in early cessation of production formation. Pulse feeding to maintain lower benzaldehyde concentrations resulted in a lower initial reaction rate, but prolonged yeast viability and the biotransformation. This resulted in higher overall product tilers. As benzaldehyde concentration was increased, yeast growth rate was reduced (0.5 g/L), inhibited (1-2 g/L), or cell viability reduced (3 g/L). Benzaldehyde appeared to alter the cell permeability barrier to substrates and products. Reductions in yeast biomass levels and especially protein and lipid content were observed during the biotransformation. The effects of benzaldehyde and reaction products on yeast pyruvate decarboxylase and alcohol dehydrogenase stability were determined. Homogenized yeast cells produced similar phenylacetyl carbinol levels to whole yeast only if supplemented with thiamine pyrophosphate and magnesium.
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  • 264
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    Biotechnology and Bioengineering 34 (1989), S. 1000-1014 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Phase diagram data at 4°C was determined for the aqueous two-phase systems composed of polyethylene glycol, dextran, and water. The Flory-Huggins theory of polymer thermodynamics was used to correlate partitioning of biomolecules in these aqueous two-phase systems resulting in a simple linear relationship between the natural logarithm of the partition coefficient and the concentration of polymers in the two phases. This relationship was verified by partitioning a series of dipeptides which differ from one another by the addition of a CH2 group on the c-terminal amino acid residue and by utilizing a set of low-molecular-weight proteins. The slope of the line could be expressed in terms of the interactions of the biomolecule with the phase forming polymers and water. The main result for the dipeptides was that knowledge of the partition coefficient in any of the PEG/dextran/water systems, regardless of polymer molecular weight, enabled prediction of the coefficient in all of the systems. The dipeptides were also used for determination of the Gibbs free energy of transfer of a CH2 group between the phases. This quantity was correlated with polymer concentration, thus establishing a hydrophobicity profile for the PEG/ dextran/water systems. The methodology for predicting dipeptide partition coefficients was extended to proteins, where it was found that low-molecular-weight proteins gave a linear relationship with the tie line compositions of a phase diagram.
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  • 265
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    Biotechnology and Bioengineering 34 (1989), S. 1045-1049 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In continuous aerobic treatment of artificial sewage by activated sludge, the rate of removal of chemical oxygen demand (COD) was markedly enhanced by the presence of cross-linked poly-4-vinylpyridine (PVP). The concentration of dissolved oxygen (DO) was also low in the presence of PVP. The extent of improvement in COD removal increased with increases in substrate load and the surface area of the PVP in the working space of the test apparatus. These results suggest an increase of the bacterial population resulting from the presence of the PVP. However, formation of measurable biofilm was not detected on the surface of the PVP during continuous aerobic treatment.
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  • 266
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    Biotechnology and Bioengineering 34 (1989), S. 1058-1062 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of oxygen supply on the growth of suspension cultures of Catharanthus roseus in Erlenmeyer flasks was investigated. Below a critical oxygen supply rate the culture could not survive. By increasing the oxygen supply, a point is reached where the culture survives but no growth is possible. At higher oxygen supply rates there is a regime where both growth rate and the maximum biomass concentration increase with oxygen supply. Eventually there comes a point where no further increase in biomass is achieved, probably due to the depletion of the sugars; however, the growth rate continues to increase with oxygen supply until a maximum growth rate is obtained. The ratio of fresh to dry weight at maximum fresh weight increased with shaker table speed of rotation accompanied by a greater rate of sugar depletion.
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  • 267
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    Biotechnology and Bioengineering 34 (1989), S. 1104-1113 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of forced oscillations in the dilution rate on a population of Escherichia coli K12 harboring the plasmid pBR322 in a chemostat with a nonselective medium were studied. In the constant dilution rate control experiments, the percentage of plasmid-containing cells decreased after a long lag time. Eventually, the culture approached a population consisting of 100% plasmid-free ells. However, under forced perturbations of the dilution rate, the culture maintained a mixed population of plasmid-free and plasmid-carrying cells for a longer period of ime. An unstructured model was developed to describe the above observations. Our results indicate that transient conditions created by dilution-rate perturbations provide a favorable environment for the plasmid-carrying population. In addition, experiments with different cycling frequencies suggest that adaptation by the culture to these transient conditions will reduce or totally eliminate such an advantage.
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  • 268
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    Biotechnology and Bioengineering 33 (1989), S. 906-911 
    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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  • 269
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    Biotechnology and Bioengineering 33 (1989), S. 896-905 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Substrate regulation patterns were changed by covalent binding of bovine liver glutamate dehydrogenase via primary amino groups to CNBr- and CH-activated Sepharose 4B. Lineweaver-Burk plots show that the NAD activation region changed from being abrupt to elongated when the enzyme was immobilized to either support. The elongated region contains two inflection points and resembles substrate activation of several other allosteric oligomers. Glutamate induced varying degrees of abrupt activation in immobilized glutamate dehydrogenase and inhibited the native enzyme. This activation is characterized by an activation threshold, an increase in the apparent dissociation constant, and a correlation between the apparent rate constant and the degree of activation. These three features characterize other glutamate dehydrogenase systems.
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  • 270
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    Biotechnology and Bioengineering 33 (1989), S. 886-895 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An important factor complicating the recovery of recombinant proteins from Escherichia coli is their intracellular location. An alternative to the commonly used method of releasing these proteins by mechanical disruption is to chemically permeabilize the cells. The objective of this research was to characterize the protein release kinetics of a permeabilization process using guanidine-HCl and Triton X100. The protein release rate and yield were determined as a function of the guanidine and Triton concentrations. The initial release rate increased monotonically with increasing concentrations of Triton and guanidine whereas the release yield varied in a complex manner. Electron microscopy indicated that the permeabilization process involves a solubilization of the inner membrane and molecular alteration of the outer wall. Some advantages of this process over mechanical disruption include avoiding extensive fragmentation of the cells and retainment of nucleic acids inside the cell structure.
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  • 271
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    Biotechnology and Bioengineering 33 (1989), S. 873-885 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The hydrolysis of lactose using immobilized β-galactosidase (from Aspergillus niger) on phenol-formaldehyde resin was studied at temperatures between 8 and 60°C and initial lactose concentrations ranging from 2.5 to 20.0%. A model involving enzyme-galactose complex similar to Michaelis-Menten kinetics with competitive product (galactose) inhibition is suitable to describe the lactose hydrolysis reaction. A small degree of lack of fit between the model and the data was found to be due to the formation of oligosaccharides. Thermal deactivation of lactase follows first-order reaction mechanism. The effect of temperature on the reaction and the deactivation rate constants follows the Arrhenius relationship. The Oligosaccharide formation was not significantly affected by the temperature when the initial lactose concentration was 5%. A design equation for the plug-flow immobilized lactase reactor was developed from the reaction and the deactivation kinetics and was used to find the optimal operating temperature. The optimal temperature was found to be dependent on the operating time but not on the lactose concentration or the conversion. The optimal operating temperature is 60°C when operating time is short but is close to 35°C for a long operating time. A preliminary economic analysis indicates that the optimal operating temperature is 43, 38.5, and 33°C when the operating time is 300 days, 1000 days, and infinity, respectively.
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    Biotechnology and Bioengineering 33 (1989), S. 912-914 
    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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  • 273
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    Biotechnology and Bioengineering 33 (1989), S. 915-917 
    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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  • 274
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    Biotechnology and Bioengineering 34 (1989), S. 1295-1304 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The direct microbial conversion (DMC) process for the production of ethanol from lignocellulosic biomass is limited by low volumetric ethanol production rates due to the low cell densities of Clostridium thermosaccharolyticum which is a key organism for ethanol production in this process. Hence, this study focuses on the use of a continuous- culture cell recycle system to improve the volumetric ethanol productivity and yield of the fermentation of xylose by C. thermosaccharolyticum. Early experiments with the continuous-culture cell recycle system showed a two-fold improvement in volumetric ethanol productivity. However, the ethanol yield at the higher dilution rates suffered because of the large amount of lactate produced. The manipulation of two environmental parameters - iron concentration in the nutrient medium and the N2 purge rate of the fermentor headspace - allowed a dramatic reduction in the lactate production and a simultaneous improvement in the ethanol titer and yield. Under the improved conditions of increased iron concentration (12.5 mg/L FeSO4 · 7H2O) and decreased N2 purge rate (0.1 L/min), a continuous culture of C. thermosaccharolyticum operating at a dilution rate of 0.24 h-1 and 50% cell recycle produced 8.6 g/L ethanol and less than 1 g/L each of acetate and lactate. The volumetric ethanol productivity was 2.2 g/L/h, which is 8 times larger than obtained for a continuous culture operated with no cell recycle and the same specific growth rate.
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  • 275
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 5 Ill.
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  • 276
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    Biotechnology and Bioengineering 34 (1989), S. 1321-1325 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The method for purification of biomolecules by a combination of affinity interactions and membrane filtration for separation of unwanted material has been found to be of interest for large-scale work. This study examines the suitability of silica nanoparticles as carriers in the process. Alcohol dehydrogenase and lactate dehydrogenases were chosen as target molecules to be purified. The binding capacity was found to be comparative to what is obtained for high-performance liquid chromatography (HPLC) packing material. Both binding and desorption of the enzymes were found to be effective. The limiting factor of the process was the filtration flow rate.
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  • 277
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    Biotechnology and Bioengineering 33 (1989), S. 948-954 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Direct saccharification of 2.64% cassava starch by Rhizopus oligosporus 145F was attempted under various cultural conditions. Maximum glucose yield of 18.0 g/L culture filtrate was obtained with an initial pH 3.8, 2% (v/v) inoculum of R. oligosporus spores, and an incubation temperature of 45°C in shake flask cultures for 48 h. This concomitantly produced 2.7 g mycelia/100g cassava starch containing 20.2% true protein. The production of glucose and mycelia was accomplished with 92.8% starch saccharification having 67.9% starch to glucose conversion efficiency.
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  • 278
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    Biotechnology and Bioengineering 33 (1989), S. 941-947 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A nondestructive method of measuring extracellular polysaccharides (ECP) in activated sludge floes using Ruthenium Red dye adsorption was developed at the Environmental Engineering Laboratory at the University of Colorado at Boulder. The effects of pH, buffer solution, dye concentration, sludge mass, temperature, and incubation time on dye adsorption was determined. Ruthenium Red dye adsorption to bacterial floes was found to fit a Brunauer-Emmett-Teller (BET) isotherm model. Of the other environmental conditions in the system, pH was found to have the strongest effect on dye adsorption to bacterial flocs. The amount of extra cellular polysaccharides (ECP) measured by Ruthenium Red adsorption was compared with extracellular polysaccharides measured by two chemical extraction methods. Of all methods considered Ruthenium Red dye adsorption measured the highest amount of extracellular polysaccharide with the lowest amount of bacterial cell disruption. Thus, Ruthenium Red dye adsorption was more effective than extraction procedures for measurement of extracellular polysaccharides in activated sludge flocs.
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  • 279
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    Biotechnology and Bioengineering 33 (1989), S. 955-962 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A continuous enzymatic hollow fiber reactor (HFR), obtained by immobilizing cellobiose active cells into the shell side of hollow-fiber modules, was studied. The HFR yield was monitored by glucose analysis resulting from hydrolysis of cellobiose. The residence time of substrate in the bioreactor to obtain convenient hydrolysis yields was calculated from tests carried out by varying the reactor dilution rate in the range 0.001-0.004 L/min. The glucose yield was measured for 300 h (continuous substrate flux). The yield decreased from 40 to 15%. This decrease was due to the loss of specific activity in the operating conditions and to the pressure drop increase from 0.2 to 1.7 atm. The pressure drop increase is in turn dependent on the cell loading (0.2-2.1 g dry cell) and the substrate flux.
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    Biotechnology and Bioengineering 33 (1989), S. 963-975 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A general theory is presented in this article for determining the intrinsic rate constants for the main reaction and deactivation reaction, the effective diffusivity of the substrate, and the active enzyme distribution within porous solid supports from deactivation study of a continuous stirred-basket reactor (CSBR). For the parallel deactivation five reaction kinetics are considered: (a) Michaelis-Menten, (b) substrate inhibition, (c) product inhibition (competitive), (d) product inhibition (anticompetitive), and (e) zero-order kinetics. The experimental results of the system of hydrogen-peroxide-immobilized catalase on controlled-pore glass particles are analyzed to demonstrate the application of the theory developed for parallel deactivation of active immobilized enzyme (IME). For series deactivation only first-order kinetics is treated, and a numerical procedure is proposed to deter mine the rate parameters and the internal active enzyme distribution. The experimental data of the system of glucose-immobilized glucose oxidase on silica-alumina and controlled-pore glass particles are used to verify the theory.
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    Biotechnology and Bioengineering 33 (1989), S. 976-983 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Expression kinetics of the human Epidermal Growth Factor (hEGF) from the α-factor prepro region in a 2-μm based plasmid was studied in Saccharomyces cerevisiae. Production of hEGF was highly medium de pendent as a chemically defined, nonenriched media had a significantly lower yield than did enriched media. Also cells grown on yeast nitrogen base without amino acids with casamino acids degraded the hEGF after cell growth as opposed to a yeast extract, peptone, and dextrose (YEPD) medium, which elicited no measurable extracellular proteolysis of the hEGF. α-factor directed production kinetics of hEGF on the YEPD medium were growth associated, secretion limitations and extracellular degradation were negligible, and the hEGF was nearly 100% selectively secreted. With sufficient agitation, shake flask experiments were representative of aerated controlled batch fermentations. No effect of high cell density was observed on cell growth or hEGF production kinetics. The hollow fiber bioreactor had no direct effect on the substrate or protein yields of S. cerevisiae, however the low oxygen transfer capacity of the membrane was not sufficient to support respiration.
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  • 282
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    Biotechnology and Bioengineering 34 (1989), S. 741-749 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The possibility of enhancing the biomass productivity of a continuous culture of Saccharomyces cerevisiae growing on a glucose-limited medium is addressed. An unstructured Monod-type model is first identified using steady-state data. The culture is subjected to step changes in dilution rate, and it is seen that the Monod model is unable to predict even qualitatively the dynamic response of the culture. Incorporation of a time delay allows significant improvement in the transient fit. It is found that the culture has a time lag of about 3 h in adapting its growth rate. Cycling the dilution rate with a period of 3 h leads to substantial improvement in the average biomass productivity.
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  • 283
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    Biotechnology and Bioengineering 34 (1989), S. 794-803 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The aim of this article is to develop the on-line measurement system of enzyme reaction rates, using an advanced pH controller, which is called a repetitive PF(Programmed Controller/Feedback Compensator) System. In the esterolysis reaction of the N-acetyltyrosine ethyl ester by α-chymotrypsin, the enzyme reaction rate could be calculated from the amount of base required for keeping the pH constant. The proposed controller has a learning mechanism in which the knowledge is obtained from the former results of the repetition, and pH was controlled much more successfully by the proposed repetitive PF System than by the conventional on-off controller, PI controller, and the adaptive controller. The enzyme reaction rate could be evaluated each time as accurately as possible based on the result of the controlled batch reaction. Using Lineweaver-Burk plots of the estimated reaction rates, kinetic parameters, such as the enzyme activity and Michaelis-Menten constant, could be estimated more accurately and from fewer experiments by the repetitive PF system than by initial reaction rates using on-off pH-stat.
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  • 284
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    Biotechnology and Bioengineering 34 (1989), S. 819-824 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The efficiency of two different agitation systems (airlift and paddlewheel) in the biomass photoproduction of a nitrogen-fixing filamentous blue-green alga was evaluated outdoors, and the elemental and molecular composition of the cells grown with each system was analyzed. With the paddlewheel system, the productivity values achieved were over 30% higher than with the airlift system, both in summer and winter. In this last season, a conversion efficiency of total solar energy into stored biomass energy of 3.3% was estimated for the paddlewheel system. Moreover, the algal cells grown with this system exhibited a higher net protein (58.9% of dry weight) and nitrogen (11.3%) content than those grown with the airlift device, with an estimated nitrogen fixation rate of more than 2 g N m-2 day-1. These advantages of the paddlewheel system make this procedure more appropriate for the large-scale photoproduction of nitrogen-fixing blue-green algae outdoors.
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  • 285
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    Biotechnology and Bioengineering 34 (1989), S. 882-884 
    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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  • 286
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    Biotechnology and Bioengineering 34 (1989), S. 885-895 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Energetic analysis was applied to reduced product formation following perturbation of ethanol- and propionate-fed methanogenic continuous stirred tank reactors (CSTRs). Formation and dissipation of longer-chained n-carboxylic acids corresponded with the variation in Gibbs free energy change associated with beta-oxidation reactions. Formation appeared to occur from acetate and propionate by reductive back-reactions, made energetically favorable by elevated hydrogen partial pressure (PH2), and possibly mediated by biosynthetic enzymes. The formed longer-chained acids dissipated when the PH2 fell and equilibrium shifted to favor beta-oxidations. n-Propanol was found to be produced from propionate in a coupled ethanol oxidation/propionate reduction reaction, mediated by ethanol-oxidizing organisms during high rates of ethanol utilization and elevated PH2. When PH2 declined, n-propanol was oxidized back to its precursor propionate. Both reaction energetics and intracellular diffusion of the electron carrier may effect transient mediation of this coupled reaction.
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  • 287
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    Biotechnology and Bioengineering 34 (1989), S. 916-925 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Traditionally, enzyme populations have been treated as if they were either homogenous, or heterogeneous with distinct and separable subpopulations. The microheterogeneity model, however, assumes that there is a continuous distribution of properties in the population. In the area of enzyme deactivation kinetics, this model describes the heterogeneous population as having a continuous distribution of activation energy of deactivation. This distribution is characterized by mean activation energy, and a standard deviation of activation energy. The microheterogeneity model contains two parameters, ∊0 and σ. Parameter ∊0 is the mean value of ∊ for a heterogeneous enzyme population; ∊ is the activation energy divided by absolute temperature and the ideal gas constant. Parameter σ is the standard deviation of the Gaussian distribution of ∊ values in the population. If the population is homogeneous, then ∊ = ∊0 for all enzyme molecules and σ = 0. There are certain ratios which are independent of ∊0 and dependent upon σ. Two important ratios are t1/4/t1/2 and t1/2/t1/2′, where t1/2′ represents t1/2 for a homogeneous enzyme population with the same mean ∊ (∊0), as the heterogeneous population. If there is experimental deactivation data for the heterogeneous population which is well behaved, the first ratio, t1/4/t1/2, can be determined by estimating the time in minutes at which the enzyme has lost 25% of its activity (t1/4), and the time in minutes at which the enzyme has lost 50% of its activity (t1/2), and then taking the ratio t1/4/t1/2. The corresponding value of σ can be estimated from a graph. The ratio t1/2/t1/2′ can be found directly as a function of t1/4/t1/2, and can be estimated from another graph. The value of ∊0 can then be calculated from the formulasgiven in the article.
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  • 288
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    Biotechnology and Bioengineering 34 (1989), S. 926-932 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article proposes a simple steady-state method for measuring the effective diffusion coefficient of oxygen (De) in gel beads entrapping viable cells. We applied this method to the measurement of De in Ca- and Ba-alginate gel beads entrapping Saccharomyces cerevisiae and Pseudomonas ovalis. The diffusivity of oxygen through gel beads containing viable cells was measured within an accuracy of ±7% and found not to be influenced by cell density (0-30 g/L gel), cell type, and cell viability in gel beads. The oxygen diffusivity in the Ca-alginate gel beads was superior to that of the Ba-alginate gel beads, and the De in the Ca-alginate gel beads nearly equalled the molecular diffusion coefficient in the liquid containing the gel beads. The oxygen concentration profile in a single Ca-alginate gel bead was calculated and compared to the distribution of mycelia of Aspergillus awamori grown in that gel bead. This procedure indicated that the oxygen concentration profile is useful for the estimation of the thickness of the cell layer in a gel bead. Numerical investigation revealed that high effectiveness factors, greater than 0.8, could be obtained using microgel beads with a radius of 0.25 mm.
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  • 289
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    Biotechnology and Bioengineering 34 (1989), S. 1366-1373 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The immobilization of heparinase to tresyl-chloride-activated cellulose hollow fibers for the removal of heparin from the bloodstream was examined. Whole blood can be circulated through cellulose hollow fibers without hemolysis and the tresyl chloride chemistry provides a strong linkage which limits the release of the enzyme from the support. The tresylation and immobilization methods were modified and optimized to improve the heparinase activity retained by cellulose. Pretreatment of the hollow fibers with 0.05/V sodium hydroxide increased the degree of tresylation and the immobilization yield by a factor of five. The use of triethylamine as the organic base in the tresyl chloride activation resulted in threefold greater activity retention by the support than when pyridine was used. Together, sodium hydroxide pretreatment and triethylamine enhanced the activity retained by cellulose to 26.2 ± 7.0% of that bound to the support. The activity retention was also a function of the technique used for immobilization. The best results were achieved when the enzyme was applied to the activated fibers once every 12 to 24 h for a total of four times. The active enzyme loading on the fibers was 0.3 mg heparin degraded/h cm2 when 4.5 μg protein/cm2 was bound to the fibers.
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  • 290
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    Biotechnology and Bioengineering 33 (1989), S. 1191-1195 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Aspergillus niger JTS191 was capable of the conversion of β-ionone to a mixture of its derivatives that is utilized to an essential oil of tobacco. The authors attempted this microbial conversion in the presence of an organic solvent to improve its reaction rate. The addition of isooctane accelerated the microbial conversion of β-ionone. It took three days to complete the reaction whereas without isooctane it took more than six days. The addition of isooctane also improved the resistance of A. niger to the antifungal property of β-ionone. A. niger pellets were immobilized in hydrophobic polymer, PU-3, and applied for the microbial conversion of β-ionone. Further improvement of the resistance to the antifungal property of β-ionone was achieved by immobilization. PU-3 immobilized A. niger was repeatedly used for microbial conversion of β-ionone in the presence of isooctane for more than 480 hours.
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  • 291
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    Biotechnology and Bioengineering 33 (1989), S. 1267-1276 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Reversible competitive inhibitors of the three enzymes β-galactosidase, trypsin, and serum cholinesterase have been covalently attached to nonionic ethoxylated surfactants. The binding of the resulting affinity-derivatized surfactants to the respective enzymes has been quantified by measuring Michaelis-Menten inhibition constants with kinetic assays. The surfactant-inhibitor of serum cholinesterase, octaethylene glycol monohexadecy ether pyridinium (C16E8-PYR), was adsorbed in aqueous solution to an octadecyl-bonded reverse-phase silica packing in a 2 × 0.2 cm stainless steel test column. The ability of the test column to function as a high-performance affinity chromatography (HPAC) column was determined by applying a mixture of bovine serum albumin and cholinesterase (4:1 w/w). Virtually all of the cholinesterase bound and was eluted by applying a gradient in ionic strength. The applied cholinfesterase was recovered with a yield of over 90% and an 11-fold purification. An aliquot of raw horse serum was then purified in the same fashion with a yield of 84% and a 280-fold purification. The surfactant-inhibitor was easily removed from the column with an alcohol wash for sterilization, cleaning, or application of a different affinity ligand. Moreover, the ligand density on the column can be easily manipulated by adsorbing mixtures of derivatized and underivatized surfactants. Leakage of ligands from the support seems to be minimal since the cholinesterase affinity column was operated efficiently after being exposed to 24,000 column volumes of buffer. The application of this technique to high-capacity, high-throughput reversible affinity purifications is limited only by the ability to identify suitable ligands.
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  • 292
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    Biotechnology and Bioengineering 33 (1989), S. 1283-1289 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fermentation rates and intracellular compositions have been determined for alginate-entrapped Saccharomyces cerevisiae and for identical cells in suspension. Glucose uptake and ethanol and glycerol production are approximately two times faster in immobilized cells than in suspended cells. Phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy of fermenting immobilized and suspended cells shows differences in intermediate metabolite levels such as fructose-1,6 diphosphate, glucose-6-phosphate, and 3-phosphoglycerate and in internal pH. Carbon-13 NMR shows an increase in polysaccharide production. These data suggest that immobilization has accelerated the rate of glucose transport or of glucose phosphorylation. These effects of immobilization upon cell metabolism are observed in a very short period of time under conditions in which negligible DNA, RNA, or protein synthesis takes place.
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  • 293
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    Biotechnology and Bioengineering 34 (1989), S. 30-38 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The synthesis of 2-heptanone from octanoic acid by Caalginate/Eudragit RL entrapped spores of Penicillium roquefortii is performed in batch and continuous reactions. The measurement of the HCI solution needed to overcome the pH increase during the course of a reaction allows continuous monitoring of the reaction progress of batch as well as continuous processes without any aliquot sampling. The hydration of the biocatalyst prior to a bioconversion performance is of major importance to achieve reproducible results. A continuous reaction carried out at pH 6.5 for one month demonstrates the feasibility of such a process and shows that the reaction occurs with a Michaelian behavior, with KM = 1.82 mmol/L and rsm = 0.82 mmol/L h, and no intraparticle diffusion limitation is found. The complex behavior of the spores during batch reactions makes this kind of operation unsuitable for kinetic studies.
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  • 294
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    Biotechnology and Bioengineering 34 (1989), S. 18-29 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Acetone-butanol-ethanol (ABE) fermentation was successfully carried out in an immobilized cell trickle bed reactor. The reactor was composed of two serial columns packed with Clostridium acetobutylicum ATCC 824 entrapped on the surface of natural sponge segments at a cell loading in the range of 2.03-5.56 g dry cells/g sponge. The average cell loading was 3.58 g dry cells/g sponge. Batch experiments indicated that a critical pH above 4.2 is necessary for the initiation of cell growth. One of the media used during continuous experiments consisted of a salt mixture alone and the other a nutrient medium containing a salt mixture with yeast extract and peptone. Effluent pH was controlled by supplying various fractions of the two different types of media. A nutrient medium fraction above 0.6 was crucial for successful fermentation in a trickle bed reactor. The nutrient medium fraction is the ratio of the volume of the nutrient medium to the total volume of nutrient plus salt medium. Supplying nutrient medium to both columns continuously was an effective way to meet both pH and nutrient requirement. A 257-mL reactor could ferment 45 g/L glucose from an initial concentration of 60 g/L glucose at a rate of 70 mL/h. Butanol, acetone, and ethanol concentrations were 8.82, 5.22, and 1.45 g/L, respectively, with a butanol and total solvent yield of 19.4 and 34.1 wt %. Solvent productivity in an immobilized cell trickle bed reactor was 4.2 g/L h, which was 10 times higher than that obtained in a batch fermentation using free cells and 2.76 times higher than that of an immobilized CSTR. If the nutrient medium fraction was below 0.6 and the pH was below 4.2, the system degenerated. Oxygen also contributed to the system degeneration. Upon degeneration, glucose consumption and solvent yield decreased to 30.9 g/L and 23.0 wt %, respectively. The yield of total liquid product (40.0 wt %) and butanol selectivity (60.0 wt %) remained almost constant. Once the cells were degenerated, they could not be recovered.
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  • 295
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    Biotechnology and Bioengineering 34 (1989), S. 39-54 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Energetic and reaction-rate interactions between hydrogenic (hydrogen-producing) and hydrogenotrophic (hydrogen-consuming) bacteria were investigated in five perturbation experiments performed on steady-state, mixed-culture methanogenic CSTRs receiving ethanol, propionate, or both hydrogenic substrates. When a large quantity of propionate was suddenly added to a propionatefed CSTR, PH2 increased to 10-4 atm and propionate oxidation remained energetically favorable. When ethanol was added to a CSTR receiving ethanol, PH2 rose to 6.3 × 10-3 atm within 5 h. In both perturbations, PH2 remained at levels such that oxidation of the hydrogenic substrate remained energetically favorable throughout the transient. Sudden increase in ethanol concentration in the ethanol- and propionate-fed CSTR resulted in an increase in PH2 such that propionate oxidation became energetically unfavorable and was blocked. Propionate utilization resumed when the added ethanol was depleted and PH2 returned to its previous steady-state levels. Ethanol perturbation of ethanol- and propionate-fed CSTRs led to the formation of reduced products, including n-propanol and four-through seven-carbon n-carboxylic acids, when PH2 was elevated; these products disappeared after PH2 returned to previous, steady-state levels. The transformations were consistent with reaction energetics. Reduced product formation may have been a sink for reducing equivalents, as an alternative to oxidation for propionate utilization, as indicated by an electron equivalents balance over the time course of experiments.
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  • 296
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    Biotechnology and Bioengineering 34 (1989), S. 990-999 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The ability of algae and bacteria to accumulate heavy metals from the surrounding environment is a widely recognized phenomenon that has a number of important implications. This work reports on the development of a quantitative model that addresses the basic mechanisms inherent in many uptake processes. The model postulates two mechanisms: an initial rapid metal ion uptake due to attachment onto the cell wall followed by a relatively slow uptake due to membrane transport of the metal into the cell. The mathematical model has been tested using the alga Chlorella vulgaris in the presence of cadmium and zinc in solution under various experimental conditions.
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    Biotechnology and Bioengineering 34 (1989), S. 971-989 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Ramkrishna and his co-workers have developed so-called cybernetic models which purport to describe, among other things, how microorganisms make choices when presented with two or more functionally equivalent, or substitutable, nutrients that are sources of carbon and available energy. In general, however, organisms are presented with choices not just between nutrients that are substitutable for one another, but also between sets of nutrients some of which are by no means substitutable for one another. It is postulated herein that the main ideas of cybernetic modeling apply to these more general choices as they seem to apply to the choices considered by Ramkrishna and his co-workers. Some consequences of the postulate are worked out for steady-state growth situations where two, or in one case three, nutrients limit or potentially limit growth rate. If predicted phenomena are observed experimentally so as to verify the postulate, a significantly improved basis for understanding growth of microorganisms in practical fermentation media as well as in natural situations will be provided by this application of cybernetic modeling ideas.
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    Biotechnology and Bioengineering 34 (1989), S. 1037-1044 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Gramicidin S synthetase, the enzyme complex catalyzing the biosynthesis of the antibiotic gramicidin S in Bacillus brevis, is subject to O2-dependent in vivo inactivation during exponential aerobic growth after reaching a peak in specific activity. The five amino acid substrates of the synthetase are capable of stabilizing its activity to varying degrees in whole cells shaken aerobically. Depending on the time of cell harvesting before, during, or after the peak in intracellular gramicidin S synthetase specific activity, the enzyme has a long, medium, or short half-life, respectively. The kinetic profiles of gramicidin S synthetase in B. brevis cells indicate that both the kinetics of synthetase loss and the degree of its amino-acid-mediated stabilization are a strong function of the cells' physiological development.
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  • 299
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    Biotechnology and Bioengineering 34 (1989), S. 1331-1336 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 2 Ill.
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  • 300
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    Biotechnology and Bioengineering 34 (1989), S. 1341-1356 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Microbial desulfurization of coal by pyrite oxidizing bacterial enrichment cultures has been studied in air-agitated slurry reactors of 4- and 20-L volumes. Batch experiments showed that inoculation with an active bacterial culture is essential to minimize the lag phase, although a considerable number of pyrite oxidizing bacteria was found on the coal prior to desulfurization. For detailed investigations of kinetics, energy requirements, and technical applicability, a bioreactor equipment consisting of a cascade of eight stages was developed and operated continuously. Microbial desulfurization of coal - monitored by measuring the axial profile of dissolved iron concentration, real and maximum oxygen consumption rates, and cell concentration - at pulp densities to 30% was performed over a period of 200 days without any disturbances concerning the aeration system, fluidization, transport of solids and microbial growth. At a pulp density of 20%, a pyrite conversion of 68% was achieved after the third reactor stage at a total residence time of five days in the first three stages. The kinetics of pyrite degradation were found to be well described by a rate equation of first order in pyrite surface area concentration if the pyrite is directly accessible for microbial attack. Rate constants were determined to 0.48 mg pyrite/(cm2 day) in the first and to 0.24 mg pyrite/(cm2 day) in the following reactor stages. Kinetic models taking into account adsorption/desorption as well as growth kinetics failed to describe the observed reaction rates. However, a model treating pyrite degradation and microbial growth kinetics formalistically seems to be applicable when backmixing between the reactor stages can be avoided. The advantage of a multistage reactor in comparison to single-stage equipment was shown by calculation. To obtain a pyrite conversion of 68%, a three-stage reactor would require only 58% of the volume of single-stage equipment.Measurement of oxygen consumption rates proved to provide quickly and easily measurable parameters to observe microbial coal desulfurization in technical scale: the real oxygen consumption rate is correlated to the pyrite oxidation rate and the maximum oxygen consumption rate is correlated to the concentration of viable cells. The Yo/s coefficient for the amount of oxygen consumed per mass unit of pyrite oxygen was determined to approximately 0.33 in comparison to 1.0 which can be calculated from stoichiornetry. This could yet not be explained. Chemical leaching experiments as well as sulfur analyses of desulfurized coal samples showed that the microorganisms play the main role in degradation of pyrite from coal and that pyrite oxidation by ferric iron can be neglected.
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