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  • Biochemistry and Biotechnology  (589)
  • 1985-1989  (589)
  • 1950-1954
  • 1988  (589)
  • 1
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 31 (1988), S. 160-167 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Six endoglucanases (Endo I, II, III, IV, V, and VI), three exoglucanases (Exo I, II, and III), and a β-glucosidase (β-gluc I) isolated from a commercial cellulase preparation of Trichoderma viride origin were examined as to their activities on xylan ex oat spelts. Endo I, II, and III as well as Exo II and III showed no activity toward xylan and were classified as specific glucanases. Less specificity was found for the endoglucanases Endo IV, V, and VI, Exo I, and β-gluc I, whose enzymes were able to hydrolyze xylan. With respect to product formation these xylanolytic cellulases fit the classification of xylanases generally accepted in the literature. Kinetic experiment with xylan, CM-cellulose, and p-nitrophenyl-β-D-glucoside revealed that Endo IV, V, an VI and Exo I prefer to hydrolyze β-1, 4-D-glucosidic linkages. β-Gluc I showed no clear substrate preference.
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  • 2
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    Biotechnology and Bioengineering 31 (1988), S. 173-178 
    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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  • 3
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    Biotechnology and Bioengineering 31 (1988), S. 179-182 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 4
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    Biotechnology and Bioengineering 31 (1988), S. 895-904 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Native starch granules from wheat have been subjected to enzymatic depolymerization with an α-amylase from Bacillus subtilis. Crystallites made from short-chain amylose and residues from mild acid hydrolysis have been also tested. Electron microscopy, particle size analysis, DSC, and x-ray diffractometry reveal that enzymatic degradation occurs granule by granule. Gel permeation chromatography shows off the macromolecular nature of the remaining material. In contrast, acid erodes simultaneously all the granules, leading to a splitting into small particles. Crystalline fractions are completely degraded by α-amylase. These results support evidence for an active disentanglement of chains, carried out by the different subsites of α-amylase molecules. A simple mathematical treatment is proposed to explain the results of the kinetics.
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  • 5
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    Biotechnology and Bioengineering 31 (1988), S. 958-968 
    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 operational, parameters, such as hydraulic retention time, organic loading rate, influent substrate concentration, pH, and temperature, on the performance of the first phase of anaerobic digestion has been investigated. A complex substrate based on beef extract was used, and six series of experimental runs were conducted, each one showing the effect of one operational variable. The predominant fermentation products were always acetic and propionic acid, independent of the values of the operational parameters. For initial COD concentrations and hydraulic retention times above the critical values identified as 3 g/L and 6 h, respectively, the degree of acidification achieved was between 30 and 60%. The degree of acidification was found to increase with the hydraulic retention time and decrease with the influent substrate concentration and organic loading rate, while the opposite held true for the rate of product formation. Furthermore, it has been demonstrated that acidification is primarily determined by the hydraulic retention time and the rate of product formation by the influent substrate concentration. The concentration of the acetic acid produced was found to depend on the operational parameters. However, the concentration of propionic acid produced depended only on the substrate availability with a consistent proportion of 8% initial COD converted to it. The optimum pH and temperature were 7 and 40°C, respectively. The percentage of acetic acid as a proportion of the total volatile fatty acids produced was found to increase with increasing pH and temperature, while the percentage of propionic acid seemed to decrease accordingly. Finally the effect of the temperature on the rate of acidification followed an Arrhenius type equation with an activation energy equal to 4739 cal/mol.
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  • 6
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    Biotechnology and Bioengineering 31 (1988), S. 979-983 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new method for determination of the binodial of an aqueous two-phase system, using spectrophotometric measurements of the turbidity, is described in this article. The method is especially designed to characterize phase systems composed of polydisperse phase components. It gives information about the area representing the transition from homogeneous solution to a two-phase system. The two-phase systems studied were first a conventional Dextran T40-polyethylene glycol 20M (PEG 20M) system, then a less-well-defined phase system based on PEG 20M and partially hydrolyzed starch. The PEG 20M-starch system could be changed with respect to the volume ratio between the phases with time using hydrolytic enzymes, and the possibility of using the glucose released from the starch polymer is pointed out. Then the system is transformed to an extractive biconversion where the bottom phase polymer also served as the substrate.
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  • 7
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    Biotechnology and Bioengineering 31 (1988), S. 984-994 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The use of microporous membranes has been examined for the recovery of precipitated protein suspensions and related soluble protein. Membrane flux rates and soluble protein transmissions are reported for unstirred batch-cell studies and cross-flow experiments. The unstirred batch-cell gave soluble protein transmissions in the range 80-100% for feeds containing either soluble protein or a mix of soluble and isoelectrically precipitated protein. In all cases a sharp decline in flux was observed which was, for example, considerably greater for soluble protein at its isoelectric point, pH 4.6, than at pH 8.8. The presence of precipitated protein led to a further decrease in flux rate. In cross-flow studies, flux decline was eventually accompanied by a significant decline in soluble protein transmission. The flux protein-transmission characteristics of microporous membranes are discussed in terms of the interaction of the soluble and precipitated protein with the membrane.
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  • 8
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    Biotechnology and Bioengineering 31 (1988), S. 995-1005 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A membrane-covered oxygen electrode was used to measure oxygen diffusion coefficients and solubilities in aqueous glucose solutions and various fermentation media following a newly developed methodology. The fermentation media studied were tryptic soy broth and those for fermentations of Penicillium chrysogenum, Saccharomyces cerevisiae, and Micrococcus glutamicus. The experimental results of oxygen diffusion coefficients and solubilities in glucose solutions were in good accord with the literature data. As for the fermentation media, both oxygen diffusion coefficients and solubilities were found to decrease with an increased fractional composition of these media, and log-additive behaviors of the oxygen diffusion coefficients and solubilities in fermentation media were observed.
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  • 9
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    Biotechnology and Bioengineering 31 (1988), S. 1006-1009 
    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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  • 10
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    Biotechnology and Bioengineering 31 (1988), S. 1010-1011 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 11
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    Biotechnology and Bioengineering 32 (1988) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 12
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    Biotechnology and Bioengineering 32 (1988), S. iii 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 13
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    Biotechnology and Bioengineering 31 (1988), S. 495-501 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 14
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    Biotechnology and Bioengineering 31 (1988), S. 487-494 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Gas holdup and oxygen transfer studies in non-Newtonian suspensions of cellulose fibres conducted in two large (0.098 m3 each) reactors are described. Both reactors - a bubble column and a similar internal loop airlift - were unusual in that they had rectangular cross-sections. In all cases gas holdups and kLaL declined with increasing solid concentration and, under identical conditions, the bubble column performed better than the airlift. The fluid systems used were carefully selected to represent mould fermentation broths.The behavior of true mass transfer coeffcient kL with changes in bubble size is discussed for these systems.
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  • 15
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    Biotechnology and Bioengineering 31 (1988), S. 502-506 
    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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  • 16
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    Biotechnology and Bioengineering 31 (1988), S. 287-294 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simple adaptive control algorithm, for which theoretical stability and convergence properties had been previously demonstrated, has been successfully implemented on a biomethanation pilot reactor. The methane digester, operated in the CSTR mode was submitted to a shock load, and successfully computer controlled during the subsequent transitory state.
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  • 17
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    Biotechnology and Bioengineering 31 (1988), S. 295-299 
    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 temperature, pH, and substrate and product concentrations on the oxidation rate of ferrous iron by biofilm of Thiobacillus ferrooxidans was determined. The experiments were performed in an inverse fluidized-bed biofilm reactor in which the biofilm thickness was kept constant at 80 μm. Oxygen concentration and diffusion through the biofilm did not limit the oxidation rate. The oxidation rate was almost unaffected by temperature between 13 and 38°C, pH between 1.3 and 2.2, ferric iron concentration up to 14 g/L, or ferrous iron concentration from 4 to 13 g/L. The kinetics of the process was described by the Monod equation with respect to the mass of the biofilm and with ferrous ions as the limiting substrate.
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  • 18
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    Biotechnology and Bioengineering 31 (1988), S. 304-310 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A segregated model of multicopy plasmid propagation has been formulated which incorporates plasmid replication and partition functions, as well as the effect of plasmid presence on host growth rate. Growth of plasmid-free cells in selective medium is explicitly analyzed. The model parameters can be determined from experimentally measurable quantities. Propagation of a recombinant multicopy plasmid in the yeast Saccharomyces cerevisiae is analyzed using this model.
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  • 19
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    Biotechnology and Bioengineering 31 (1988), S. 300-303 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A total of 58 yeast strains from 12 genera were assayed for their ability to grow and ferment carbohydrates in standard Durham tube test at 40, 43, and 46°C. Based on the kinetic parameters for glucose fermentation in shaken flask cultures, the strain Fabospora fragilis CCY51-1-1 was chosen for further studies. It reached about 56.0 and 35.0 g ethanol/L from ∼140 g glucose/L at 43 and 46°C in less than 48 h, respectively. Trichoderma reesei cellulase preparation (400 FPU/L) had not distinct effect on the ethanol yield and biomass production by the selected strain in the first 12 h fermentation at 46°C. Later a negligible decrease in both yields was observed. It was found that Fabospora fragilis did not grow or produce ethanol at 46°C as tho initial ethanol concentration overcame 40 g/L.
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  • 20
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    Biotechnology and Bioengineering 31 (1988), S. 311-320 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The objective of this work was to relate macroscopically measurable on-line fermentation parameters such as dissolved oxygen, off-gas oxygen and carbon dioxide, and cell mass, to the controlled production of key intracellular enzymes under carbon limited conditions. Both batch and perturbed batch aerobic fermentations were performed using two different strains of Escherichia coli, with glucose and lactose as the sole carbon sources. The two strains differed from each other only in the lac operon region of their genome. The parent strain, E. coli 3000, was inducible for the enzyme β-galactosidase. The other strain, E. coli 3300, was a constitutive mutant in the production of β-galactosidase. In all experiments, off-line assays of sugars and β-galactosidase activity were performed. It was observed that there is a clear relationship between the macroscopic on-line measurements, dissolved oxygen tension, carbon dioxide evolution rate and oxygen uptake rate, and the microscopic control phenomena of catabolite repression, catabolite inhibition, and inducer repression.
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  • 21
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    Biotechnology and Bioengineering 32 (1988), S. 64-67 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Photoacoustic spectroscopy was used to monitor photo synthetic electron transfer in native and immobilized thylakoid membranes. The photoacoustic parameter φr′ (the percentage of absorbed energy that is stored in photo chemical intermediates) and i50 (the half-saturation modulated light intensity) were directly correlated to electron transfer rates. As previously shown, thylakoids immobilized in an albumin-glutaraldehyde matrix were more resistant to aging. The inhibitory effects of the immobilization procedure and of aging at 4°C were detected as a decrease in i50 values. In analogy with enzyme kinetic analysis, the effect could be characterized as a competitive type of inhibition. Photoacoustic measurements are performed in conditions similar to a working bioreactor cell with regards to the sample preparation.
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  • 22
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    Biotechnology and Bioengineering 31 (1988), S. 559-566 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Naphthalene-2-sulfonate was degraded by submerse growing Pseudomonads in a chemostat culture. The kinetic parameters for the Monod equation, including Pirts maintenance energy, were calculated from these experiments regarding naphthalene-2-sulfonate as substrate and oxygene as cosubstrate. By immobilizing the bacteria on sand particles, the degradation of naphthalene-2-sulfonate was carried out in a specialy designed three-phase airlift-loop reactor in a completely fluidized state. From these experiments, the influence of biofilm diffusion limitation on reaction kinetics and criteria for stable biofilm formation on sand particles were obtained.
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  • 23
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    Biotechnology and Bioengineering 31 (1988), S. 553-558 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A reactor, using the enzymatic electrocatalysis scheme, was developed on a laboratory preparative scale for the catalytic oxidation of glucose into gluconic acid. Glucose oxidase was directly immobilized on the surface of a carbon felt electrode and a solution of glucose and benzo-quinone passed through the electrode in order to regenerate the electron acceptor. The reactor was able to produce continuously 1.5 g gluconate/h with a catalytic current of 0.4 A. This gave a high productivity ca. 100 g/h/L reactor. A one-dimensional model was developed which demonstrated the efficiency of coupling between enzymatic and electrochemical reactions due to the proximity of the two reaction sites. For example the catalytic current was practically independent of mass transfer parameters. The operational stability of immobilized glucose oxidase was increased 50 times at least when electroregenerated benzoquinone was used as oxidant instead of oxygen.
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  • 24
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    Biotechnology and Bioengineering 31 (1988), S. 567-578 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In production-scale bioreactors microorganisms are exposed to a continually changing environment. This may cause loss of viability, reduction of the yield of biomass or desired metabolites, and an increase in the formation of by-products. In fed-batch production of baker's yeast, profiles may occur in substrate and oxygen concentrations and in pH. This article deals with the influence of a periodically changing oxygen concentration on the growth of baker's yeast in a continuous culture. Also, influences on the production of ethanol, glycerol, acetic acid, and on the composition of the cells were investigated. It was found that relatively fast fluctuations between oxygen-unlimited and oxygen-limited conditions with a frequency of 1 or 2 min had a distinct influence on the biomass and metabolite production. However, RNA, protein, and carbohydrate contents measured in cells exposed to fluctuations differed little from those in cells from an oxygen-unlimited or an oxygen-limited culture. The respiration and fermentation capacities of cells exposed to fluctuations can be larger than the capacities of cells grown under oxygen-unlimited conditions.
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  • 25
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    Biotechnology and Bioengineering 32 (1988), S. 348-355 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Development of a novel two-layer anaerobic biofilm model is based on substrate utilization kinetics and mass transport. The model is applied to steady-state conditions for a fixed-film anaerobic reactor. The microbial film is considered to consist of two distinct biofilm layers, one adjacent to the second, with an acidogenic bacteria biofilm forming the outer layer and a methanogenic film the inner one. The model assumes that sugars are only metabolized by the first layer and converted into volatile fatty acids (VFA), while fatty acids are taken up only by the inner layer. The model is able to predict both substrate flux net uptake and methane production for steady-state conditions. The results of modeling agree with methane production experimental data published elsewhere. Further, the model shows why layered fixed-film reactors can withstand high and inhibitory concentrations of volatile fatty acids as well as severe overloading without failure.
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  • 26
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    Biotechnology and Bioengineering 32 (1988), S. 356-362 
    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 Streptomyces griseus for the production of cycloheximide is similar to other antibiotic fermentations in that product synthesis is subject to feedback regulation and the desired product is degraded in the fermentation broth. The productivity of this fermentation can thus be dramatically increased by removing the antibiotic from the whole broth as it is produced. One means for effecting this on-line product removal is to contact the whole fermentation broth with neutral polymeric resin immobilized in hydrogel beads. The antibiotic adsorbs to the immobilized resin via hydrophobic interactions. In this work, the adsorption of the antibiotic onto the immobilized resin was characterized. A biochemical model of the fermentation was then used to describe the time profiles of biomass, substrate, and antibiotic in a fermentation system in which whole broth is circulated from the fermentor through a continuously stirred extractor containing the adsorbent beads. Various operating conditions were examined to optimize the productivity of the fermentation.
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  • 27
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    Biotechnology and Bioengineering 32 (1988), S. 669-676 
    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 production process of maltotetraose was investigated by using immobilized maltotetraose (G4)- forming amylase (1,4-α-D-glucan maltotetraohydrolase, EC3.2.1.60) from Pseudomonas stutzeri adsorbed on a macroporous hydrophobic resin. The maximum reaction rate was obtained at 55°C and the activation energy of hydrolysis by immobilized G4-forming amylase was calculated to be 8.45 kcal/mol. The maltotetraose yield was greatly influenced by the flow rate of substrate solution, its concentration, and the immobilized enzyme activity. The newly defined factor “specific space velocity” was successfully introduced to normalize the operating parameters. Using this factor, the immobilized enzyme reactor then can be simulated and the operating dynamics can be determined.
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    Biotechnology and Bioengineering 32 (1988), S. 664-668 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Invertase was ionically immobilized on the poly(ethylene-co-vinyl alcohol) hollow fiber inside surface, which was aminoacetalized with 2-dimethylaminoacetaldehyde dimethyl acetal. Immobilization and enzyme reaction were carried out by letting the respective solutions pass or circulate through the inside of the hollow fiber, and the activity of invertase was determined by the amount of glucose produced enzymatically from sucrose. Immobilization conditions were examined with respect to the enzyme concentration and to the time, and consequently the preferable conditions at room temperature were found to be 5 μg/mL of enzyme concentration and 4 h of immobilization time. Under those conditions the immobilization yield and the ratio of the activity of the immobilized invertase to that of the native one were 89 and 80%, respectively. For both repeating and continuous usages, the activity fell to ca. 60% of the initial activity in the early stage and after that almost kept that value. The apparent Michaelis constant Km′ for the immobilized invertase decreased with increasing the flow rate of the substrate solution, to be close to the value for the native one. Furthermore, the possibility of the separation of the enzymatically formed glucose from the reaction mixture through the hollow fiber membrane was preliminarily examined.
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    Biotechnology and Bioengineering 32 (1988), S. 396-399 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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    Biotechnology and Bioengineering 32 (1988), S. 174-183 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Batch foam separation has been employed to separate Saccharomyces carlsbergensis cells from their broth without the use of any external surface-active agent. A model has been developed to predict the foamate cell concentration as well as the variation of cell concentration in the bulk liquid in the foam column as a function of time. The model assumes a linear equilibrium relation between the cell concentrations at the interface and the bulk. The foam has interface as well as interstitial liquid. The interface is assumed to be in equilibrium with the interstitial liquid, which in turn is assumed to have the same concentration as the bulk. The interfacial area is calculated by assuming the foam bubbles to be pentagonal dodecahedral in shape. The model has been found to explain the results of foam separation of cells quite well, particularly with respect to the effect of bubble size and aeration rate.
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    Biotechnology and Bioengineering 32 (1988), S. 159-173 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Anaerobic degradation performance of a laboratory-scale packed-bed reactor (PBR) was compared with two fluidized-bed biofilm reactors (FBRs) on molasses and whey feeds. The reactors were operated under constant pH (7) and temperature (35°C) conditions and were well mixed with high recirculation rates. The measured variables were chemical oxygen demand (COD), individual organic acids, gas composition, and gas rates. As carrier, sand of 0.3-0.5 mm diameter was used in the FBR, and porous clay spheres of 6 mm diameter were used in the PBR. Startup of the PBR was achieved with 1-5 day residence times. Start-up of the FBR was only successful if liquid residence times were held low at 2-3 h. COD degradations of 86% with molasses (90% was biodegradable) were reached in both the FBR and PBR at 6 h residence time and loadings of 10 g COD/L day. At higher loadings the FBR gave the best performance; even at 40-45 g COD/L day, with 6 h residence times, 70% COD was degraded. The PBR could not be operated above 20 g COD/L day without clogging. A comparison of the reaction rates show that the PBR and FBR per formed similarly at low concentrations in the reactors up to 1 g COD/L, while above 3 g COD/L the rates were 17.4 g COD/L day for the PBR and 38.4 g COD/L day for the FBR. This difference is probably due to diffusion limitations and a less active biomass content of the PBR compared with the fluidized bed.The results of dynamic step change experiments, in which residence times and feed concentrations were changed hanged at constant loading, demonstrated the rapid response of the reactors. Thus, the response times for an increase in gas rate or an increase in organic acids due to an increase in feed concentration were less than 1 day and could be explained by substrate limitation. Other slower responses were observed in which the reactor culture adapted over periods of 5-10 days; these were apparently growth related. An increase in loading of over 100% always resulted in large increases inorganic acids, especially acetic and propionic, as well as large increases in the CO2 gas content. In general, the CO2 content of the gas was very low, due to the large amount of dissolved CO2 that exited with the liquid phase at low residence times. The performance of the FBR with whey was comparable to its performance with molasses, and switching of molasses to whey feed resulted in immediate good performance without adaptation.
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  • 32
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    Biotechnology and Bioengineering 32 (1988), S. 184-191 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new combined bioreactor-separator system was designed and its operational feasibility demonstrated in order to develop a bioprocess that enables us to handle simultaneous biotransformation and recovery of product by crystallization. Enzymatic conversion of L-aspartate to L-alanine by L-aspartate β-decarboxylase from Pseudomonas dacunhae (ATCC 21192) was used as a model system for this study to demonstrate the principles involved in the bioprocess design. Immobilized cells of P. dacunhae containing the enzyme were fluidized in a tapered column type of bioreactor and a filter-crystallizer combination was used as a separator unit in our experimental system.It was found that almost a theoretical yield was achieved, and the process control for both the bioreactor operation and separation was relatively easy. The Production systems, namely, the recirculating bioreactor separator combination system and the conventional batch reactor system, were analyzed and compared based on the results obtained form this study, and it was found that a significant cost reduction, by about 20%, can be achieved when the recirculating bioreactor-separator combination system was employed. Based on these findings, it is anticipated that the conceptual design of the bioreactor-separator combination system evaluated in this study has some potential for industrial application.
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  • 33
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    Biotechnology and Bioengineering 32 (1988), S. 192-204 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The performance of differential contactors for use in extractive fermentation is complicated by the effects of product formation in the contactor. When product formation is significant, approximate analytical solutions are presented for the performance of the contactor for two limiting cases: high and low substrate concentrations. When products are formed at a constant rate, there is a minimum raffinate solute concentration that can be obtained, in contrast to the behavior of a column in the absence of product formation. General equations describing the behavior of the system for product formation with backmixing in both phases are presented. The case of a stripping factor not equal to unity is considered.
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  • 34
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    Biotechnology and Bioengineering 32 (1988), S. 966-974 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Nuclear magnetic resonance (NMR) spectroscopic analysis of whole cells is an important emerging technique for noninvasive and nondestructive monitoring of cell physiology. However, this technique requires extremely high cell densities. Attempts to maintain densities above the carrying capacity of a maintenance system result in the demise of the entire culture. To define conditions for maintaining mammalian cells at high densities for NMR studies, we have designed a bioreactor to operate under defined, oxygen-limited conditions within an NMR spectrometer. The bioreactor utilizes hollow fibers to deliver nutrients and remove wastes from an agitated cell suspension. The mass transfer properties of the fibers with respect to oxygen were determined. Ehrlich Ascites Tumor (EAT) cells were supplied with glutamine as the respiratory carbon source. The maximum viable cell density supported by a given oxygen concentration in the fluid flowing through the fiber lumen was predicted and then confirmed experimentally on the bench and in the spectrometer.
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    Biotechnology and Bioengineering 32 (1988), S. 975-982 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Results are presented which show how the microcarrier concentration affects the hydrodynamic environment in animal cell bioreactors. At low levels of agitation, no physical effects of microcarrier concentration were found. However, cell growth was strongly influenced by cell concentration. At high levels of agitation, a strong detrimental effect of microcarrier concentration was found. A new mechanism of hydrodynamic damage was identified which is second order in microcarrier concentration. The identification of this mechanism adds to the fundamental understanding of hydrodynamic phenomena in microcarrier bioreactors.
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  • 36
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    Biotechnology and Bioengineering 31 (1988), S. 675-681 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel human protein exhibiting erythroid differentiation activity was discovered in the culture fluids of phorbol ester-stimulated human cells. The differentiation assay system involving Friend virus-derived mouse leukemia cells was used. THP-1 cells of myelomonocytic origin were typical producers. 4β-Phorbol 12-myristate 13-acetate (PMA) was essential for inducible excretion of the erythroid differentiation factor (EDF). The factor was stable toward heat and pH (acidic or alkaline) but lost its activity on pronase treatment, which suggested its proteinous nature. After an optimization of the condition, production of EDF was performed on a 200-L scale for purification of the protein.
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    Biotechnology and Bioengineering 31 (1988), S. 682-695 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A mathematical model was developed to simulate the operation of an activated sludge system with biomass support particles. Based on the results of this simulation study, it would appear that a system with biomass particle supports offers several advantages not offered by conventional systems. These include the ability to resist failure from large hydraulic surges; stable nitrification under transient inhibitory conditions resulting from temperature changes, hydraulic surges, and/or toxic chemicals; and the ability to establish stable operating conditions with respect to both carbon oxidation and nitrification at short hydraulic retention times and low sludge ages.
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  • 38
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    Biotechnology and Bioengineering 32 (1988), S. 983-992 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A high surface area hollow fiber reactor was developed for mammalian cell culture. The reactor employs an interfiber gel matrix of agar or collagen for cell support. A model was developed to predict cell density as a function of fiber spacing. Optimum spacings are calculated for two sizes of Celgard hollow fibers. Ehrlich Ascites Tumor (EAT) cells were grown to an estimated density of 1.1 × 108 viable cells/mL in the extracapillary space - corresponding to an overall reactor density of 7 × 107 cells/mL. On the basis of available kinetic and diffusivity data, the model predicts that lactate accumulation may limit cell growth in the early stage of medium utilization, while oxygen delivery becomes limiting at later stages.
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  • 39
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    Biotechnology and Bioengineering 32 (1988), S. 993-1000 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Monoclonal antibodies are being manufactured for clinical trials in suspension culture at the 1300-L scale. Suspension culture offers some advantages relative to high-density mammalian cell culture methods; in particular, the ability to closely monitor the behavior of cells in a homogeneous environment. Computer control and on-line mass spectrography of exit gases provide instantaneous information about the culture metabolic activity. Air sparging and agitation by marine impeller provide aeration sufficient to maintain a constant dissolved oxygen tension at cell concentrations up to 5.0 × 106 cells/mL without causing apparent cell damage.
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    Biotechnology and Bioengineering 32 (1988), S. 803-812 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Substrate and energy costs of the production of exocellular enzymes from glucose and citrate by B. Iicheniformis S1684 as well as molar growth yields corrected for these costs of product formation were calculated using data from chemostat experiments. The calculations showed that 1.46-1.73 mol glucose and 2.31-2.77 mol citrate are needed for formation and excretion of 1 mol protein. Consequently, the values of the maximal product yield from substrate (Ypsm' g/mol) are 80 〈 Ypsm 〈 95 when product is formed from glucose and 50 〈 Ypsm 〈 60 when product is formed from citrate. The higher substrate costs for product formation from citrate are due to a higher level of CO2 production during protein formation and a higher substrate requirement for the energy supply of product formation and excretion than when product is formed from glucose. The theoretical ATP requirement for protein synthesis could be determined reasonably well, but the energy costs of protein excretion could not be determined exactly. The energy costs of protein formation are higher than those of biomass formation or protein excretion. Molar growth yields corrected for the substrate costs of product formation were high, indicating a high efficiency of growth.Growth and production parameters were determined as well from experimental data of recycling fermentor experiments using a parameter optimization procedure based on a mathematical model describing biomass growth as a linear function of the substrate consumption rate and the rate of product formation as a linear function of biomass growth rate. The fitting procedure yielded two growth and production domains during glucose limitation. In the first domain the values for the maximal growth yield and maintenance coefficient were in agreement with those found in chemostat experiments at corresponding values of Yspm. Domain 2 could be described best with linear growth and product formation. In domain 2 the rate of product formation decreased and more substrate became available for biomass formation. As a consequence the specific growth rate increased in the shift from domain 1 to 2. Domain 2 behavior most probably is caused by the rel-status of B. Iicheniformis S1684.
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    Biotechnology and Bioengineering 32 (1988), S. 826-830 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 32 (1988), S. 831-834 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 32 (1988), S. 835-840 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 44
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    Biotechnology and Bioengineering 32 (1988), S. 263-270 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A laboratory model constant-depth film fermentor was developed. Film grew on the surface of polytetrafluoroethylene (PTFE) plugs and was limited to a predetermined depth by mechanically removing excess film. Six film-forming organisms were isolated from river water and used to assess the operating characteristics of the fermentor. Film accumulation was logarithmic, and a steady state was maintained. Electron micrographs show early film development. The fermentor enables film to be grown on any substratum and allows discrete, reproducible, and representative samples to be taken.
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  • 45
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    Biotechnology and Bioengineering 32 (1988), S. 277-288 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this study, a cephalosporin C producing strain, Cephalosporium acremonium (ATCC 36225), was chosen to determine the optimal conditions that maximize antibiotic production in a mixed substrate of glucose and sucrose. A model for cell growth and cephalosporin C production at different pH and temperature was developed and the associated parameters were evaluated experimentally. Pontryagin's maximum principle, in conjunction with the model, was used to predict the optimal temperature and pH control profiles to maximize the production of antibiotic.
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  • 46
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    Biotechnology and Bioengineering 32 (1988), S. 271-276 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A kinetic model of ethanol fermentation conducted under a variety of conditions in a continuous four-stage reactor is proposed. The expressions for specific growth and product formation rates are: \documentclass{article}\pagestyle{empty}\begin{document}$$ \mu = \mu _0 {\rm exp( - }k_1 P){\rm (1 - }X/X_1) \\ \nu _P = \nu _0 {\rm exp( - }k_2 P){\rm (1 - }X/X_2) \\ $$\end{document} Parameters were identified by nonlinear programming and shown to fit data correctly for steady states of seven different experiments. The product inhibition constants were of 27 and 84 g/L, respectively. Secondary inhibitions were represented by the linear biomass term. The proposed model gave a better description of phenomena than one which only took ethanol inhibition into account. The same model also fitted batch fermentation data, with only some parameters altering significantly. The use of this model for on-line purposes is discussed.
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  • 47
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    Biotechnology and Bioengineering 32 (1988), S. 289-294 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Poly(styrene) microcapsules, prepared by depositing the polymer around emulsified aqueous droplets, were coated with a synthesized polyelectrolyte; i.e., copolymer of maleic acid (MA) with methyl vinyl ether (MVE), co-poly(MA, MVE), or with styrene (St), copoly(Ma, St). The permeability of the capsule membrane was investigated under various pHs of the outer medium using n-propyl alcohol as a permeant. It became apparent that either copoly(MA, St)- or copoly(MA, MVE)-coated microcapsules function as a pH-sensitive capsule. In particular, the former showed a dramatic change of the permeability in response to small differences in pH (5-6). By reference to the viscometric and electrophoretic studies of both copolymers, these were interpreted as being due to a pH-induced alteration of the configuration of the copolymer coating on the surface of the capsule membrane. When sucrose was hydrolyzed in an aqueous suspension of the copoly(MA, St)-coated capsules into which invertase was loaded, the hydrolytic reaction was initiated at pH 5. 5 and stopped at pH 4. 5. Such initiation-cessation control was repeated reversibly without damaging the capsules.
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  • 48
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    Biotechnology and Bioengineering 31 (1988), S. 770-782 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Batch fermentations of glucose to ethanol by Z. Mobilis.(ATCC 10988) were examined in several semidefined nutrient media. The measurement of acid produced by the microorganism was used to study its transient fermentation characteristics. Limitation of nitrogen source in the semidefined medium of Rogers and coworkers2 was found to limit the growth of this microorganism in the late stages of batch fermentations, when the initial glucose concentration was 75 g/L and higher. The microorganism exhibits a preference for inorganic nitrogen over preformed organic nitrogen provided by yeast extract. The microbial growth occurs exponentially in the presence of ammonium sulfate and yeast extract. However, in the absence of ammonium sulfate, the growth occurs in a linear fashion. The “linear” growth phase is characterized by poor cell-mass yields, and during this phase, growth and ethanol production are decoupled. An improved semi-defined growth medium is established which supports better growth rate and cellular yield, without affecting the ethanol yield.
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    Biotechnology and Bioengineering 32 (1988), S. 910-915 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 32 (1988), S. 916-919 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 32 (1988), S. 920-922 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 32 (1988), S. 1128-1137 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The interrelationships between the three parts of the air lift reactor, the riser, the downcomer, and gas-liquid separator, were examined with relation to the overall mass transfer in the reactor. This involved studying the mass transfer of oxygen from the gas phase to the liquid phase for 20 different reactor geometries. Both one- and two-sparger systems were studied. It was demonstrated that the gas-liquid separator plays a major role in reactor behavior and must be considered in reactor design. It was found that the overall reactor mass transfer coefficient KLA could be correlated to the pneumatic power of gas input per total dispersion volume (P/VD) and to the true riser superficial gas velocity JGR for all experimental conditions examined. The KLA is directly related to the P/VD with an exponent of approximately 1. “Two-sparger” systems, where an auxiliary gas sparger is placed near the downcomer entrance, have higher ab solute values for KLA than single-sparger systems.
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  • 53
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Notes: An activated sludge conditioned to a low concentration of phenol was used for the biodegradation of phenol in the presence of Chlorophenols, measuring the inhibiting action of all the latter compounds and expressing it as p/50. As a structural property to be correlated with the measured values, the lipophilicity (log P) of the Chlorophenols was reevaluated with a new apparatus. Values of p/50p are better correlated with the number of chloro substituents in the inhibitor, provided a specific effect of substituents ortho to the phenolic function is considered. New results are compared with previous measurements effected on a different activated sludge.
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    Biotechnology and Bioengineering 31 (1988), S. 829-833 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: We have recently prepared some crosslinked derivatives of Escherichia coli β-galactosidase by treating the enzyme with bisimidoesters. In this article, we report the results obtained when the native and these crosslinked derivatives are entrapped in polyacrylamide gel lattice. It was found that use of combination of three protective agents, viz., bovine serum albumin, cysteine, and lactose, during immobilization gave an increased yield of 190% in the case of DMA crosslinked preparation. In the case of native enzyme, the Km, pH optimum, and temperature optimum were found to remain unchanged on immobilization. The DMA crosslinked preparation entrapped in polyacrylamide in the presence of BSA, lactose, and cysteine was found to be a significantly better catalyst and hydrolyzed 47% milk lactose as compared to 31% hydrolysis by entrapped native enzyme in 6 h.
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  • 55
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Notes: Among various microbial cells examined under screening conditions, Nocardia opaca showed the highest activity for production of phenylalanine from phenylpyruvate. Here NH4Cl as well as amino acids were used as an amino donor for phenylalanine production. The phenylalanine production rate increased with increasing hydrogen pressure. The specific activity of phenylalanine dehydrogenase was increased by culturing N. opaca cells in nutrient broth containing 0.3% phenylalanine. As a result, the phenylalanine production rate increased from 0.69 to 4.4 μmol/min g dry cells. Immobilized cells were activated in nutrient broth containing ZnCl2 before phenylalanine production. Phenylalanine dehydrogenase activity and cell number in the gel increased with increasing incubation time, and the maximum phenylalanine dehydrogenase activity was obtained at 36 h incubation. Then, phenylalanine was produced from phenylpyruvate, NH4Cl, and 100 atm H2 with the activated immobilized cells. The rate of phenylalanine production was 0.24 μmol/min cm3 gel. The conversion of phenylpyruvate to phenylalanine was 82%. Immobilized cells retained 76% of the initial phenylalanine production rate after 10 h reactions were repeated 11 times with two intervening reactivations.
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    Biotechnology and Bioengineering 32 (1988), S. 460-466 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Experimental data from the enzymatic hydrolysis of phenol-pretreated Swedish wheat straw have been used to evaluate the cost fractions of capital and utility, enzyme, and raw material. Two different raw material prices and varying enzyme prices have been used. The evaluation is based on an empirical model for the enzymatic hydrolysis and a computer program where utility and equipment, enzyme, and raw material prices can be varied. The optimal residence time for the enzymatic hydrolysis is in the range of 70-110 h. A fed-batch procedure with substrate concentrations higher than 10% oven-dried material (ODM) and enzyme concentrations in the range (6-10) · 106 FPU/ton ODM should be used.
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    Biotechnology and Bioengineering 32 (1988), S. 467-474 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: Antisera against charged (p-azobenzoate and p-azoben zenesulfonate) and uncharged (dinitrophenyl) haptenic groups were produced in rabbits, and the equilibrium characteristics of hapten-antibody were measured by use of immunoadsorbents. The antibody to the uncharged hapten formed a stable binding with the hapten to the changes in ionic strength and pH. On the other hand, the antibodies to the charged haptens showed affinities sensitive to the changes in pH and ionic strength. Therefore, the effect of the pKa of ionizable haptens on the pH dependence of the hapten-antibody binding was studied by comparing the interactions between a series of para-substituted benzoic acids and the anti-p-azobenzoate antibody. The pH dependence of the interactions was strongly affected by the pKa of ionizable groups in haptens. Furthermore, the equilibrium characteristics of anti-p-aminobenzoyl dipeptides were compared. The characteristics of interactions were affected by the features of amino acid residues.
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    Biotechnology and Bioengineering 32 (1988), S. 475-481 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A comparison of different immobilized enzyme reactors has been made for the limited modification of soya storage proteins and the products compared with those from action of the soluble enzyme. Clarified total water extracts of soya protein were subjected to the action of subtilisin in a soluble and immobilized form. The sodium dodecyl sulfate (SDS) electrophoresis patterns of soya proteins modified by enzyme in the two forms differed for unbuffered soya protein at the same pH of 8.0. However, identical patterns could be obtained by a downward adjustment of the pH of soya protein treated with immobilized enzyme. The same SDS electrophoresis pattern could be obtained for a packed column of immobilized enzyme and a well-mixed vessel by buffering. Operation of the column reactor at higher superficial linear velocities (above 1.47 cm/min), higher protein concentrations (8.8% w/v), and prolonged periods (24 h) led to a bed compression attributed to the protein coating of the support.
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  • 59
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    Biotechnology and Bioengineering 32 (1988), S. 482-490 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Empirical equations used so for for computing kla in mechanically agitated aerated vessels agree well with the measured dependence of Kla on the power input of the stirrer and the superficial velocity of the gas. However, they do not fit the dependence of Kla on the physical properties of the liquid batch. From the data published so far it can be deduced that the equations suitable for the description of the dependence of kmola on the liquid physical properties must include some independent variable, which characterizes the rate of coalescence of the bubbles. About 400 experimental values of Kla were compared with those calculated from different relations. This comparison shows that the rate of coalescence for the aqueous electrolyte solutions is well described on the basis of the Gibbs theory of adsorption of the solute in the surface layer of the solution.
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  • 60
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    Biotechnology and Bioengineering 32 (1988), S. 491-506 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The lack of quantitative descriptions of mammalian culture kinetics limits the ability to optimally design and control cell culture bioreactors. This limitation is ad dressed by developing mathematical equations relating the initial growth rate and the antibody productivity of the hybridoma cell line, CRL-1606, to its environmental state. This initial rate approach, in contrast with steady state analysis of chemostat cultures, approximates steady state behavior, since the rates were measured over brief time intervals and at low cell concentrations (〈 50,000 cells/mL). The advantage of this approach is that it is much faster than the chemostat approach.An equation for the growth rate was developed that superimpoed Monod equations in serum and glutamine with “noncompetitive” -type inhibition constants were inversely proportional to the lactate and ammonium concentrations. The Monod constant is critical for relating initial, low cell level culture states to other states.Lactate was found to be the only environmental parameter that significantly inhibited antifibronectin monoclonal antibody production by the CRL-1606 hybridimas. Volumetric productivity was strictly related to culture viability, which was observed to decline at growth rates below 0.02h-1. Lactate was also found to significantly inhibit ammonium production.
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  • 61
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Protocols for horizontal two-dimensional electrophoresis with immobilized pH gradients in the first dimension were modified for horizontal micro two-dimensional electrophoresis using PhastSystem. Different equilibration conditions of the first-dimensional immobilized pH gradient gel strip prior to second-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis were evaluated. Silver stained two-dimensional patterns were obtained within 3.5 h.
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  • 62
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    Electrophoresis 9 (1988), S. 47-53 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Rats were fed liquid food containing ethanol in concentrations ranging from 1-5 % for 13 weeks. Livers were removed for histopathology and the liver cytosolic protein fraction was prepared and used for two-dimensional gel electrophoresis (2D-PAGE). Polypeptides were visualized by silver staining. Scanning was made for estimation of the relative abundance of protein in each polypeptide spot in the gels and for comparison between rats. Visual inspection and scanning of gels with the stained polypeptide spots obtained after equilibrium isoelectric focusing and non-equilibrium pH gradient electrophoresis revealed that: (1) within the control rat and ethanol-treated rat livers the numbers of polypeptide spots detected using isoelectric focusing in the first dimension were approximately 500 and for non-equilibrium pH gradient electrophoresis 400; (2) in the control group the variation in the estimated amount of protein in each spot was remarkably small; (3) pronounced differences in the relative abundance of protein in several of the spots was observed in the ethanol-exposed rats as compared to controls. Dose-response relations and possible causes for the effects of ethanol are discussed.
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    Electrophoresis 9 (1988), S. 354-355 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A novel approach to the immunological stage of immunoblotting is described. The method employs the phenomenon of electroendosmosis to bring the antibodies sequentially into contact with the antigen. Using a three-stage peroxidase antiperoxidase system for human group specific protein (Gc) phenotyping, it has been shown that the method has comparable sensitivity, is more rapid and is more economical of antibodies than a similar multi-stage immerse-and-wash system.
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    Electrophoresis 9 (1988), S. 356-356 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Electrophoresis 9 (1988), S. 357-358 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 66
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    Electrophoresis 9 (1988), S. 485-488 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A new sample applicator for horizontal flat gels is described. The applicator is practically safe against contamination from adjacent samples and can be used for all types of electrophoretic separations including a concentration step for either the sample (i. e. disc electrophoresis) or the separated zones (i. e. isoelectric focusing). The applicator is a piece of flat glass with 26 or 51 parallel 2 mm wide grooves, drilled at distances of 9 or 4.5 mm. Samples, maximally 25 or 50. are applied to the areas between the grooves. By inverting the applicator, the samples are brought into close vicinity to the gel surface and the pendant droplets expand by capillary attraction into the slits between the glass and gel with resultant even distribution across the lanes of 2.5 or 7 mm width. The applicator can be used for separations with and without protection of the electrophoretic setup by paraffin oil and allows for fast multiple handling of samples by means of appropriate syringes and microtiter plates.
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  • 67
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Several procedures for casting 100 μm ultrathin immobilized pH gradients are described. When acrylate/glass molding cassettes (Pascali et al., Electrophoresis 1987, 8, 371-373) are used two main problems are encountered: (i) a tendency of polymerization solutions, at the beginning of delivery, to spread across the glass surfaces with troublesome effects on the gradient stratification, and (ii) the raising of steep menisci at both extremities of the pH intervals, originating from capillarity phenomena and resulting in nonuniform gradients with bowed electrophoretic patterns. The first shortcoming was acceptably solved by increasing the density of sucrose gradients, and pouring them into prewarmed molding cassettes. The detrimental effect of menisci could be overcome by using a ‘squeezing-sealing mold’ technique. A molding cassette was endowed with a continuous, squared spacer frame, the upper side being open by inserting a wedged clip. A slight excess of polymerization solution was first dispensed into the cassette and squeezed away on removal of the clip. By completely excluding air from the molding cassette, uniform and well reproducible ultrathin gels could be cast. A major advantage of ultrathin immobilized pH gradient gels is the drastically shorter focusing time.
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  • 68
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    Topics: Biology , Chemistry and Pharmacology
    Notes: A new modification of silver staining of proteins in sodium dodecyl sulfate polyacrylamide gels is adapted to automated staining in PhastSystem Development Unit. The use of a reduction step, after fixation, with thiosulfate in alcoholic sodium acetate buffer results in a considerable increase in sensitivity without the need for a recycling step. The detection limit is tenfold lower than in the silver staining procedure recommended so far for PhastSystem and corresponds to 0.05-0.1 ng protein per band. Total staining time with the new procedure is 75 min.
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  • 69
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Horizontal two-dimensional (2-D) electrophoresis with immobilized pH gradients (IPG) in the first dimension for buffer soluble proteins and for complex proteins solubilized in the presence of Nonidet P-40 (Görg et al., Electrophoresis 1987, 8, 45-51), has been extended to analyze basic proteins of yeast cells focused under non-equilibrium and equilibrium conditions. Transient state isoelectric focusing (IEF) in IPG gels revealed sample smearing and background staining, displaying horizontal streaks in the resultant 2-D patterns. Inclusion of 0.5 % carrier ampholytes (CA) to the IPG gel (IPG-CA), resulted in the formation of many sharp protein bands after transient state IEF with resultant distinct spots in the 2-D patterns; however, resolution was poor and the gel contained heavy background staining. With prolonged focusing time, background staining disappeared and there was less difference in the final steady state IEF patterns obtained with IPG and IPG-CA. Reduction of the Immobiline concentration to one third the manufacturer's recommended amount did not improve IEF resolution with respect to streaking and background staining under either transient state or equilibrium conditions. In general, spot intensities were less on 2-D gels using diluted IPG gels than with “standard” IPG gels. Optimization of 2-D electrophoresis with IPGs in the first dimension was strongly related to IEF conditions. The use of IPG gels focused to equilibrium should not only improve inter-gel reproducibility and resolution but also the quality of the final 2-D patterns with respect to background staining and horizontal streaking.
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    Electrophoresis 9 (1988), S. 839-841 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The practical usefulness of multicolumn isotachophoresis was demonstrated by the determination of oxalate levels in human serum. 10 mmol/L HC1 and 100 mmol/L Na2HPO4 served as the leading and terminating electrolyte, respectively. For the stabilization of the isotachophoretic zones in large cross section channels a suspension of Bio-Gel P-300 in the leading electrolyte was used. For the analysis ca. 1 mL of fresh serum was required and 1.2 mL of 1:1 mixture of serum and the suspension of Bio-Gel P-300 in deionized water was applied as sample. The time of analysis ranged between 45-49 min. The detection limit of analysis was determined to be 50 nmol/L. Reproducibility of analysis of 1 μmol/L oxalate in water standard was found to be 2.6% (n = 6).
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    Electrophoresis 9 (1988), S. 846-846 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Electrophoresis 9 (1988), S. 846-846 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Electrophoresis 9 (1988), S. 291-291 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Electrophoresis 9 (1988), S. 292-292 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Electrophoresis 9 (1988), S. 303-306 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The 45Ca filtration method which allows calcium-binding proteins to be detected after their adsorption to nitrocellulose filters was used to test conditions for improved Western blotting of calmodulin. Our results indicate that dodecyl sulphate is not efficiently removed from calmodulin during electroelution in standard transfer buffers. This detergent disrupts binding of calmodulin to nitrocellulose. Electrotransfer in potassium buffer which precipitates dodecyl sulphate leads to a 6-fold increase in calmoculin retention by nitrocellulose. With this and other modifications, the detection limit of the 45Ca overlay method of Maruyama et al. (J. Biochem. 1984, 95, 511-519) for calmodulin is increased to 0.2 μg per lane.
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    Electrophoresis 9 (1988) 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Electrophoresis 9 (1988), S. 380-383 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Genetic polymorphism of orosomucoid (ORM) has been demonstrated in several populations and comprises two structural gene loci, ORM 1 and ORM2. In Caucasians three common ORM 1 alleles have been shown, while the ORM 2 locus is almost monomorphic. ORM 1 phenotyping by isoelectric focusing in agarose or polyacrylamide gel combined with either print immunofixation or enzyme-linked immunoblotting is described, and population and family data from Denmark and Southern Germany are given. It is proposed to use a different alpha-numerical nomenclature for the phenotypes of the ORM 1 and ORM 2 systems.
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    Electrophoresis 9 (1988), S. 569-575 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Apolipoproteins C are involved in many ways in the metabolism of plasma lipoproteins. Apolipoproteins C from the delipidated VLDL of 35 controls and 165 normo-and hyperlipoproteinemic patients were analyzed by isoelectric focusing on an immobilized pH gradient, pH 4.0-5.0, with 7 M urea, which raised the apparent pH range to 4.8-5.7. This method is an improvement over conventional isoelectric focusing with carrier ampholytes with regard to both resolution and reproducibility. Due to the high resolution (0.1 pH units per cm) additional apolipoprotein C-III bands: C-III0 A1, C-III0 A2, C-III1 C and C-III2 C (the designations A, anodic, and C, cathodic, refer to direction of migration on IEF in relation to the main band) are described for the first time. The possible artifactual nature of these protein bands could be excluded. Cleavage with neuraminidase and peptidases, immunological detection and/or two-dimensional electrophoresis were used to obtain more information. The additional bands seem, in part, to be hydrolysis products of carboxypeptidase A (C-III1 C, C-III2 C). The appearance of C-III1 C and C-III2 C was dependent upon the serum triglyceride concentration. The percent distribution of C apolipoproteins in very low density lipoproteins (VLDL) from control serum agreed with previously published data. Apolipoproteins C can also be focused in immobilized pH gradients from VLDL and serum without delipidation.
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  • 79
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    Topics: Biology , Chemistry and Pharmacology
    Notes: The heterogeneity of human transferrin results from (i) differences in iron content, (ii) genetic polymorphism and (iii) differences in the carbohydrate moiety. This article primarily deals with the last phenomenon, the microheterogeneity of human transferrin. Owing to the comparatively simple carbohydrate structure of human transferrin and the high resolving power of isoelectric focusing in immobilized pH gradients, microheterogeneous forms of transferrin can be separated. Differences between samples can be quantitated by crossed immunoelectrophoresis. Examples of the differences between the microheterogeneity patterns of transferrin in several biological fluids and the changes that can be observed in diseases such as rheumatoid arthritis, idiopathic hemochromatosis and Kahler's disease are presented. Special attention has been focused on changes occurring during pregnancy.
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  • 80
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    Topics: Biology , Chemistry and Pharmacology
    Notes: The genetic variants of bovine β-lactoglobulin (β-lg) from the “Murnau-Werdenfelser” breed were analyzed in three different isoelectric focusing (IEF) systems. While carrier ampholyte IEF with a pH gradient of 0.2 pH/cm did not resolve the new variant W from the B variant and IEF in immobilized pH gradients (IPG) with 0.1 pH/cm only partially resolved it, adequate separation was achieved with IPG-IEF in a pH 5.25-pH 5.7 gradient, in presence of 0.8 % w/v carrier ampholytes, both over a 10 and 17 cm separation distance. Apparent isoelectric points (pI's) and genetic frequencies (f) were as follows: β-lg A, pI = 5.370,f = 0.364; β-lg B, pI = 5.485,f = 0.480; β-lg W, pI = 5.492,f = 0.076; and β-lg D, pI = 5.610,f = 0.080. The small difference of Δ pI = 0.007 between β-lg B and β-lg W respectively, seems to originate from a “silent” substitution of neutral amino acid residues as compared to the larger Δ pI's of the other genetic variants of β-lg, which result from substitution of charged amino acids.
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    Electrophoresis 9 (1988) 
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    Electrophoresis 9 (1988), S. 654-661 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A sensitive method is described for the detection of tissue peptides and proteins. They are separated by tissue isoelectric focusing using thin large-pore polyacrylamide gels, containing detergent and dimethylformamide, and are fixed with either glutaraldehyde or formaldehyde in gelatin-coated nitrocellulose membranes using press-blotting. The fixed peptide and protein antigens are visualized by immunoperoxidase staining. The spectrum of fixed tissue constituents may also be used to test antiserum reactivity and specificity in immunocytochemical staining procedures. Isoelectric focusing of 2 μL homogenates of the neurointermediate lobe of the pituitary allowed the immunodetection of peptides and proteins of various sizes and the determination of isoelectric points. However, direct application onto gels of small pieces of frozen tissue sections, sliced in a cryostat, appeared to be more efficient. By direct tissue isoelectric focusing of brain tissue, peptides were effectively eluted and separated from sections up to 100 μm thickness. This allowed the detection of small peptides with a detection limit of approximately 10 pg/section.
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    Notes: High resolution two-dimensional electrophoresis indicates that serum proteins previously only detected in patients with acute myocardial infarction (AMI) (Gomo et al., Electrophoresis 1983, 4, 298-302) are also present in the serum protein patterns of other patients (AMI negative or demonstrating acute phase response) and are faintly detected even in controls. Thus, these proteins are not specific to AMI and are probably acute phase reactants. However, they do demonstrate a characteristic time course response in sequential samples from AMI patients.
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    Electrophoresis 9 (1988), S. 679-679 
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    Electrophoresis 9 (1988), S. 742-750 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Using high-resolution two-dimensional polyacrylamide gel electrophoresis we studied the polymorphism of protein amounts in some genotypes of maize and pea. This type of variability seems to be rather common and insensitive to environmental conditions, as attested by the comparison of the patterns of two maize lines harvested in two different years. A large-scale experiment involving 5 lines, 7 of their hybrids, and 6 organs (or physiological stages) of maize allowed us to examine numerous polypeptides regrading their genetic variability, their amount differences between organs and the inheritance of their abundance. Genetic and organ variations are not independent: polypeptides whose amount varies from one organ to another are, for the most part, genetically variable (59 %), while the stable polypeptides are not often genetically variable (18 %). We found a striking organ specificity for (i) the extent of quantitative variability (from 2.3-15.4 % of the polypetides), (ii) the occurrence and the type of variation for a given polypeptide (an intensity difference seen in an organ can disappear or even be reversed in another one), (iii) the kind of inheritance (additive/non-additive): combining the 6 organs and the 7 hybrids we found 101 cases of non-additivity (4 % of the total) which conecern as many as 72 different spots, that is to say that in most cases a polypeptide displaying nonadditivity in an organ seems to display additivity in the other ones. Moreover, for most of the polypeptides with nonadditive inheritance the hybrid spot presents an intensity similar to that of the most intense parental spot. Thus for protein experssion a hybrid cannot be predicted from its parents. The organ-specificity was also evidenced by analyzing pleiotropic effect of gene substitutions in various organs of garden pea. Comparisons of isogenic lines differing for the r-locus revealed that according to the organ, the gene substitution caused appearances, disappearances or quantitative changes for 0 % to more than 10 % of the proteins revealed. All these features are discussed in connection with the possible role of variation of protein amounts in phenotypic variability.
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    Electrophoresis 9 (1988), S. 796-796 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 87
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    Notes: An improved method for the estimation of molecular weights of native proteins by polyacrylamide gel electrophoresis, in 9 cm × 9 cm × 0.05 mm 4-20% T fabric reinforced gradient gels, is described. Plotting the logarithm of the relative mobilities of proteins versus gel concentrations produces lines whose slopes are related to molecular weights.
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  • 88
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    Notes: The use of a bivalent counter anion for cationic isotachophoresis was studied both theoretically and experimentally. A mathematical model was proposed and solved to give the effective mobility of the hydrogen ion constituent, uH.H, in the respective electrolyte systems. The theoretical values agreed well with the experimental data in a series of electrolyte systems. The dependence of uH.H on such parameters as pH of the leading electrolyte, ionic mobilities of leading and counter ions, and dissociation constants was calculated and discussed. A bivalent counter anion will prove useful for the separation of low mobility cations and weak bases, as shown for the separation of substituted anilines in a succinate electrolyte system.
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    Electrophoresis 9 (1988), S. 816-819 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Human lactate dehydrogenase (LDH) was investigated by ultrathin-layer polyacrylamide gel isoelectric focusing (IEF). In serum, approximately 15 bands and in lyzates of erythrocytes approximately 20 bands were detected. Known LDH isoenzymes (identified by markers) appeared in the zymograms as follows: LDH-1 as a single or double band, LDH-2 as a single band in serum and in the marker, and as a double band in hemolyzate, LDH-3 as a double band, and LDH-4 and LDH-5 each as a single band. LDH-1 was partly inactivated, probably due to deamidation in the acidic range of the pH gradient. Potential LDH tumor markers were detected in different tumor cytosols.
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    Electrophoresis 9 (1988), S. 841-844 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Plant glycoproteins generally cross-react because of the presence of identical or related complex glycans which are highly immunogenic. The use of mild periodate oxidation of glycans after glycoprotein transfer from sodium dodecyl sulfate-poly-acrylamide gel electrophoresis gels to nitrocellulose membranes prior to immunodetection is a way of identifying the carbohydrate antigenic determinants of a glycoprotein as the basis for antigenic cross-reaction. Periodate oxidation can distinguish between antibodies directed against carbohydrate and against peptide antigenic determinants, the latter being unaffected by oxidation. Immunoblotting performed after periodate treatment allows the detection of common protein epitopes.
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  • 91
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 9 (1988), S. 751-763 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Holoprotein molecular weights and polypeptide composition can be determined for complex mixtures of oligomeric proteins using two-dimensional electrophoretic techniques. The variety of two-dimensional analyses presented here is a reflection of the general usefulness of each method for the identification and characterization of the different classes of seed storage proteins in alfalfa. These techniques can be applied to studies of storage proteins in other seeds as well as non-seed storage proteins. The major seed storage proteins in alfalfa are medicagin (a legumin-like globulin), alfin (a vicilin-like globulin) and a family of lower molecular weight albumins (LMW1-3). These comprise 30 %, 10 %, and 20 %, respectively, of the total extractable protein from cotyledons of mature seeds. Alfin is a heterogeneous oligomeric protein (Mr ∼ 150 000) composed of polypeptides ranging in size from Mr 14 000 to 50 000 (α1-α6; 50 000, 38 000, 32 000, 20 000, 16 000 and 14 000, respectively). Medicagin is also a high molecular weight oligomeric protein, but requires high concentrations of salt for solubilisation. It is comprised of a family of individually distinct subunits, each composed of an acidic polypeptide (A1-A9; Mr 49 000 to 39 000) linked via disulphide bond(s) to a basic polypeptide (B1, B2, B3; Mr 24 000, 23 000 and 20 000, respectively). This pairing is highly specific and two families are recognizable on the basis of the B polypeptide (B3 or B1 /B2). Subunits (Mr ∼ 50 000-65 000) are assembled as trimers (8S) or larger oligomers (12S-15S) in mature seeds. The lower molecular weight albumins (LMW1-3) are acidic (pI 〈 6), and consist of sets of disulphide-bonded polypeptides (Mr 15 000 and 11 000).
    Additional Material: 12 Ill.
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  • 92
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 9 (1988) 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 93
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Sodium dodecyl sulphate-polyacrylamide gel electrophoresis has been used to determine the difference between thylakoid polypeptide patterns of light and dark grown strains of the cyanobacterium Chlorogloea fritschii. There were only 2 prominent bands present in the dark grown strains, polypeptide Mr 50 000 and polypeptide Mr 90 000, also five fainter bands in the Mr range 45 000-66 200 corresponding to photosystem one, compared with the 32 bands present in the light grown strains. There was no obvious indication of the Mr 33 000 3-(3,4-chlorophenyl)-1,1-dimethyl-urea binding protein. In addition the progressive daily development of the various photosystem components in the light and their relationship in photosynthesis was determined. It was observed that the increase of the relative concentrations of the photosystem two and phycocyanin components indicated their developments are mutually synchronized. The effect of light to dark and dark to light transfer on established strains was investigated. Appreciable loss of photosystem two components and the presence of an additional band Mr 22 500 of unknown function in the light to dark transfer, and little reactivation of the photosynthetic capabilities in the dark to light transfer was observed.
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  • 94
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The development of equipment for fast automated staining is described. It is possible to handle staining procedures with up to 20 steps and nine different solutions. To increase the reaction rate in the reaction chamber, the gels are rotated and high temperatures are used. The temperature in the reaction chamber is controlled between room temperature and 50 °C. Increased temperature, above 20 °C, generally results in faster staining and destaining. However, some reactions proceed better at a low temperature, including fixation of proteins with TCA, and the development step in silver staining, where increased temperatures cause a high background stain. Silver in silver staining, where increased temperatures cause a high background stain. Silver staining using acidic silver nitrate solution is preferred, due to easy preparation and good storage stability of the reagents. This method also causes little precipitation of silver on the walls of the reaction chamber. Silver staining is accomplished within one hour. Staining with PhastGel Blue is accomplished within 30 min.
    Additional Material: 11 Ill.
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  • 95
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 9 (1988), S. 60-61 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Staining of proteins in PhastGel gradient media with Coomassie Blue R 350 was considerably improved using a lower concentration of methanol (10% v/v) and 2% ammonium sulfate in the staining solution and 10% acetic acid for destaining. The detection limit in sodium dodecyl sulfate-polyacrylamide gels was lowered by a factor of 10 to about 2 ng per protein band. The Coomassie staining method was adapted to the newly developed silver staining procedure so that both can be used in parallel in PhastSystem.
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  • 96
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The effect of salt and buffer ions in the sample or in an immobilized pH gradient (IPG) on sample entry into the gel and on the final focused pattern are presented. During the initial phase of electrofocusing, ions present in the gel, either as counter ions to the immobilized charge groups of the IPG gel or added to the gel matrix during the rehydration process, are transported toward the electrodes. For ions present at a concentration exceeding ∼ 1 mM the transport can be followed by the refractile line marking the trailing edge of an ion-containing zone. Gradual sample entry may be achieved by applying the sample at a site (near the anode or cathode) opposite to that from which the sharpest refractile line, marking the ion present in the highest concentration, approaches the sample. Additionally, lateral band spreading of the sample is avoided. Thus, sample applied at the cathode for IPG gels rehydrated with 1-2 mM Tris base, or at the anode for gels rehydrated with 1-2 mM acetic acid or sodium acetate, enters the gel matrix gradually without lateral band spreading. In contrast, sample applied at the anode, for Tris-containing gels, or at the cathode, for acetate-containing gels, enters rapidly in a sharp zone when the refractile line reaches the sample zone. This results in a high local protein concentration in the zone immediately behind the boundary with lateral band spreading. Salt concentrations in excess of ∼ 0.15 M in the sample result in electroosmotic removal of water from the sample before the proteins can enter the gel, with resultant precipitation of the sample at the application site. Local pH extremes can be created at the application point, which may produce delayed sample entry or protein precipitation.
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  • 97
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Improved methods for high resolution composite gel electrophoresis under dissociating conditions and electrophoretic transfer of immunoglobulins (Mr 150 000-1 800 000) to nitrocellulose have been developed. Using these techniques and highly specific antisera to detect either light or heavy chains on nitrocellulose transfers, the immunoglobulin and clonal composition of washed cryoglobulins (3 μg each) could be determined within two days. The results were confirmed by isoelectric focusing and immunoblotting of selected samples. In addition, the method was used for detection of andiotypic determinant and rheumatoid factor activity within components of the cryoglobulins.
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  • 98
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The interpretation of two-dimensional gel electrophoresis (2-DGE) profiles can be facilitated by artificial intelligence and machine learning programs. We have incorporated into our 2-DGE computer analysis system (termed MELANIE-Medical Electrophoresis Analysis Interactive Expert system) a program which automatically classifies 2-DGE patterns using heuristic clustering analysis. This program is a step toward machine learning. In this publication, we describe the classification method and the preliminary results obtained with liver biopsy electrophoretograms. Heuristic clustering is also compared to other classification techniques.
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  • 99
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 9 (1988) 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 100
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Hydrophobic affinity electrophoresis under high hydrostatic pressure has been developed to study the interaction between fatty acid-free bovine serum albumin and a long-chain aliphatic ligand physically immobilized within the gel matrix. From apparent association constants at various pressures and temperatures, apparent thermodynamic parameters including the volume change in binding were calculated. The results are as expected for hydrophobic interactions between the long-chain alkyl ligand and a high-affinity long-chain fatty acid binding site. The feasibility of high-pressure affinity electrophoresis is demonstrated. This new high-pressure technique provides a direct means for studying quantitatively the effects of pressure upon protein-ligand interactions. It could become a suitable tool for the investigation of protein binding sites' topography.
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