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  3. Biotechnology and Bioengineering
  4. 1993
Showing papers in "Biotechnology and Bioengineering in 1993"
Journal Article•10.1002/BIT.260410808•
Biosorption of cadmium by biomass of marine algae

[...]

Z. R. Holan1, Bohumil Volesky1, Imam Prasetyo1•
McGill University1
05 Apr 1993-Biotechnology and Bioengineering
TL;DR: Biomass of nonliving, dried brown marine algae Sargassum natans, Fucus vesiculosus, and Ascophyllum nodosum demonstrated high equilibrium uptake of cadmium from aqueous solutions and there was no damage to the biosorbent which retained its macroscopic appearance and performance in repeated metal uptake/elution cycles.
Abstract: Biomass of nonliving, dried brown marine algae Sargassum natans, Fucus vesiculosus, and Ascophyllum nodosum demonstrated high equilibrium uptake of cadmium from aqueous solutions. The metal uptake by these materials was quantitatively evaluated using sorption isotherms. Biomass of A. nodosum accumulated the highest amount of cadmium exceeding 100 mg Cd[sup 2+]/g (at the residual concentration of 100 mg Cd/L and pH 3.5), outperforming a commercial ion exchange resin DUOLITE GT-73. A new biosorbent material based on A. nodosum biomass was obtained by reinforcing the algal biomass by formaldehyde cross-linking. The prepared sorbent possessed good mechanical properties, chemical stability of the cell wall polysaccharides and low swelling volume. Desorption of deposited cadmium with 0.1-0.5 M HCl resulted in no changes of the biosorbent metal uptake capacity through five subsequent adsorption/desorption cycles. There was no damage to the biosorbent which retained its macroscopic appearance and performance in repeated metal uptake/elution cycles.

546 citations

Journal Article•10.1002/BIT.260420203•
A mathematical model for dynamic simulation of anaerobic digestion of complex substrates: focusing on ammonia inhibition.

[...]

Irini Angelidaki1, L. Ellegaard1, Birgitte Kiær Ahring1•
Technical University of Denmark1
20 Jun 1993-Biotechnology and Bioengineering
TL;DR: A mathematical model for anaerobic degradation of complex organic material, such as manure, has been developed and results compare favorably with experimental data.
Abstract: A mathematical model for anaerobic degradation of complex organic material, such as manure, has been developed. The model includes an enzymatic hydrolytic step and four bacterial steps and involves 12 chemical compounds. The model focuses on ammonia inhibition and includes a detailed description of pH and temperature characteristics in order to accurately simulate free ammonia concentration. Free ammonia and acetate constitute the primary modulating factors in the model. The model has been applied for the simulation of digestion of cattle manure in continuously stirred tank reactors (CSTRs), and results compare favorably with experimental data.

457 citations

Journal Article•10.1002/BIT.260420811•
Activity studies of eight purified cellulases: Specificity, synergism, and binding domain effects.

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Diana C. Irwin1, Michael Spezio1, Larry P. Walker1, David Wilson1•
Cornell University1
01 Oct 1993-Biotechnology and Bioengineering
TL;DR: The activities of six purified Thermomonospora fusca cellulases and Trichoderma reesei CBHI and CBHII were determined on filter paper, swollen cellulose, and CMC.
Abstract: The activities of six purified Thermomonospora fusca cellulases and Trichoderma reesei CBHI and CBHII were determined on filter paper, swollen cellulose, and CMC. A simple method to measure the soluble and insoluble reducing sugar products from the hydrolysis of filter paper was found to effectively distinguish between exocellulases and endocellulases. Endocellulases produced 34% to 50% insoluble reducing sugar and exocellulases produced less than 8% insoluble reducing sugar. The ability of a wide variety of mixtures of these cellulases to digest 5.2% of a filter paper disc in 16 h was measured quantitatively. The specific activities of the mixtures varied from 0.41 to 16.31 micromol cellobiose per minute per micromole enzyme. The degree of synergism ranged from 0.4 to 7.8. T. reesei CBHII and T. fusca E3 were found to be functionally equivalent in mixtures. The catalytic domains (cd) of T. fusca endocellulases E2 and E5 were purified and found to retain 93% and 100% of their CMC activity, respectively, but neither cd protein could digest filter paper to 5.2%. When E2cd and E5cd were substituted in synergistic mixtures for the native proteins, the mixtures containing E2cd retained 60%, and those containing E5cd retained 94% of the original activity. Addition of a beta-glucosidase was found to double the activity of the best synergistic mixture. Addition of CBHI to T. fusca crude cellulase increased its activity on filter paper 1.7-fold.

369 citations

Journal Article•10.1002/BIT.260410308•
Formation of microparticulate protein powder using a supercritical fluid antisolvent

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Sang‐Do ‐D Yeo1, Gio-Bin Lim1, Pablo G. Debendetti1, Howard Bernstein•
Princeton University1
05 Feb 1993-Biotechnology and Bioengineering
TL;DR: In vivo tests on rats revealed no differences between the biological activity of processed and unprocessed insulin, and GAS processing of organic solution appears to be a reliable and effective method for the production of dry, biologically active microparticulate powders of peptides and proteins.
Abstract: Gas antisolvent (GAS) expansion of dimethylsulfoxide (DMSO) and N,N-dimethylformamide (DMFA) solutions with supercritical carbon dioxide was used to produce biologically active powders of insulin. Powders with 90% of the particles smaller than 4 μm and 10% smaller than 1 μm were obtained under all conditions tested when the process was operated continuously, with small liquid droplets sprayed into a flowing supercritical continuum. Slow pressurization of the stagnant protein solution resulted in larger particles. In vivo tests on rats revealed no differences between the biological activity of processed and unprocessed insulin, GAS processing of organic solution appears to be a reliable and effective method for the production of dry, biologically active microparticulate powders of peptides and proteins. © 1993 John Wiley & Sons, Inc.

344 citations

Journal Article•10.1002/BIT.260410809•
Cadmium biosorption by Saccharomyces cerevisiae

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Bohumil Volesky1, H. May1, Z. R. Holan1•
McGill University1
05 Apr 1993-Biotechnology and Bioengineering
TL;DR: Nearly linear sorption isotherms featured by higher sorbing resting cells together with metal deposits localized exclusively in vacuoles indicate the possibility of a different metal-sequestering mechanism when compared to dry nonliving yeasts which did not usually accumulate more than 20 mg Cd/g.
Abstract: Cadmium uptake by nonliving and resting cells of Saccharomyces cerevisiae obtained from aerobic or anaerobic cultures from pure cadmium-bearing solutions was examined. The highest cadmium uptake exceeding 70 mg Cd/g was observed with aerobic baker's yeast biomass from the exponential growth phase. Nearly linear sorption isotherms featured by higher sorbing resting cells together with metal deposits localized exclusively in vacuoles indicate the possibility of a different metal-sequestering mechanism when compared to dry nonliving yeasts which did not usually accumulate more than 20 mg Cd/g. The uptake of cadmium was relatively fast, 75% of the sorption completed in less than 5 min.

324 citations

Journal Article•10.1002/BIT.260420415•
A thermodynamically based correlation for maintenance gibbs energy requirements in aerobic and anaerobic chemotrophic growth.

[...]

L. Tijhuis1, M.C.M. van Loosdrecht1, J. J. Heijnen1•
Delft University of Technology1
05 Aug 1993-Biotechnology and Bioengineering
TL;DR: The generalized concept for maintenance Gibbs energy is used to establish a correlation which allows the estimation of the biomass yield on electron donor as a function of C‐source, electron donor, electron acceptor, N source, growth rate, and temperature.
Abstract: A thermodynamic framework has been provided for the description of maintenance requirements of microorganisms. The central parameter is the biomass specific Gibbs energy consumption for maintenance, m(E) (kJ/C-mol biomass . h). A large set of data has been used including (i) a large range of different organisms (bacteria, yeasts, plant cells), (ii) mixed cultures, (iii) heterotrophic and autotrophic growth, (iv) growth under aerobic and anaerobic conditions, and (v) a large temperature range (5-75 degrees C). It appears that only the temperature has a major influence, with an energy of activation of 69 kJ/mol. Different electron donors or electron acceptors only show a very minor influence on m(E). On the basis of the data set, temperature correlations of m(E) have been derived for aerobic and anaerobic growth. The generalized concept for maintenance Gibbs energy is used to establish a correlation which allows the estimation of the biomass yield on electron donor as a function of C-source, electron donor, electron acceptor, N source, growth rate, and temperature. The advantage of using the m(E) parameter over other maintenance-related parameters (like mu(e), m(O2), m(D), gamma(D)m(D)) is discussed.

293 citations

Journal Article•10.1002/BIT.260420509•
Effect of surfactants on cellulose hydrolysis

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Steve S. Helle1, Sheldon J.B. Duff1, David G. Cooper2•
University of British Columbia1, McGill University2
20 Aug 1993-Biotechnology and Bioengineering
TL;DR: Sophorolipid had the greatest effect on cellulose hydrolysis when it was present from the beginning of the experiment and when the enzyme/cellulose ratio was low.
Abstract: The effect of surfactants on the heterogeneous enzymatic hydrolysis of Sigmacell 100 cellulose and of steam-exploded wood was studied. Certain biosurfactants (sophorolipid, rhamnolipid, bacitracin) and Tween 80 increased the rate of hydrolysis of Sigmacell 100, as measured by the amount of reducing sugar produced, by as much as seven times. The hydrolysis of steam-exploded wood was increased by 67% in the presence of sophorolipid. At the same time, sophorolipid was found to decrease the amount of enzyme adsorbed onto the cellulose at equilibrium. Sophorolipid had the greatest effect on cellulose hydrolysis when it was present from the beginning of the experiment and when the enzyme/cellulose ratio was low.

288 citations

Journal Article•10.1002/BIT.260420615•
Removal of lead from aqueous solutions by Penicillium biomass.

[...]

Hui Niu, Xue Shu Xu, Jian Hua Wang, Bohumil Volesky1•
McGill University1
05 Sep 1993-Biotechnology and Bioengineering
TL;DR: The removal of lead ions from aqueous solutions by adsorption on nonliving Penicillium chrysogenum biomass was studied in this paper, where the saturated sorption uptake of Pb(+2) was 116 mg/g dry biomass, higher than activated charcoal and some other microorganisms.
Abstract: The removal of lead ions from aqueous solutions by adsorption on nonliving Penicillium chrysogenum biomass was studied. Biosorption of the Pb(+2) ion was strongly affected by pH. Within a pH range of 4 to 5, the saturated sorption uptake of Pb(+2) was 116 mg/g dry biomass, higher than that of activated charcoal and some other microorganisms. At pH 4.5, P. chrysogenum biomass exhibited selectivity for Pb(+2) over other metal ions such as Cd(+2), Cu(+2), Zn(+2), and As(+3) Sorption preference for metals decreased in the following order: Pb > Cd > Cu > Zn > As. The sorption uptake of Pb(+2) remained unchanged in the presence of Cu(+2) and As(+3), it decreased in the presence of Zn(+2), and increased in the presence of Cd(+2).

277 citations

Journal Article•10.1002/BIT.260410503•
Biofiltration of methanol vapor.

[...]

Zarook Shareefdeen1, Basil C. Baltzis, Young‐Sook ‐S Oh2, Richard Bartha2•
New Jersey Institute of Technology1, Rutgers University2
05 Mar 1993-Biotechnology and Bioengineering
TL;DR: Both experimental data and model predictions suggest that the meethanol biofiltration process was limited by oxygen diffusion and methanol degradation kinetics.
Abstract: Biofiltration of solvent and fuel vapors may offer a cost-effective way to comply with increasingly strict air emission standards. An important step in the development of this technology is to derive and validate mathematical models of the biofiltration process for predictive and scaleup calculations. For the study of methanol vapor biofiltration, an 8-membered bacterial consortium was obtained from methanol-exposed soil. The bacteria were immobilized on solid support and packed into a 5-cm-diameter, 60-cm-high column provided with appropriate flowmeters and sampling ports. The solid support was prepared by mixing two volumes of peat with three volumes of perlite particles (i.e., peat-perlite volume ratio 2:3). Two series of experiments were performed. In the first, the inlet methanol concentration was kept constant while the superficial air velocity was varied from run to run. In the second series, the air flow rate (velocity) was kept constant while the inlet methanol concentration was varied. The unit proved effective in removing methanol at rates up to 112.8 g h(-1) m(-3) packing. A mathematical model has been derived and validated. The model described and predicted experimental results closely. Both experimental data and model predictions suggest that the methanol biofiltration process was limited by oxygen diffusion and methanol degradation kinetics.

275 citations

Journal Article•10.1002/BIT.260420610•
Toxicity of homologous series of organic solvents for the gram-positive bacteria Arthrobacter and Nocardia Sp. and the gram-negative bacteria Acinetobacter and Pseudomonas Sp.

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M.H. Vermuë, J. Sikkema, Annette Verheul, Rudolf Bakker, Johannes Tramper 
05 Sep 1993-Biotechnology and Bioengineering
TL;DR: The toxicity of homologous series of organic solvents has been investigated for gram‐positive bacteria, Arthrobacter sp.
Abstract: The toxicity of homologous series of organic solvents has been investigated for the gram-positive bacteria, Arthrobacter sp. and Nocardia sp., and the gram-negative bacteria, Acinetobacter sp. and Pseudomonas sp. The hydrophobicity of the solvent, expressed by its logP(octanol), proves to be a good measure for the toxicity of solvents in a two-phase system. The transition from toxic to nontoxic solvents occurs between logP(octanol) 3 and 5 and depends on the homologous series. No correlation has been found between the hydrophobicity of the substituent on the alkyl backbone of the solvent and the location of the transition point in toxicity. The logP(octanol), above which all solvents are nontoxic, is used to express the solvent tolerance of the bacteria. In general, the solvent tolerance of gram-negative bacteria is found to be slightly higher than that of gram-positive bacteria, but this does not hold for all homologous series of organic solvents investigated.Because the toxicity effects of organic solvents in a two-phase system can be ascribed to molecular as well as phase toxicity effects, molecular toxicity effects were investigated separately in a one-phase system with subsaturating amounts of organic solvent. The solvent concentration in the aqueous phase, at which 50% of the metabolic activity of the bacteria is lost, is used to express solvent toxicity. This concentration is found to be similar for the gram-positive Arthrobacter and the gram-negative Acinetobacter. Assuming the critical membrane concentration theory (G. J. Osborne et al. Enzyme Microb. Technol. 1990, 12: 281-291) to be valid, it can be concluded that differences in solvent tolerance between these two bacteria, cannot be ascribed to differences in response to molecular toxicity. Prediction of the toxicity of any solvent, using the critical membrane theory, appears to be possible in the case of alkanols or alkyl acetates. However, prediction of the toxicity of ethers appears to be impossible.

215 citations

Journal Article•10.1002/BIT.260410122•
Yield of poly-D(-)-3-hydroxybutyrate from various carbon sources: a theoretical study.

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Tsuneo Yamane1•
Nagoya University1
05 Jan 1993-Biotechnology and Bioengineering
TL;DR: The theoretical yield of poly‐D(‐)‐3‐hydroxybutyrate (PHB) has been estimated from the biochemical pathway leading to PHB when a carbohydrate (glucose), a C1 compound, a C2 compound, or a C4 compound is used as a carbon source.
Abstract: The theoretical yield of poly-D(-)-3-hydroxybutyrate (PHB) has been estimated from the biochemical pathway leading to PHB when a carbohydrate (glucose), a C(1) compound (methanol), a C(2) compound (acetic acid), or a C(4) compound (butyric acid) is used as a carbon source. In estimating the yield, recycling (or regeneration) of NADP(+)/ (NADPH + H(+)) and NAD(+) /(NADH + H(+)) have been taken into account. A special emphasis is made on te regeneration of NADPH, which is the coenzyme of acetoacetyl-CoA reductase, one of three key enzymes involved in the biosynthesis of PHB. As a NADPH-regenerating enzyme, glucose-6-phosphate dehydrogenase or isocitrate dehydrogenase is conceived. An equation which predicts the overall yield of PHB when non-PHB residual biomass is actually formed has been derived as a function of both the theoretical yield and PHB content of the dry cell mass. The ratio of the overall yield to the theoretical yield is roughly proportional to the PHB content.
Journal Article•10.1002/BIT.260411107•
The Kinetics of Cometabolism

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Craig S. Criddle1•
Michigan State University1
01 May 1993-Biotechnology and Bioengineering
TL;DR: Three previously proposed models describing the kinetics of cometabolism by resting cells are compared, and the interrelationships and underlying assumptions for these models are explored.
Abstract: Experimental observations indicate that the rates of cometabolic transformation are linked to the consumption of growth substrate during growth and to the consumption of cell mass and/or energy substrate in the absence of growth substrate. Three previously proposed models (models 1 through 3) describing the kinetics of cometabolism by resting cells are compared, and the interrelationships and underlying assumptions for these models are explored. Models 1 to 3 are shown to converge at high concentrations of the nongrowth substrate. An expression describing nongrowth substrate transformation in the presence of growth substrate is proposed, and this expression is integrated with an expression for cell growth to give a single unstructured model (model 4) that encompasses models 1 to 3 and describes cometabolism by both resting and growing cells. Model 4 couples transformation of nongrowth substrate to consumption of growth substrate and biomass, and predicts that cometabolism will result, and decreased specific growth rates for a cometabolizing population. Competitive inhibition can also be incorporated in the model. Experimental aspects of model calibration and verification are discussed. The need for models that distinguish between the exhaustion of cell activity and cell death is emphasized. © 1993 Wiley & Sons, Inc.
Journal Article•10.1002/BIT.260410707•
A statistical analysis of the effect of substrate utilization and shear stress on the kinetics of biofilm detachment

[...]

Brent M. Peyton1, Brent M. Peyton2, William G. Characklis1•
Montana State University1, Battelle Memorial Institute2
25 Mar 1993-Biotechnology and Bioengineering
TL;DR: The goal of this research was to determine the effects of shear stress and substrate loading rate on the rate of biofilm detachment.
Abstract: One of the least understood processes affecting biofilm accumulation is detachment. Detachment is the removal of cells and cell products from an established biofilm and subsequent entrainment in the bulk liquid. The goal of this research was to determine the effects of shear stress and substrate loading rate on the rate of biofilm detachment. Monopopulation Pseudomonas aeruginosa and undefined mixed population biofilms were grown on glucose in a RotoTorque biofilm reactor. Three levels of shear stress and substrate loading rate were used to determine their effects on the rate of detachment. Suspended cell concentrations were monitored to determine detachment rates, while other variables were measured to determine their influence on the detachment rate. Results indicate that detachment rate is directly related to biofilm growth rate and that factors which limit growth rate will also limit detachment rate. No significant influence of shear on detachment rate was observed. A new kinetic expression that incorporates substrate utilization rate, yield, and biofilm thickness was compared to published detachment expressions and gives a better correlation of data obtained both in this research and from previous research projects, for both mono- and mixed-population biofilms. © John Wiley & Sons, Inc.
Journal Article•10.1002/BIT.260410509•
Dramatic enhancement of enzymatic activity in organic solvents by lyoprotectants

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Katherine Debulis1, Alexander M. Klibanov1•
Massachusetts Institute of Technology1
05 Mar 1993-Biotechnology and Bioengineering
TL;DR: Activity enhancement induced by the excipients is displayed even after their removal by washing enzymes with anhydrous solvents, indicating that the mechanism of the excipient‐induced activation is based on the ability of theexcipients to alleviate reversible denaturation of enzymes upon lyophilization.
Abstract: When seven different hydrolytic enzymes (four proteases and three lipases) were lyophilized from aqueous solution containing a ligand, N-Ac-L-Phe-NH2, their catalytic activity in anhydrous solvents was far greater (one to two orders of magnitude) than that of the enzymes lyophilized without the ligand. This ligand-induced activation was expressed regardless of whether the substrate employed in organic solvents structurally resembled the ligand. Furthermore, nonligand lyoprotectants [sorbitol, other sugars, and poly(ethylene glycol)] also dramaticaliy enhanced enzymatic activity in anhydrous solvents when present in enzyme aqueous solution prior to lyophilization. The effects of the ligand and of the lyoprotectants were nonadditive, suggesting the same mechanism of action. Excipient activated and nonactivated enzymes exhibited identical activities in water. Also, addition of the excipients directly to suspensions of nonactivated enzymes in organic solvents had no appreciable effect on catalytic activity. These observations indicate that the mechanism of the excipient-induced activation is based on the ability of the excipients to alleviate reversible denaturation of enzymes upon lyophilization. Activity enhancement induced by the excipients is displayed even after their removal by washing enzymes with anhydrous solvents. Subtilisin Carlsberg, lyophilized with sorbitol, was found to be a much more efficient practical catalyst than its “regular” counterpart. © 1993 John Wiley & Sons, Inc.
Journal Article•10.1002/BIT.260410115•
A model of biofilm detachment

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Philip S. Stewart1•
Montana State University1
05 Jan 1993-Biotechnology and Bioengineering
TL;DR: A general mathematical framework for modeling biofilm detachment is presented, founded on a material balance on biomass that equates the detachment rate to the product of a detachment frequency and a detaching particle mass.
Abstract: A general mathematical framework for modeling biofilm detachment is presented. The approach is founded on a material balance on biomass that equates the detachment rate to the product of a detachment frequency and a detaching particle mass. The model provides a theoretical basis for deriving many of the empirical detachment rate expressions in common use and can thus lend some insight into their physical and biological significance. By allowing for variation in the detachment frequency with depth in the biofilm, the model permits derivation of detachment expressions that reflect a dependence on chemical or physiological gradients in the biofilm. Analysis of literature data sets from two different biofilm systems suggests, in both cases, that detachment is a growth-associated phenomenon. © 1993 John Wiley & Sons, Inc.
Journal Article•10.1002/BIT.260420508•
Substantial overproduction of antibodies by applying osmotic pressure and sodium butyrate.

[...]

Steve Oh1, Pooja Vig1, Florence Chua1, W.K. Teo1, Miranda G.S. Yap1 •
National University of Singapore1
20 Aug 1993-Biotechnology and Bioengineering
TL;DR: This study demonstrates that hybridomas can be hyperstimulated to produce higher immunoglobulin (lg) levels by suppressing cell growth and increasing culture longevity through adaptation to higher osmolarity media and addition of sodium butyrate.
Abstract: Much of the current cell technology has enabled increased antibody production levels due to judicious nutrient feeding to raise cell densities and design better bioreactors. This study demonstrates that hybridomas can be hyperstimulated to produce higher immunoglobulin (lg) levels by suppressing cell growth and increasing culture longevity through adaptation to higher osmolarity media and addition of sodium butyrate. Prior to adaptation, cells placed in higher osmotic pressures (350 and 400 mOsm) were severely suppressed in growth down to 25% of the control (300 mOsm), although total lg titers achieved were similar to the control, approximately 140 mg/L. After a week of adaptation to 350 and 400 mOsm media, cell growth was not as dramatically suppressed, but considerably higher lg levels were attained at these elevated osmolarities. The highest yield of 265 mg/L was obtained at 350 mOsm compared to 140 mg/L at 300 mOsm, while maximum viable cell numbers dropped from 35 x 10(5) cells/mL to 31 x 10(5) cells/mL and culture longevity was extended by 20 h more than the control. Sodium butyrate, known to enhance protein production in other cell types, was then supplemented at a range of concentrations between 0.01 and 0.4 mM to the 350 mOsm culture to further enhance the lg levels. Butyrate at a concentration of 0.1 mM, in combination with osmotic pressure at 350 mOsm, further elevated the lg levels to 350 mg/L. Concomitantly, maximum viable cell numbers were reduced to 22 x 10(5) cells/mL, but culture longevity was extended by 40 h in the 0.1 mM butyrate supplemented culture compared to the control condition. Specific antibody productivity, q(Mab), continued to stay high during the stationary phase and was further elevated during the decline phase: thus, overall lg levels can be increased by 2.3 times by combining osmotic pressure and butyrate treatment.
Journal Article•10.1002/BIT.260410302•
Industrial separation of carboxylic and amino acids by liquid membranes: applicability, process considerations, and potential advantage.

[...]

Aharon Eyal1, Eyal Bressler1•
Hebrew University of Jerusalem1
05 Feb 1993-Biotechnology and Bioengineering
TL;DR: Stability of the liquid advantages, can be secured by utilizing extractant blocking polymeric membranes, and economic implications for recovery for carboxylic and amino acids by various extractant/membrane combinations are discussed.
Abstract: Liquid–liquid extraction and membrane separation are well-known separation method of extensive industrial application. Their incorporation into liquid membranes has the potential of several advantages, some of which are of particular interest for the recovery of carboxylic and amino acids: selectivities higher than those attainable by current separation methods, saving on energy costs for final concentration of separated products, high fluxes, compact installation, and low capital and operation costs. Stability of the liquid advantages, can be secured by utilizing extractant blocking polymeric membranes, Applicability, process consideration, and economic implications for recovery for carboxylic and amino acids by various extractant/membrane combinations are discussed. © 1993 John Wiley & Sons, Inc.
Journal Article•10.1002/BIT.260420714•
A two-plane tubular photobioreactor for outdoor culture of Spirulina.

[...]

Giuseppe Torzillo1, Pietro Carlozzi1, Benjamin Pushparaj1, Elena Montaini1, R. Materassi1 •
University of Florence1
20 Sep 1993-Biotechnology and Bioengineering
TL;DR: Viscosity measurements performed on Spirulina cultures having different biomass concentrations showed non‐Newtonian behavior displaying decreasing viscosity with an increasing shear rate, which corresponded to a net photosynthetic efficiency of 6.6% (based on visible irradiance).
Abstract: A photobioreactor in the form of a 245-m-long loop made of plexiglass tubes having an inner diameter of 2.6 cm was designed and constructed for outdoor culture of Spirulina. The loop was arranged in two planes, with 15 8-m-long tubes in each plane. In the upper plane, the tubes were placed in the vacant space between the ones of the lower plane. The culture recycle was performed either with two airlifts, one per plane, or with two peristaltic pumps. The power required for water recycle in the tubular photobioreactor, with a Reynolds number of 4000, was 3.93 x 10(-2) W m(-2). The photobioreactor contained 145 L of culture and covered an overall area of 7.8 m(2). The photobioreactor operation was computer controlled. Viscosity measurements performed on Spirulina cultures having different biomass concentrations showed non-Newtonian behavior displaying decreasing viscosity with an increasing shear rate. The performance of the two-plane photobioreactor was tested under the climatic conditions of central Italy (latitude 43.8 degrees N, longitude 11.3 degrees E). A biomass concentration of 3.5 g L(-1) was found to be adequate for outdoor culture of Spirulina. With a biomass concentration of 6.3 g L(-1), the biomass output rate significantly decreased. The net biomass output rate reached a mean value of 27.8 g m(-2) d(-1) in July; this corresponded to a net photosynthetic efficiency of 6.6% (based on visible irradiance).
Journal Article•10.1002/BIT.260410202•
Improved activity retention of enzymes deposited on solid supports.

[...]

Ernst Wehtje1, Patrick Adlercreutz1, Bo Mattiasson1•
Lund University1
20 Jan 1993-Biotechnology and Bioengineering
TL;DR: The amount of additive needed for stabilization was shown to depend on the structure of the support, more additive being required for a support with high porosity, and the effect of the additives was studied using mandelonitrile lyase, but α‐chymotrypsin and lipase P were also shown to be stabilized.
Abstract: Enzymes deposited on solid support usually show good stability when operated in organic solvents. Decreased stability of the enzyme preparations was noticed when low enzyme loadings were used (e.g., with Celite as support; less than 1 mg enzyme/g). It was possible to avoid the activity loss by the addition of an additive which protects the enzyme during the immobilization. Proteins (such as albumin, gelatin, and casein) and poly(ethylene glycol) were effective additives whereas amino acids, monomeric carbohydrates, and polysaccharides had no effect. The amount of additive needed for stabilization was shown to depend on the structure of the support, more additive being required for a support with high porosity. The stabilizing effect was investigated in a series of glyceryl‐controlled‐pore glass (CPG) with varying specific surface areas (9.5–180 m2/g). The minimum addition of albumin, giving full stabilization, on the different supports correlated to a monolayer coverage of the surface, approximately 2–3 mg protein/m2. The effect of the additive was less pronounced when increasing amounts of enzyme were immobilized (5–40 mg enzyme/g Celite). The effect of the additives was studied using mandelonitrile lyase, but α‐chymotrypsin and lipase P were also shown to be stabilized. © 1993 John Wiley & Sons, Inc.
Journal Article•10.1002/BIT.260410703•
Phenol removal from waste gases with a biological filter by Pseudomonas putida

[...]

Mario Zilli, Attilio Converti, Alessandra Lodi, M. Del Borghi, G. Ferraiolo 
25 Mar 1993-Biotechnology and Bioengineering
TL;DR: Investigation of the feasibility of biologically removing phenol from waste gases by means of a biofilter using a Pseudomonas putida strain showed that the pure culture is more efficient than the mixed one, even when the latter has undergone three successive acclimatization tests.
Abstract: The purpose of this study is to investigate the feasibility of biologically removing phenol from waste gases by means of a biofilter using a Pseudomonas putida strain. Two series of both batch and continuous tests have been performed in order to ascertain the microbial degradation of phenol. For the preliminary batch tests, carried out in order to test the effective feasibility of the process and to investigate their kinetic behavior, two different microbial cultures belonging to the Pseudomonas genus have been employed, a heterogeneous culture and a pure strain of P. putida. The results of these comparative investigation showed that the pure culture is more efficient than the mixed one, even when the latter has undergone three successive acclimatization tests. The continuous experiments have been conducted during a period of about 1 year in a laboratory-scale column, packed with a mixture of peat and glass beads, and utilizing the pure culture of P. putida as microflora and varying the inlet phenol concentration from 50 up to 2000 mg m(-3). The results obtained show that high degrees of conversion can be obtained (0.93/0.996) operating at a residence time of 54 s.
Journal Article•10.1002/BIT.260411002•
A novel membrane bioreactor for detoxifying industrial wastewater: I. Biodegradation of phenol in a synthetically concocted wastewater

[...]

Andrew G. Livingston1•
Imperial College London1
25 Apr 1993-Biotechnology and Bioengineering
TL;DR: A mathematical model showed that the attached biofilm significantly lowers the mass transfer driving force for phenol across the membrane, and oxygen concentration limits the phenol degradation rate in the biofilm.
Abstract: A novel process has been used to biodegrade phenol present in an acidic (1 M HCI) and salty (5% w/w NaCl) synthetically bioreactor, in which the phenol present in the wastewater is separated from the inorganic components by means of a silicone rubber membrane. Transfer of the phenol from the wastewater and into a biological growth medium allows biodegradation to proceed under controlled conditions which are unaffected by the hostile inorganic composition of the wastewater. At a wastewater flow rate of 18 mL h(-1) (contact time 6 h), 98.5% of the phenol present in the wastewater at an inlet concentration of 1000 mg ((-1)) was degraded; at a contact time of 1.9 h, 65% of the phenol was degraded. Phenol degradation was accompanied by growth of a biofilm on the membrane tubes and by conversion of approximately 80% of the carbon entering the system to CO(2) carbon. Analysis of the transport of phenol across the membrane revealed that the major resistance to mass transfer arose in the diffusion of phenol across the silicone rubber membrane. A mathematical model was used to describe the transfer of phenol across the membrane and the subsequent diffusion and reaction of phenol in the biofilm attached to the membrane tube. This analysis showed that (a) the attached biofilm significantly lowers the mass transfer driving force for phenol across the membrane, and (b) oxygen concentration limits the phenol degradation rate in the biofilm. These conclusions from the model are consistent with the experimental results.
Journal Article•10.1002/BIT.260420710•
Removal of phenols from wastewater by soluble and immobilized tyrosinase

[...]

Wada Shinji, Hiroyasu Ichikawa, Kenji Tatsumi
20 Sep 1993-Biotechnology and Bioengineering
TL;DR: An enzymatic method for removal of phenols from industrial wastewater was investigated and immobilized tyrosinase, immobilized by using amino groups in the enzyme on cation exchange resins, can be used repeatedly.
Abstract: An enzymatic method for removal of phenols from industrial wastewater was investigated. Phenols in an aqueous solution were removed after treatment with mushroom tyrosinase. The reduction order of substituted phenols is catechol > p-cresol > p-chlorophenol > phenol > p-methoxyphenol. In the treatment of tyrosinase alone, no precipitate was formed but a color change from colorless to dark-brown was observed. The colored products were removed by chitin and chitosan which are available abundantly as shellfish waste. In addition, the reduction rate of phenols was observed to be accelerated in the presence of chitosan. Tyrosinase, immobilized by using amino groups in the enzyme on cation exchange resins, can be used repeatedly. By treatment with immobilized tyrosinase, 100% of phenol was removed after 2 h, and the activity was reduced very little even after 10 repeat treatments.
Journal Article•10.1002/BIT.260410804•
Recombinant trypsin production in high cell density fed-batch cultures in Escherichia coli

[...]

L. Yee1, Harvey W. Blanch1•
University of California, Berkeley1
05 Apr 1993-Biotechnology and Bioengineering
TL;DR: The induction timing is shown to have a significant effect on the final cell density as well as the overall volumetric yield of the recombinant protease.
Abstract: Fed-batch techniques were employed to obtain high cell density cultures (92-100 g DCW/L) of Escherichia coli strain X90 producing a recombinant serine protease, rat anionic trypsin, secreted to the periplasm. The specific growth rate was controlled to minimize growth-inhibiting acetate formation by utilizing an exponential feeding profile determined from mass balance equation. The volumetric yield of recombinant rat anionic trypsin was 56 mg/L, and the final cell density was 92 g DCW/L when the culture was induced in the late logarithmic phase. However, when the culture was induced in the early logarithmic phase, the volumetric yield was 13 mg/L and the final cell density was 14 g DCW/L. Thus, the induction timing is shown to have a significant effect on the final cell density as well as the overall volumetric yield of the recombinant protease.
Journal Article•10.1002/BIT.260420616•
Lipase-catalyzed acyiation of sugars solubilized in hydrophobic solvents by complexation.

[...]

Isao Ikeda1, Alexander M. Klibanov1•
Massachusetts Institute of Technology1
05 Sep 1993-Biotechnology and Bioengineering
TL;DR: By complexing glucose and other mono- and disaccharides with phenylboronic acid, they were solubilized in many organic solvents and produced a sugar-based polyacrylate with a molecular weight of 14,000 Da.
Abstract: By complexing glucose and other mono and disaccharides with phenylboronic acid, they were solubilized in many organic solvents. Pseudomonas sp. lipoprotein lipase readily acylated such solubilized sugars in these solvents (no reaction was observed without phenylboronic acid, i.e., when sugars are insoluble in the reaction medium). Solubilized D-glucose was enzymatically acylated with vinyl acrylate on a preparative scale, and the resultant 6-O-acryloylglucose was chemically polymerized to yield a sugar-based polyacrylate with a molecular weight of 14,000 Da.
Journal Article•10.1002/BIT.260410507•
High cell density cultivation of Pseudomonas oleovorans: Growth and production of poly (3‐hydroxyalkanoates) in two‐liquid phase batch and fed‐batch systems

[...]

Hans Preusting1, Renze Van-Houten1, Andre Hoefs1, Eddy-Kool Van-Langenberghe1, Olivier Favre-Bulle1, Bernard Witholt1 •
University of Groningen1
05 Mar 1993-Biotechnology and Bioengineering
TL;DR: This study aimed at an efficient production of poly(3‐hydroxyalkanoates) by growing P. oleovorans to high cell densities in fed‐batch cultures.
Abstract: Pseudomonas oleovorans is able to accumulate poly(3-hydroxyalkanoates) (PHAs) under conditions of excess n-alkanes, which serve as sole energy and carbon source, and limitation of an essential nutrient such as ammonium. In this study we aimed at an efficient production of these PHAs by growing P. oleovorans to high cell densities in fed-batch cultures.To examine the efficiency of our reactor system, P. oleovorans was first grown in batch cultures using n-octane as growth substrate and ammonia water for pH regulation to prevent ammonium limiting conditions. When cell growth ceased due to oxygen limiting conditions, a maximum cell density of 27 g .L(-1) dry weight was obtained. When the growth temperature was decreased from the optimal temperature of 30 degrees -18 degrees C, cell growth continued to a final cell density of 35 g . L(-1) due to a lower oxygen demand of the cells at this lower incubation temperature.To quantify mass transfer rates in our reactor system, the volumetric oxygen transfer coefficient (k(L)a) was determined during growth of P. oleovorans on n-octane. Since the stirrer speed and airflow were increased during growth of the organism, the k(L)a also increased, reaching a constant value of 0.49 s(-1) at maximum airflow and stirrer speed of 2 L . min(-1) and 2500 rpm, respectively. This k(L)a value suggests that oxygen transfer is very efficient in our stirred tank reactor.Using these conditions of high oxygen transfer rates, PHA production by P. oleovorans in fed-batch cultures was studied. The cells were first grown batchwise to a density of 6 g . L(-1), after which a nutrient feed, consisting of (NH(4))(2)SO(4) and MgSO(4), was started. The limiting nutrient ammonium was added at a constant rate of 0.23 g NH(4) (+) per hour, and when after 38 h the feed was stopped, a biomass concentration of 37.1 g . L(-1) was obtained. The Cellular PHA content was 33% (w/w), which is equal to a final PHA yield of 12.1 g . L(-1) and an overall PHA productivity of 0.25 g PHA produced per liter medium per hour.
Journal Article•10.1002/BIT.260410710•
Diffusivity of Cu2+ in calcium alginate gel beads

[...]

Dong Chen1, Zbigniew Lewandowski1, Frank L. Roe1, Prasad Surapaneni1•
Montana State University1
25 Mar 1993-Biotechnology and Bioengineering
TL;DR: A linear absorption model (LAM) is used to describe the process of metal binding to spherically shaped biopolymers particles and has theoretical advantages over the shrinking core model (shell progressive model), which calculated an unreasonable exponential increase in the diffusion coefficient as density of alginate polymer in the bead increased.
Abstract: A linear absorption model (LAM) is used to describe the process of metal binding to spherically shaped biopolymers particles. The LAM was solved using a numerical algorithm which calculates diffusivities of metal ion in biopolymer gels. It assumes attainment of rapid metal-biopolymer binding equilibrium accompanied by rate limiting diffusion of the metal ions through the gel. The model was tested using batch experiments in which copper (Cu(2+)) binding with calcium alginate beads was investigated. Biopolymer density in the beads was varied between 2% and 5%. The diffusion coefficient of Cu(2+) calculated from the LAM ranged from 1.19 x 10(-9) to 1.48 x 10(-9) m(2) s(-1) (average 1.31 +/- 0.21 x 10(-9) m(2) s(-1)), independent of biopolymer density. The LAM has theoretical advantages over the shrinking core model (shell progressive model). The latter calculated an unreasonable exponential increase in the diffusion coefficient as density of alginate polymer in the bead increased.
Journal Article•10.1002/BIT.260420419•
Microbial utilization of levoglucosan in wood pyrolysate as a carbon and energy source.

[...]

Elizabeth Prosen1, D. Radlein1, Jan Piskorz1, Donald S. Scott1, Raymond L. Legge1 •
University of Waterloo1
05 Aug 1993-Biotechnology and Bioengineering
TL;DR: It is apparent that a wood pyrolysate maximized for levoglucosan can serve as a fermentable substrate, although postpyrolysis clean-up appears necessary.
Abstract: The Waterloo Fast Pyrolysis Process (WFPP) can produce an organic liquid high in levoglucosan (1, 6-anhydro-beta-D-glucopyranose) content from suitably pretreated lignocellulosics. A variety of fungi and yeasts were screened for their ability to utilize and ferment this organic liquid. To enhance its fermentability, the pyrolysis tar was posttreated in three different ways: (1) an aqueous extract (lignin removed); (2) activated charcoal treated (lignin and aromatics removed); and (3) acid hydrolysate (lignin and aromatics removed with the levoglucosan hydrolyzed to glucose). Four fungal strains were examined. None grew in the aqueous extract, but all grew equally well in both the activated charcoal treated and the acid hydrolysate, suggesting that the aromatic species were inhibitory to growth. Seven yeast species were examined, two of which did not grow on any of the extracts. Five of the yeast strains grew well on both the aqueous extract as well as the activated charcoal extract. The hydrolysate was optimal in terms of biomass yield and ethanol production. Ethanol yields on the hydrolysate were comparable or better than those on glucose. Ethanol was also produced in the aqueous extract and activated charcoal-treated substrate, but yields were considerably lower than on the hydrolysate or glucose. It is apparent that a wood pyrolysate maximized for levoglucosan can serve as a fermentable substrate, although postpyrolysis clean-up appears necessary.
Journal Article•10.1002/BIT.260410907•
Purification and characterization of a highly thermostable glucose isomerase produced by the extremely thermophilic eubacterium, Thermotoga maritima.

[...]

Stephen H. Brown1, Stephen H. Brown2, Carsten Sjøholm3, Robert M. Kelly1, Robert M. Kelly2 •
University of Maryland, Baltimore1, Johns Hopkins University2, Novo Nordisk3
15 Apr 1993-Biotechnology and Bioengineering
TL;DR: The high degree of thermostability, coupled with a neutral to slightly acid pH optimum, reveal this enzyme to be a promising candidate for improvement of the industrial glucose isomerization process.
Abstract: Thermotoga maritima, among the most thermophilic eubacteria currently known, produces glucose isomerase when grow in the presence of xylose. The purified enzyme is a homotetramer with submit molecular Wight of about 45,000. It has a number of features in common with previously described glucose isomerases-pH optimum of 6.5 to 7.5, presence of active-site histidine, requirement for metal cations such as Co(2+) and Mg(2+), and preference for xylose as substrate. In addition, it has significant sequence/structural homology with other glucose isomerases, as shown by both N-terminal sequencing and immunological crossreactivity. The T. maritima enzyme is distinguished by its extreme thermostability-a temperature optimum of 105 to 110 degrees C, and an estimated half-life of 10 minutes at 120 degrees C, pH 7.0. The high degree of thermostability, coupled with a neutral to slightly acid pH optimum, reveal this enzyme to be a promising candidate for improvement of the industrial glucose isomerization process.
Journal Article•10.1002/BIT.260410706•
A simple morphologically structured model describing the growth of filamentous microorganisms.

[...]

Jens Nielsen1•
Technical University of Denmark1
25 Mar 1993-Biotechnology and Bioengineering
TL;DR: A simple morphologically structured growth model is set up that may describe the growth of filamentous microorganisms both on a solid medium and in a submerged culture.
Abstract: Based on the reported mechanisms for filamentous growth, a simple morphologically structured growth model is set up. The model may describe the growth of filamentous microorganisms both on a solid medium and in a submerged culture. For description of a submerged culture the model is combined with a simple population model, which is derived from a balance for the distribution function for the hyphal elements. The model is compared with experimental data for three species of filamentous microorganisms: Geotrichum candidum, Streptomyces hygroscopicus, and Penicillium chrysogenum. © 1993 John Wiley & Sons, Inc.
Journal Article•10.1002/BIT.260420103•
Viability testing and characterization of germination of fungal spores by automatic image analysis.

[...]

G. C. Paul1, C.A. Kent1, Colin R. Thomas1•
University of Birmingham1
05 Jun 1993-Biotechnology and Bioengineering
TL;DR: An image analysis method has been developed for assessing the viability and the germination characteristics of fungal spores in submerged cultures and might be used as a tool in culture maintenance and for determining the quality of inocula for fungal fermentations.
Abstract: Fungal spores are used in the laboratory for culture maintenance and at laboratory and other scales as inocula for fermentations. The spore swelling and germination processes constitute a major part of the lag phase, and the subsequent culture morphology and productivity can be greatly influenced by the initial concentration and condition of the spores. An image analysis method has been developed for assessing the viability and the germination characteristics of fungal spores in submerged cultures. Structural variations during germination, i.e., swelling, germ tube formation, and germ tube elongation, are measured in terms of distributions of spore volumes and of germ tube lengths and volumes. These measurements are fully automatic and give a very rapid assessment of spore viability. This image analysis method might be used as a tool in culture maintenance and for determining the quality of inocula for fungal fermentations.
...

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