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Showing papers in "Biotechnology and Bioengineering in 1979"


Journal ArticleDOI
TL;DR: Inhibitory effects of ethanol on the yeast strain Saccharomyces cerevisiae NRRL‐Y‐132 were studied in batch and continuous chemostat cultures and the degree of substrate inhibition was higher than that reported for ethanol fermentation of pure sugar.
Abstract: High concentrations of both ethanol and sugar in the fermentation broth inhibit the growth of yeast cells and the rate of product formation. Inhibitory effects of ethanol on the yeast strain Saccharomyces cerevisiae NRRL‐Y‐132 were studied in batch and continuous chemostat cultures. Growth was limited by either glucose or ethanol. Feed medium was supplemented with different ethanol concentrations. Ethanol was found to inhibit growth and the activity of yeast to produce ethanol in a noncompetitive manner. A linear kinetic pattern for growth and product formation was observed according to μ = μm (1 – P/Pm) and v = vm (1 – P/Pm′), where μm is the maximum specific growth rate at P = 0 (hr−1); Pm is the maximum specific product formation rate at P = 0 (hr−1); Pm is the maximum ethanol concentration above which cells do not grow (g/liter); Pm′ is the maximum ethanol concentration above which cells do not produce ethanol (g/liter). Substrate inhibition studies were carried out using short‐time experimental techniques under aerobic and anaerobic condition. The degree of substrate inhibition was found to be higher than that has been reported for ethanol fermentation of pure sugar. The kinetic relationships thus obtained were used to compute growth, substrate utilization, and alcohol production patterns and have been discussed with reference to batch and continuous fermentation of enzymatically produced bagasse hydrolysate.

265 citations


Journal ArticleDOI
TL;DR: Columns of calcium alginate gel pellets have excellent physical properties when used as a cell immobilization support and can be chosen so as to form strong pellets, possessing high substrate transfer rates and low rates of cell leakage.
Abstract: Columns of calcium alginate gel pellets have excellent physical properties when used as a cell immobilization support. Columns of pellets were very resistant to compression and abrasion during passage of high concentrations of sucrose at high flow rates, but if the pellets were formed using low alginate and Ca2+ concentrations, compression occurred and flow out of the column was reduced and pressure built up. Transfer of sucrose into the pellets was controlled by internal diffusion, the rate of diffusion being increased by reductions in the alginate and Ca2+ concentrations used for immobilization and by the presence of entrapped active cells. Some leakage of cells occurred during use especially when cell division of the entrapped cells took place, but leakage could be minimized by using more highly polymerized pellets. Therefore, immobilization conditions can be chosen so as to form strong pellets, possessing high substrate transfer rates and low rates of cell leakage.

232 citations


Journal ArticleDOI
TL;DR: In this article, a yeast strain was used to convert bagasse hydrolysate to ethanol under anaerobic conditions by a strain of Saccharomyces cerevisiae in batch and continuous cultures.
Abstract: Rapid fermentation of bagasse hydrolysate to ethanol under anaerobic conditions by a strain of Saccharomyces cerevisiae has been studied in batch and continuous cultures at pH 4.0 and 30°C temperature with cell recycle. By using a 23.6 g/liter cell concentration, a concentation of 9.7% (w/v)ethanol was developed in a period of 6 hr. The rate of fermentation was found to increase with supplementation of yeast vitamins in the hydrolysate. In continuous culture employing cell recycle and a 0.127 v/v/m air flow rate, a cell mass concentration of 48.5 g/liter has been achieved. The maximum fermentor productivity of ethanol obtained under these conditions was 32.0 g/liter/hr, which is nearly 7.5 times higher than the normal continuous process without cell recycle and air sparging. The ethanol productivity was found to decrease linearly with ethanol concentration. Conversion of glucose in the hydrolysate to ethanol was achieved with a yield of 95 to 97% of theoretical.

228 citations


Journal ArticleDOI
TL;DR: Simultaneous adsorption of exo‐ and endoglucanases on hydrolyzable cellulosics is the causative factor of the hydrolysis that follows immediately after, and supports the postulate of synergistic enzyme action proposed by Eriksson.
Abstract: Most cellulosic substances contain appreciable amounts of cellulose and hemicellulose, which on enzymatic hydrolysis mainly yield a mixture of glucose, cellobiose, and xylose. In this paper, studies on the mechanisms of hydrolysis of bagasse (a complex native cellulosic waste left after extraction of juice from cane sugar) by the cellulase enzyme components are described in light of their adsorption characteristics. Simultaneous adsorption of exo- and endoglucanases on hydrolyzable cellulosics is the causative factor of the hydrolysis that follows immediately after. It supports the postulate of synergistic enzyme action proposed by Eriksson. Xylanase pretreatment enhanced the hydrolysis of bagasse owing to the creation of more accessible cellulosic regions that are readily acted upon by exo- and endoglucanases. The synergistic action of the purified exoglucanase, endoglucanase, and xylanse has been found to be most effective for hydrolysis of bagasse but not for pure cellulose. Significant quantities of glucose are produced in beta-glucosidase-free cellulase action on bagasse. Individual and combined action of the purified cellulase components on hydrolysis of native and delignified bagasse are discussed in respect to the release of sugars in the hydrolysate.

183 citations


Journal ArticleDOI
TL;DR: Results are presented to show the effectiveness of simultaneous anticipatory and feedback contol in overcoming problems of oxygen starvation, molasses quality, and variable inoculum size.
Abstract: The Biotechnology and Bioengineering study presented here was undertaken to demonstrate the usefulness of computer control for the production of yeast from molasses. A flexible control system was developed by using an on-line computer for the monitoring of cell mass and employing anticipatory control to maintain the maximum productivity. Process disturbances were minimized by employing a multivariable feedback control system to prevent ethanol formation. The control strategy acted to keep overall conversion yield at its maximum level, about 0.5 g cell/ g sugar, while maintaining high volumetric productivity between 3 and 5 g/liter-hr. Results are presented to show the effectiveness of simultaneous anticipatory and feedback contol in overcoming problems of oxygen starvation, molasses quality, and variable inoculum size.

156 citations


Journal ArticleDOI
TL;DR: Conditions for the gelation of k ‐carrageenan, which is a new polymer for immobilization of enzymes and microbial cells, were investigated in detail and immobilized preparations were easily tailor‐made to various shapes such as cube, bead, and membrane.
Abstract: Conditions for the gelation k-carrageenan, which is a new polymer for immobilization of enzymes and microbial cells, were investigated in detail k-Carrageenan was easily induced to gel by contact with metal ions, amines, amino acid derivatives, and water-miscible organic solvents By using this property of k-carrageenan, the immobilization of enzymes and microbial cells was investigated Several kinds of enzymes and microbial cells were easily immobilized with high enzyme activities Immobilized preparations were easily tailor-made to various shape such as cube, bead, and membrane The obtained immobilized preparations were stable, and columns packed with them were used for continuous enzyme reaction for a long period Their operational stabilities were enhanced by hardening with glutaraldehyde and hexamethylenediamine

154 citations


Journal ArticleDOI
TL;DR: The hypothesis that bead adsorption of nutrients is responsible for such “toxicities” associated with this technique was found to be inconsistent with results, which suggest that microenvironmental effects are critical for cell propagation on microcarriers.
Abstract: The microcarrier technique for the growth of anchorage‐dependent animal cells has been studied and significantly improved. Excellent cell growth (up to 5 × 106 cells/ml) has been obtained on a newly synthesized microcarrier optimized with respect to substitution with a positively charged exchange moiety. Various parameters of microcarrier culture were examined in order to identify the source of reported “toxicities” associated with this technique. The hypothesis that bead adsorption of nutrients is responsible for such “toxicities” was found to be inconsistent with our results, which suggest that microenvironmental effects are critical for cell propagation on microcarriers.

146 citations


Journal ArticleDOI
TL;DR: In this paper, a novel biomass support particle containing growing cells was developed for use in large-scale fermentation processes, which can be produced of any size, shape, and density with a wide variety of microorganisms.
Abstract: Novel biomass support particles containing growing cells have been developed for use in large-scale fermentation processes. The characteristic size of the entrapped biomass is identical to that of the physical structure of the support particle, and particles can be produced of any size, shape, and density with a wide variety of microorganisms. Use of the particles in fermentors leads to high biomass concentration independent of throughput, predetermined biomass concentrations, the use of novel types of fermentor with advantageous performance characteristics, possibilities for the optimization of advantageous diffusion effects, and new procedures for biomass recovery.

128 citations


Journal ArticleDOI
TL;DR: It is shown that the feed rate profile during fermentation is of vital importance in the realization of a high production rate throughout the duration of the fermentation, and the method of modeling presented may also prove rewarding for an analysis of fermentation processes other than the penicillin fermentation.
Abstract: This paper shows the application of elementary balancing methods in combination with simple kinetic equations in the formulation of an unstructured model for the fed-batch process for the production of penicillin. The rate of substrate uptake is modeled with a Monod-type relationship. The specific penicillin production rate is assumed to be a function of growth rate. Hydrolysis of penicillin to penicilloic acid is assumed to be first order in penicillin. In simulations with the present model it is shown that the model, although assuming a strict relationship between specific growth rate and penicillin productivity, allows for the commonly observed lag phase in the penicillin concentration curve and the apparent separation between growth and production phase (idiophase-trophophase concept). Furthermore it is shown that the feed rate profile during fermentation is of vital importance in the realization of a high production rate throughout the duration of the fermentation. It is emphasized that the method of modeling presented may also prove rewarding for an analysis of fermentation processes other than the penicillin fermentation.

122 citations


Journal ArticleDOI
TL;DR: The concept of mass balance was used to analyze the metabolic pathways of citrate production by Candida lipolytica from glucose, and models I, II, and III were found to be most unlikely because of the unusual flux assessed from these models.
Abstract: The concept of mass balance was used to analyze the metabolic pathways of citrate production by Candida lipolytica from glucose. Specific rates of glucose consumption, citrate and isocitrate productions, carbon dioxide evolution, and cellular syntheses of protein and carbohydrate were observed in an NH4+-limited chemostat culture. These data permitted one to assess the carbon flux in vivo by solving simultaneous carbon balance equations with respect to intermediary metabolite pools in the steady State. Among the three models considered here, model I (which coordinates the pyruvate carboxylation with the tricarboxylic acid cycle, but disregards the glyoxylate cycle) was considered plausible because the carbon flux calculated so far was acceptable. On the other hand, models II and III (which overlook the pyruvate carboxylation and the 2-oxoglutarate dehydrogenation, respectively) were found to be most unlikely because of the unusual flux assessed from these models.

113 citations


Journal ArticleDOI
TL;DR: Cellulase synthesis was strongly repressed in the presence of glucose and only a low constitutive activity of β‐glucosidase and carboxymethylcellulase, but no Avicelase, could be demonstrated when culturing T. viride on glucose.
Abstract: Trichoderma viride ITCC-1433 produces high yields of cellulase and especially β-glucosidase when grown in submerged culture on different carbon sources. Cellulase synthesis was strongly repressed in the presence of glucose and only a low constitutive activity of β-glucosidase and carboxymethylcellulase, but no Avicelase, could be demonstrated when culturing T. viride on glucose. With carboxymethylcellulose (CMC) as a substrate the secretion of enzyme as well as growth depended on the degree of substitution, but in general CMC cannot be regarded either as a powerful inducer or as a carbon source. With insoluble cellulose, maximum enzyme production and activities were obtained using an alkali-treated cellulose powder. On this substrate the excretion of soluble protein into the culture broth increased and the protein concentration corresponded to cellulolytic activities.

Journal ArticleDOI
TL;DR: Shear forces alone were not found to be as effective in causing enzyme inactivation as is generally believed and alternative mechanisms for damage are discussed.
Abstract: Intermittent shear was applied to approximately 1 mg/ml solutions of bovine liver catalase in a coaxial cylindrical viscometer at temperatures from 20 to 60°C and shear rates up to 683 sec−1. The viscometer was sealed to prevent evaporation. Up to 40°C there were no activity losses during 3 hr total shearing. Above 40°C shearing reduced losses due to thermal inactivation, possibly by interfering with precipitation. At 3440 sec−1 and 40°C fine precipitates formed but little activity was lost. No activity losses were found with experimental conditions under which Taylor vortexing occurred, nor when shear stresses were increased up 57 times by adding glycerol to raise the, viscosity. There were no significant losses in a capillary rheometer at shear rates up to 106 sec−1. When low concentration (6 μg/ml) catalase solutions were sheared there was little loss in sealed systems, but there were losses in “open” systems even in low‐temperature nonshear experiments. Although there were no losses with 1 mg/ml solutions, 6 μg/ml catalase solutions from an alternative source did lose activity in sealed systems but much less than expected from previously published work. Approximately 1 mg/ml jack bean urease solutions were sheared in the sealed system at 23°C and 683 sec−1 for 3 hr. No losses were found. No evidence of temporary or permanent inactivation was found with 28°g/ml solutions sheared in the presence of urea. Shear forces alone were not found to be as effective in causing enzyme inactivation as is generally believed and alternative mechanisms for damage are discussed.

Journal ArticleDOI
TL;DR: Citric acid concentration increased rapidly as the dilution rate decreased and appears to be critically dependent on the pH in the culture vessel and the nitrogen concentration in the feed.
Abstract: Work has been carried out on the production of citric acid by Aspergillus foetidus in single‐stage continuous culture, operated under nitrogen‐limiting conditions at dilution rates between 0.04 to 0.21 hr−1. Citric acid concentration increased rapidly as the dilution rate decreased and appears to be critically dependent on the pH in the culture vessel and the nitrogen concentration in the feed. A mathematical model based on a distinction between basic cells, which require nitrogen but do not produce citric acid, and stroage cells, which accumulate carbon and simultaneously produce citric acid, is proposed.

Journal ArticleDOI
TL;DR: Yeast alcohol dehydrogenase (ADH) solutions were sheared in a coaxial cylindrical viscometer and it was shown that there were no ADH aggregates in the supernatant and that the precipitate was less than 2% of the original protein.
Abstract: Yeast alcohol dehydrogenase (ADH) solutions (approximately 1 mg/ml, pH 7) were sheared in a coaxial cylindrical viscometer. This was fitted with a lid sealing the contents from the atmosphere and preventing evaporation. At 30 degrees C after a total of 5 hr intermittent shearing at 683 sec-1 no losses of activity were observed. No losses were found after 5 hr continuous shearing and in a no-shear control. At 40 degrees C and 683 sec-1 there were only small activity losses in 5 hr. Shearing at 3440 sec-1 no measurable losses of activity were found with a 1.03 mg/ml solution in 5 hr at 30 degrees C, a 1.03 mg/ml solution in 8 hr at 5 degrees C, and with a 3.89 mg/ml solution in 3 hr at 5 degrees C. In all these cases, however, a white precipitate formed that was not observed in zero shear control experiments. The sheared 3.89 mg/ml solution was clarified by centrifugation. It was shown that there were no ADH aggregates in the supernatant and that the precipitate was less than 2% of the original protein. At 30 degrees C under adverse pH conditions (pH 8.8) there was no significant difference in activity losses of an approximately 1 mg/ml solution sheared at 65 and 744 sec-1. An approximately 0.5 mg/ml ADH solution, pH 7, was agitated in a small reactor with no free air-liquid interface. Peak shear rates near the impeller were estimated to be about 9000 sec-1. Only a small decrease in specific activity was observed until over 15 hr total running at 5 degrees C.

Journal ArticleDOI
TL;DR: The relationship between the degree of crystallinity of cellulose and its susceptibility to cellulase was investigated in this article, where the x-ray diffraction of pure cellulose was performed.
Abstract: Chemical and physical treatments of cotton cellulose have been studied in order to elucidate the relationship between the degree of crystallinity of cellulose and the susceptibility of cellulose to cellulase. Cotton cellulose powder was treated with the following solvents: 60% H/sub 2/SO/sub 4/, Cadoxen, and DMSO-P-formaldehyde. The dissolved celluloses were recovered at high yield of over 97% by addition of nine volumes of cold acetone. X-ray diffraction for measurements of relative crystallinity showed that the crystalline structure of cellulose declined in quantity and perfection by the dissolving treatment and changed to an amorphous form that is highly susceptible to enzymatic hydrolysis. These reprecipitated celluloses were hydrolyzed almost completely within 48 hours by Aspergillus niger cellulase containing mainly 1,4-Beta-glucan glucanohydrolase without action of 1,4-beta-glucan cellobiohydrolase. On the other hand, cryo-milled cellulose (below 250 mesh) still had a crystalline structure, was resistant to cellulase, and gave a low percentage of saccharification. These results indicate that in pure cellulose there are good correlations between x-ray diffractograms and susceptibility to microbial cellulase.

Journal ArticleDOI
TL;DR: In this article, a Teflon membrane and an oxygen electrode were used for the continuous determination of methyl and ethyl alcohols, and the current of the electrode decreased with time until a steady state was reached.
Abstract: A microbial electrode consisting of immobilized microorganisms, a gas permeable Teflon membrane, and an oxygen electrode was prepared for the continuous determination of methyl and ethyl alcohols. Immobilized Trichosporon brassicae was employed for a microbial electrode sensor for ethyl alcohol. When a sample solution containing ethyl alcohol was injected into a microbial electrode system, the current of the electrode decreased markedly with time until a steady state was reached. The response time was within 10 min by the steady state method and within 6 min by the pulse method. A linear relationship was observed between the current decrease and the concentration of ethyl alcohol below 22.5 mg/liter. The current was reproducible within ± 6% of the relative error when a sample solution containing 16.5 mg/liter ethyl alcohol. The standard deviation was 0.5 mg/liter in 40 experiments. The selectivity of the microbial electrode sensor for ethyl alcohol was satisfactory. The microbial electrode sensor was applied to a fermentation broth of yeasts and satisfactory comparative results were obtained (correlation coefficient 0.98). The current output of the microbial electrode sensor was almost constant for more than three weeks and 2100 assays. A microbial electrode sensor using immobilized bacteria for methyl alcohol was also described.

Journal ArticleDOI
TL;DR: For certain mixing conditions the maximum amount of releasable protein was dependent on the impeller geometry, construction material, and also the concentration of packed yeast, and the relative power efficiencies of the two mills are discussed along with possible criteria for scaling of bead mills.
Abstract: Laboratory and pilot-plant high-speed bead mills of 0.6 and 5 liter capacity and consisting of four and five impellers in series, respectively, were used to follow the batch and continuous disruption of bakers' yeast (Saccharomyces cerevisiae). The mills are not scaled equivalents. Throughputs ranging from 1 × 10−6m3/sec to 12 × 10−6m3/sec for the 0.6 liter mill and from 16 × 10−6m3/sec to 100 × 10−6m3/sec for the 5 liter mill were used for continuous disruption studies. Variables studied included the effect of impeller tip speed, temperature, and packed yeast concentration (ranging from 15 to75% by weight packed yeast). Disruption kinetics, as measured by the release of soluble protein, followed a first-order rate equation, the rate constant being a function of impeller tip speed and yeast concentration. For continuous disruption studies the bead mills behaved as a series of continuous stirred-tank reactors, each impeller forming a reactor. In the smaller mill a considerable degree of backflow between the reactors was evident. For certain mixing conditions the maximum amount of releasable protein was dependent on the impeller geometry, construction material, and also the concentration of packed yeast. The relative power efficiencies of the two mills are discussed along with possible criteria for scaling of bead mills.

Journal ArticleDOI
TL;DR: Material and energy balances for continuous‐culture processes are described based on the facts that the heat of reaction per electron transferred to oxygen for a wide variety of organic molecules, the number of available electrons per carbon atom in biomass, and the weight fraction carbon in biomass are relatively constant.
Abstract: Material and energy balances for continuous-culture processes are described based on the facts that the heat of reaction per electron transferred to oxygen for a wide variety of organic molecules, the number of available electrons per carbon atom in biomass, and the weight fraction carbon in biomass are relatively constant. Energy requirements for growth and maintenance are investigated and related to the biomass energetic yield. The consistency of experimental data is examined using material and energy balances and the regularities identified above. When extracellular products are absent, the consistency of yield models containing separate terms for growth and maintenance may be investigated using organic substrate consumption, biomass production, oxygen consumption (or heat evolution), and carbon dioxide evolution rate data for a series of dilution rates. The consistency of continuous-culture data in the published literature is examined.

Journal ArticleDOI
TL;DR: Trichoderma can be cultured in stirred‐tank fermentors on high (8%)cellulose concentrations without increasing the salt concentration of the medium when NH4OH is used to control pH and as a nitrogen source.
Abstract: Trichoderma can be cultured in stirred-tank fermentors on high (8%)cellulose concentrations without increasing the salt concentration of the medium when NH4OH is used to control pH and as a nitrogen source. Approximation 90% of the ammonia consumed by the organism can be added as NH4OH. The advantage of using high concentrations of cellulose is that culture filtrates with greater cellulase activity are obtained. The advantage of a low salts medium is that unwanted solutes in the final enzyme preparation are reduced. The appearance of cellulase in the medium occurs later than net ammonia uptake so that only 20% of the final amount of cellulase has appeared when 80% of the maximum amount of ammonia has been consumed.

Journal ArticleDOI
TL;DR: The relationship between the dimensions of a microbe and the accumulation of that microbe in porous, inorganic structures has been determined and those relationships were established by varying the physical parameters of the carriers as well as their chemical composition.
Abstract: The relationship between the dimensions of a microbe and the accumulation of that microbe in porous, inorganic structures has been determined. That relationship is dependent upon the cell dimensions, the mode of reproduction, and the pore diameter of the material. In order to achieve high accumulation of microbes that reproduce by fission, at least 70% of the pores of an inorganic carrier should have pore diameters in the range of one times the smallest major dimension through five times the largest major dimension of the cell. To achieve the highest accumulation of microbes that reproduce by budding, at least 70% of the pores should have pore diameters in the range of one times the smallest dimension of the cell and less than four times the largest cell dimension. These relationships were established by varying the physical parameters of the carriers as well as their chemical composition.

Journal ArticleDOI
TL;DR: Penicillium chrysogenum was immobilized in polyacrylamide gel prepared from 5% acRYlamide monomers (85% acrylamides and 15% N,N′‐methylene bisacryamide) and the activity of penicillin production of the washed mycelium decreased with repeated use, while the activity in the immobilized mycelia increased initially and decreased gradually with repeatedUse.
Abstract: Penicillium chrysogenum was immobilized in polyacrylamide gel prepared from 5% acrylamide monomers (85% acrylamide and 15% N,N'-methylene bisacrylamide). Penicillin produced from glucose by the immobilized mycelium was 17% of that produced by washed mycelium. However, the activity of penicillin production of the washed mycelium decreased with repeated use. On the other hand, the activity of the immobilized mycelium increased initially and decreased gradually with repeated use. The rate of oxygen uptake of the immobilized mycelium was about 30% of that of the washed mycelium. The immobilized mycelium required oxygen for the production of penicillin.

Journal ArticleDOI
TL;DR: Both the H‐ and L‐gels could convert 4‐AD to ADD in the presence of PMS, the L‐gel showing a slightly higher conversion rate, while the immobilized cells were more stable, the stability of H‐gel and L-gel being almost the same.
Abstract: A cell suspension in a water‐insoluble organic solvent (benzene: n‐heptane, 1 : 1 by volume) of Nocardia rhodocrous (previously induced to synthesize steroid Δ1dehydrogenase) rapidly catalyzed the stoichiometric oxidation of 4‐androstene‐3,17‐dione (4‐AD) to androst‐l,4‐diene‐3,17‐dione (ADD) in the presence of phenazine methosulfate (PMS). High levels of 4‐AD or PMS reduced the conversion rates. No appreciable decrease in the conversion rate was observed on adding aqueous buffer solution to the thawed ceils (up to 9.4 g water/g dry cell). The whole cells were immobilized by entrapment in a hydrophilic gel (H‐gel) or a lipophilic gel (L‐gel) by use of a water‐soluble or water‐insoluble photocrosslinkable prepolymer. The reticula of H‐ and L‐gel matrices were impregnated with water and organic solvent, respectively. Both the H‐ and L‐gels could convert 4‐AD to ADD in the presence of PMS, the L‐gel showing a slightly higher conversion rate. Various lines of evidence indicate that the limiting factor is the penetration rate of 4‐AD into gel particles for the H‐gel, and the penetration rate of PMS for the L‐gel. The catalytic activities decreased considerably after several successive runs with the free cell suspension system, while the immobilized cells were more stable, the stability of H‐gel and L‐gel being almost the same.

Journal ArticleDOI
TL;DR: An enzyme preparation that could detoxify parathion and eight other organophosphate pesticides was covalently bound to either porous glass or porous silica beads and indicated no loss in immobilized‐enzyme activity.
Abstract: An enzyme preparation that could detoxify parathion and eight other organophosphate pesticides was covalently bound to either porous glass or porous silica beads. This immobilized-enzyme system was examined for its use in detoxification of pesticides in production wastewaters. The kinetics of parathion hydrolysis were examined at flow rates up to 96 liter/hr and at influent substrate concentrations ranging from 10--250 mg/liter. The enzyme reactor was able to hydrolyze 95% or more of the parathion added to industrial wastewaters generated during its production, thus reducing the effluent parathion concentration to below 500 ppb. Laboratory continuous-flow experiments were conducted for 70 days with industrial wastewater and indicated no loss in immobilized-enzyme activity. The influence of pH, temperature, solvents, and detergents on enzyme stability and activity and enzyme reactor kinetics will be discussed.

Journal ArticleDOI
TL;DR: The location of the β‐glucosidase activity in a whole culture broth of the thermophilic organism Thermoactinomyces has been studied and there is evidence that it is an intracellular soluble enzyme(s).
Abstract: The location of the beta-glucosidase activity in a whole culture broth of the thermophilic organism Thermoactinomyces has been studied. Little beta-glucosidase activity was found in the culture filtrate, while the culture solids contained the major part of the activity of the whole culture broth. The activity does not appear to be adsorbed to the culture solids; rather there is evidence that it is an intracellular soluble enzyme(s). The pH and temperature optima for a crude beta-glucosidase preparation were determined to be pH 6.5 and 50-55 degrees C. Enzyme activity studies indicate that the same enzyme(s) accounts for the beta-glucosidase and the cellobiase activities. The validity of using the filter paper activity of culture filtrates from Thermoactinomyces to predict the total saccharification of cellulosic materials to glucose is discussed.


Journal ArticleDOI
TL;DR: In fed‐batch cultures, slow addition of cellulose at high concentrations can improve enzyme productivity by as much as 33% over a batch process.
Abstract: The scope in improving enzyme productivities from the cellulose fermentation process is examined in laboratory-scale fermentors. The maximum productivity (30 IU/liter hr) is attained in a continuous-culture process with cell recycle using modified medium containing 0.5% cellulose. Optimum dilution rate and recycle ratio are determined as 0.025 hr−1 and 1.2, respectively, for the process. The system is analyzed and steady-state equations for predicting enzyme protein concentrations in the fermentor are developed. In fed-batch cultures, slow addition of cellulose at high concentrations can improve enzyme productivity by as much as 33% over a batch process. The scope and results of using modified medium for cellulase production are also presented.

Journal ArticleDOI
TL;DR: By employing a two‐stage continuous‐culture system, some of the more important physiological parameters involved in cellulose biosynthesis have been evaluated with an ultimate objective of designing an optimally controlled cellulose process.
Abstract: By employing a two-stage continuous-culture system, some of the more important physiological parameters involved in cellulose biosynthesis have been evaluated with an ultimate objective of designing an optimally controlled cellulose process. The two-stage continuous-culture system was run for a period of 1350 hr with Trichoderma reesei strain MCG-77. The temperature and pH were controlled at 32°C and pH 4.5 for the first stage (growth) and 28°C and pH 3.5 for the second stage (enzyme production). Lactose was the only carbon source for the both stages. The ratio of specific uptake rate of carbon to that of nitrogen, Q(C)/Q(N), that supported good cell growth ranged from 11 to 15, and the ratio for maximum specific enzyme productivity ranged from 5 to 13. The maintenance coefficients determined for oxygen, MO, and for carbon source, MC, are 0.85 mmol O2/g biomass/hr and 0.14 mmol hexose/g biomass/hr, respectively. The yield constants determined are: YX/O = 32.3 g biomass/mol O2, YX/C = 1.1 g biomass/g C or YX/C = 0.44 g biomass/g hexose, YX/N = 12.5 g biomass/g nitrogen for the cell growth stage, and YX/N = 16.6 g biomass/g nitrogen for the enzyme production stage. Enzyme was produced only in the second stage. Volumetric and specific enzyme productivities obtained were 90 IU/liter/hr and 8 IU/g biomass/hr, respectively. The maximum specific enzyme productivity observed was 14.8 IU/g biomass/hr. The optimal dilution rate in the second stage that corresponded to the maximum enzyme productivity was 0.026 ∼ 0.028 hr−1, and the specific growth rate in the second stage that supported maximum specific enzyme productivity was equal to or slightly less than zero.

Journal ArticleDOI
TL;DR: A direct comparison of microcarrier cultures to roller bottle cultures showed equivalent interferon yields on a per cell basis and some apparent differences relating to optimum inducer concentrations and kinetics ofInterferon accumulation.
Abstract: A variety of diploid human fibroblast lines have been successfully grown to high densities (greater than 10(6) cell/ml) on recently developed microcarriers. Interferon induction using poly I.poly C and a superinduction procedure resulted in yields greater than 10,000 units/ml with one cell line. A direct comparison of microcarrier cultures to roller bottle cultures showed equivalent interferon yields on a per cell basis and some apparent differences relating to optimum inducer concentrations and kinetics of interferon accumulation.

Journal ArticleDOI
TL;DR: The growth and citric acid production kinetics of Saccharomycopsis lipolytica on glucose are investigated in an aerated stirred fermentor with a Michaelis–Menten‐type dependence on oxygen concentration.
Abstract: The growth and citric acid production kinetics of Saccharomycopsis lipolytica on glucose are investigated in an aerated stirred fermentor. Cellular growth first proceeds exponentially until exhaustion of ammonia in the fermentation medium. Cells then continue to grow at a reduced rate with a concomitant decrease in intracellular nitrogen content. Citric and isocitric acid production starts at the end of the growth phase. During about 80 hr excretion proceeds at a constant rate of 0.7 g/liter/hr for citric acid and 0.1 g/liter/hr for isocitric acid. The final citric and isocitric acid concentrations are 95 and 10 g/liter, respectively. During acid excretion cellular respiration accounts for 60 and 35% of consumed oxygen and glucose. Both acid and CO/sub 2/ production rates follow a Michaelis-Menten-type dependence on oxygen concentration with Michaelis-Menten constants of 0.9 and 0.15 mg/liter for acid and CO/sub 2/ productions, respectively.

Journal ArticleDOI
TL;DR: In this article, the authors evaluated one of the most promising of yeast that has flocculent characteristics and that is capable of fermentation at high sugar concentrations - Saccharomyces carlsbergensis (uvarum).
Abstract: The fermentation of alcohol from renewable substrates is attracting attention as a promising energy resource. Brazil has started to develop its fermentation industries for producing fuel ethanol from crops such as sugar-cane and cassava. However, before alcohol production costs become competitive, significant improvements in technology will have to be made. This paper evaluates one of the most promising of yeasts that has flocculent characteristics and that is capable of fermentation at high sugar concentrations - Saccharomyces carlsbergensis (uvarum). A maximum specific ethanol production rate of 0.58 g/g/hour was found in batch culture at high yeast concentrations and a value of 0.75 g/g/hour in continuous culture with recycle. It is suggested that where flocculation properties, as well as fermentation properties are required, the use of this yeast could well be justified.