scispace - formally typeset
Search or ask a question

Showing papers in "Biotechnology and Bioengineering in 1973"




Journal ArticleDOI
TL;DR: In this article, a mixture of soil and activated sludge organisms was used to degrade phenol which was inhibitory to microorganisms at higher concentrations, and the purpose of the experiments was to determine the kinetic parameters governing growth of the organisms by measuring growth rates in batch culture.
Abstract: A mixed culture derived from soil and activated sludge organisms was used to degrade phenol which was inhibitory to microorganisms at higher concentrations. The purpose of the experiments was to determine the kinetic parameters governing growth of the organisms by measuring growth rates in batch culture. To maintain a constant inoculum for the experiments inoculum was taken from a continuously operating continuous culture. Two populations were studied corresponding to two separate residence times in the continuous culture apparatus. One contained predominantly filamentous organisms, the other nonfilamentous. Five kinetic models were applied to the data and the best kinetic parameters for each model were determined by nonlinear least squares techniques. The models were then evaluated for best relative fit to the data. No significant differences were found between the models on the basis of fit and so a choice was made on the grounds of simplicity. A model proposed by Haldane was chosen as the best. No function however gave a satisfactory fit at the highest growth rates obtained. This experimental maximum in the plot of growth rate against substrate concentration was very sharp.

152 citations


Journal ArticleDOI
TL;DR: The oxygen uptake rate in mycelial pellets of Aspergillus niger was studied experimentally and theoretically and good agreement was found between the theoretical analysis and the experimental data.
Abstract: The oxygen uptake rate in mycelial pellets of Aspergillus niger was studied experimentally and theoretically. The specific rate of respiration of mycelial pellets was found to decrease significantly with increasing pellet size. The distribution of respiratory activity in the mycelial pellets was evaluated and the specific rate of respiration of disrupted mycelia showed adaptation to the concentration of oxygen in the medium. The decrease of the specific rate of respiration of the mycelial pellets could be estimated according to diameter, mycelial density, oxygen diffusivity, and adaptation to the concentration of oxygen. Good agreement was found between the theoretical analysis and the experimental data.

142 citations


Journal ArticleDOI
TL;DR: New methods of crosslinking enzyme molecules inside a matrix with or without an inactive protein are described, finding that fragile enzyme systems, for instance those using mobile cofactors, can be efficiently immobilized.
Abstract: New methods of crosslinking enzyme molecules inside a matrix with or without an inactive protein are described. Enzyme activity yields range between 30 and 80% of the activity of the untreated preparations. Even fragile enzyme systems, for instance those using mobile cofactors, can be efficiently immobilized. Increased resistance towards heat denaturation and proteolysis results.

124 citations


Journal ArticleDOI
TL;DR: In this article, a general procedure is presented for solving numerically for the substrate concentration profile and effectiveness factor utilizing the transformation method suggested by Na and Na. The use of a unique definition of the Thiele Modulus in these charts permits the clear illustration of the effects of substrate concentration and external mass transfer resistances on the overall effectiveness factor for the catalyst particle.
Abstract: The steady state, nonlinear diffusion equations which describe reactions in constrained enzyme solutions are of great interest in many biological and engineering applications. As in other types of nonlinear differential equations, exact analytical solutions do not exist except in some simplified cases. In this paper, a general procedure is presented for solving numerically for the substrate concentration profile and effectiveness factor utilizing the transformation method suggested by Na and Na. Design correlations for enzyme solutions constrained within spherical membranes are included. The use of a unique definition of the Thiele Modulus in these charts permits the clear illustration of the effects of substrate concentration and external mass transfer resistances on the overall effectiveness factor for the catalyst particle.

104 citations


Journal ArticleDOI
TL;DR: It appears that immobilized enzymes free of internal diffusional restrictions generally require the use of supports with pore sizes larger than those currently available in porous glass.
Abstract: Quantitative studies are reported of external film and internal pore diffusion effects for glucose oxidase immobilized on porous glass in a continuous, tubular, packed bed reactor. The McCune and Wilhelm model for external film diffusion is shown to describe these data over a particle Reynolds number range from 0.2 to 25. The Thiele analysis provides a good description of internal pore diffusion resistances. It appears that immobilized enzymes free of internal diffusional restrictions generally require the use of supports with pore sizes larger than those currently available in porous glass.

99 citations



Journal ArticleDOI
TL;DR: In this paper, a new correlation was given for the prediction of the volumetric coefficient for mass transfer (KLa) in stirred tanks from dispersed gas bubbles to basal salt solutions of ionic strengths representative of fermentation media.
Abstract: A new correlation is given for the prediction of the volumetric coefficient for mass transfer (KLa) in stirred tanks from dispersed gas bubbles to basal salt solutions of ionic strengths representative of fermentation media. The correlation includes the effects of both the operating parameters (agitation power per unit volume and gas superficial velocity) and the physicochemical properties of the system: interfacial tension, viscosity, density, diffusion, coefficient and, in particular, ionic strength. The effect of the latter was found to be most significant in the Newtonian systems of water‐like viscosity investigated; no previous correlations have included the effect of ionic strength. KLa values were determined by using a dissolved oxygen probe to monitor the steady‐state oxygen tension in continuous flow experiments, and/or the rate of change of oxygen tension in unsteady‐state semibatch experiments. In the latter cases, results were computed by a nonlinear, least squares computer program which fitted the experimental data to a model of probe transient response characteristics. The general applicability of the model and the computational procedure was verified by comparing the results to those obtained with the same electrolyte solution in the steady‐state mode. The experiments were run over a wide range of agitation power inputs, including those typical of both soluble‐ and insoluble‐substrate fermentations. The correlation appears to be valid for both oxygen mass transfer with and without homogeneous chemical reaction in the liquid phase; in the former case, for example, sulfite oxidation, knowledge of the chemical reaction enhancement factor is required. In addition to predicting oxygen transfer capabilities, the correlation may be used for other sparingly soluble gases of interest in fermentation systems, such as methane, hydrogen, and carbon dioxide.

95 citations


Journal ArticleDOI
TL;DR: A simplified model of cell metabolism, consisting of a series of linked reversible enzymatic reactions dependent on the concentration of a single external substrate, confirms the concept of a rate‐limiting step in biological systems as first proposed by Blackman.
Abstract: A simplified model of cell metabolism, consisting of a series of linked reversible enzymatic reactions dependent on the concentration of a single external substrate has been developed. The general mathematical solution for this system of reactions is presented. This general solution confirms the concept of a rate-limiting step, or “master reaction”, in biological systems as first proposed by Blackman. The maximum rate of such a process is determined by, and equal to, the maximum rate of the slowest forward reaction in the series. Of practical interest in modeling the growth rate of cells are three cases developed from the general model. The simplest special case results in the Monod equation when the maximum forward rate of one enzymatic reaction in the cell is much less than the maximum forward rate of any other enzymatic reactions. More realistic is the case where the maximum forward rates of more than one enzymatic reaction are slow. When two slow enzymatic reactions are separated from each other by any number of fast reactions that overall can be described by a large equilibrium constant, the Blackman form results: and A third case is that in which two slow enzymatic steps are separated by an equilibrium constant that is not large. Unlike the Monod and Blackman forms, which contain only two arbitrary constants, this model contains three arbitrary constants: The Monod and Blackman forms are special cases of this three constant form. In comparing equations with two arbitrary constants the Monod equation gave poorer fit of the data in most cases than the Blackman form. It is concluded that workers modeling the growth of microorganisms should give a t least as much consideration to the Blackman form as is given to the Monod equation.

94 citations


Journal ArticleDOI
TL;DR: No marked difference was observed between the native and immobilized aspartases with regard to optimum pH, temperature, concentration of Mn++, kinetic constants and heat stability.
Abstract: Various methods were tried for the immobilization of aspartase, and the preparation having the highest activity was obtained when partially purified aspartase from Escherichia coli was entrapped into polyacrylamide gel Iattice. Enzymatic properties of the immobilized aspartase were investigated and compared with those of the native aspartase. With regard to optimum pH, temperature, concentration of Mn++, kinetic constants and heat stability, no marked difference was observed between the native and immobilized aspartases. By employing an enzyme column packed with the immobilized aspartase, conditions for continuous production of L-aspartic acid from ammonium fumarate were investigated. When a solution of 1M ammonium fumarate (pH 8.5, containing 1mM MnCl2) was passed through the aspartase column at the flow rate of SV = 0.08 at 37°C, the highest rate of reaction was attained. From the column effluents, L-aspartic acid was obtained in a good yield.

Journal ArticleDOI
TL;DR: The integrated from of the rate equation has been shown to predict satisfactorily the progress of the reaction in a batch reactor using either soluble or immobilized penicillin amidase, and to predict the performance of a continuous feed stirred tank reactor containing immobilized enzyme.
Abstract: A rate equation has been derived to describe the hydrolysis of benzylpenicillin to 6-aminopenicillanic acid by penicillin amidase. The integrated from of the rate equation has been shown to predict satisfactorily the progress of the reaction in a batch reactor using either soluble or immobilized penicillin amidase. The rate equation was also used to predict the performance of a continuous feed stirred tank reactor containing immobilized enzyme. There was good agreement with experimental measurements.



Journal ArticleDOI
TL;DR: In this paper, a yeast lactase, Maxilact, was immobilized in crosslinked polyacrylamide using a bead-polymerization technique, and the polymer beads obtained, containing the entrapped enzyme, were used for the preparation of lactose-free milk.
Abstract: A yeast lactase, Maxilact, was immobilized in crosslinked polyacrylamide using a bead-polymerization technique. The polymer beads obtained, containing the entrapped enzyme, were used for the preparation of lactose-free milk. The binding yield of the enzyme and residual enzymic activity in the “enzyme beads” were studied as a function of the amounts of monomeric acrylamide and cysteine and bovine serum albumin present as protecting agents in the monomer-enzyme solution prior to polymerization. A maximum of about 75% of the enzyme could be immobilized using a 20% (w/v) solution of acrylamide plus N, N′-methylenebis-acrylamide, whereas the highest activity quotient (bound to free) of about 60% was observed on using a 25% solution. The presence of cysteine increased the activity by up to 30% and that of serum albumin up to about 15%.

Journal ArticleDOI
TL;DR: In this article, the authors showed that the organism takes up hydrocarbon accommodated in the aqueous phase as submicron droplets, and the cell/substrate yield for the linear growth phase, where growth was limited by the supply of the substrate, was much higher than the yield for exponential growth phase.
Abstract: Candida tropicalis was cultured with n-hexadecane, dispersed in water as submicron droplets, as the only carbon substrate; the emulsion being fed continuously into a fermentor containing only an aqueous medium (fed-batch culture). The results have demonstrated that the organism takes up hydrocarbon accommodated in the aqueous phase as submicron droplets. The cell/substrate yield for the linear growth phase, where growth was limited by the supply of the substrate, was much higher than the yield for the exponential growth phase.

Journal ArticleDOI
TL;DR: Xanthan gum, a heteropolysaccharide with unusual and useful properties, is now produced commercially by fermentation with Xanthomonas compestris NRRL B-1459 in a medium containing glucose, minerals, and a complex nitrogen source.
Abstract: Xanthan gum, a heteropolysaccharide with unusual and useful properties, is now produced commercially by fermentation with Xanthomonas compestris NRRL B–1459 in a medium containing glucose, minerals, and a complex nitrogen source—distillers' dried solubles (DDS). Understanding the kinetics of the fermentation should contribute to process improvements and increase the market potential for the gum. Earlier studies showed that although DDS determined initial growth rate, growth was stopped by some mechanism other than substrate exhaustion, probably an effect related to product formation. Product formation did not require active growth, but its rate increased with cell concentration. Specific product formation rate declined at high viscosities. Varying glucose concentration from 0.5 to 5.0% and dissolved O2 tension between 20 and 90% air saturated had no effect on the rates, but pH had to be maintained near 7 and temperature near 28°C to permit continued product formation. Xanthan yield could be explained by the energy required for growth and polymerization, that energy coming from dissimilation of the part of the carbohydrate substrate not converted to polymer.

Journal ArticleDOI
Bernard J. Abbott1, Allen Clamen1
TL;DR: Assessment of organic compounds assessed as potential substrates for single cell protein production and assignment of cost factors to the yield coefficients provided an estimation of the impact m and μ on biomass production costs.
Abstract: Various organic compounds were assessed as potential substrates for single cell protein production. Substrate evaluation was based on the costs associated with the substrate, oxygen, and heat yield coefficients: Ysub, Yo, and Ykcal, respectively. Yo, and Ykcal, were calculated from experimental values of Ysub, and from the elemental composition of bacterial cells. The dependence of the yield coefficients on the specific growth rate (μ) and maintenance coefficient (m) also was assessed. The analysis disclosed that m caused two- to threefold variations in the yield coefficients as μ was increased from 10% to 100% of μmax. The effect of different m values at constant specific growth rates also was determined. The value of m had a significant effect on the yield coefficients even at high specific growth rates. Assignment of cost factors to the yield coefficients provided an estimation of the impact m and μ on biomass production costs.

Journal ArticleDOI
TL;DR: A glutamic acid producing microorganism (Corynebacterium glutamicum) is entrapped in a polyacrylamide gel and these immobilized microorganisms were used to produce glutamic Acid in successive batches of fresh medium.
Abstract: A glutamic acid producing microorganism (Corynebacterium glutamicum) is entrapped in a polyacrylamide gel. These immobilized microorganisms were used to produce glutamic acid in successive batches of fresh medium. Free microorganisms similarly used produced much less glutamic acid under similar conditions.

Journal ArticleDOI
TL;DR: In this paper, the kinetic behavior of the immobilized invertase on these small pellets was investigated in a packed-bed reactor and an effect of interparticle diffusion on the Machaelis constant was observed and a correlation was proposed to evaluate the effect.
Abstract: Immobilized invertase was prepared by binding native invertase to a polyamine type ion-exchange resin. Kinetic behavior of the immobilized invertase on these small pellets was investigated in a packed-bed reactor. For low flow rates, an effect of interparticle diffusion on the Machaelis constant was observed and a correlation was proposed to evaluate the effect. At high flow rates, Michaelis and inhibition constants were determined and compared with those for native invertase. With large pellets, intraparticle diffusion was found to be important and at high substrate concentrations, the effectiveness factor exceeded unity. Good agreement was found between the theoretical analyses and the experimental data.


Journal ArticleDOI
TL;DR: Collagen was used as carrier for the immobilization of invertase, lysozyme, urease, glucose oxidase, penicillin amidase, Penicillin Amidase, and glucose isomerase.
Abstract: Collagen was used as carrier for the immobilization of invertase, lysozyme, urease, glucose oxidase, penicillin amidase, and glucose isomerase. Immobilization was accomplished by either impregnation of a preswollen collagen membrane with enzyme solution or electrocodeposition of collagen and enzyme from a collagen dispersion containing dissolved enzyme. The collagen-enzyme complexes prepared are in membrane form. Membranous collagen-enzyme complexes were used to construct biocatalytic reactors such as the capillaric coil modular reactor. Such biocatalytic reactors were used in a recirculation system for the conversion of substrates. The biocatalytic reactors showed initial decreases of activity to stable limits which are maintained over a large number of reactor volume replacements. The stable limits correspond to approximately 35% of the initial activities for lysozyme and invertase, 25% for urease, 15% for glucose oxidase. The mechanism of complex formation between collagen and enzyme involves multiple salt linkages, hydrogen bonds, and van der Waals interactions. This protein-protein interaction which leads to stable complexes by both impregnation and electrocodeposition processes is unique among the enzyme immobilization methods currently available.

Journal ArticleDOI
TL;DR: Carboxypolymethylene (Carbopol) was found to improve respiration rates in Aspergillus niger by as much as 200% as mentioned in this paper. But, the enhancement of other fermentation parameters, such as cellular growth and amylase production, was not observed.
Abstract: Of 24 different polymer and surfactant materials examined, a carboxypolymethylene (“Carbopol”) was found to cause enhancement of respiration rates in an Aspergillus niger culture by as much as 200%. Enhancement of other fermentation parameters, such as cellular growth and amylase production, was also observed. The enhancement effects of Carbopol were examined with clusters of spores and mold pellets. In the first case, it appears that the ionized carboxyl groups of Carbopol induced electrostatic repulsion among the spores thus initiating pulp growth with increased interfacial area of contact between the mold and the nutrient medium. In the second case, the Carbopol additive formed a thin film attached to the surface of the pellets which seemed to be responsible for an increased rate of potassium transport and, hence, fermentation yields. Additive utilization as substrate and physiological changes in the culture were not observed in these cases. It was also found that the probability of pellet formation, the size of pellets formed, and the number of spores per pellet can be correlated to the energy input to the fermentation system.

Journal ArticleDOI
TL;DR: The predation of bacteria by protozoan in both continuous and batch cultures was studied using experimental and modeling techniques and the models studied were partially successful in simulating the empirical data.
Abstract: The predation of bacteria by protozoan in both continuous and batch cultures was studied using experimental and modeling techniques The predator organism was the ciliate, Tetrahymena pyriformis The bacterium, Aerobacter aerogenes, served as the prey Several batch growth responses were observed each initiated at a different nutrient level Continuous cultures were conducted over a range of dilution rates The models studied were partially successful in simulating the empirical data Deviations between the model and the actual population behavior are discussed and possible explanations for the differences proposed

Journal ArticleDOI
TL;DR: Methods for the production of protein concentrates, with a low content of nucleic acid, in kilogram quantities from yeast have been studied with the aid of equipment designed for operation on pilot‐plant scale and high extractability of protein and nucleic Acid was only obtained after mechanical disintegration.
Abstract: Methods for the production of protein concentrates, with a low content of nucleic acid, in kilogram quantities from yeast have been studied with the aid of equipment designed for operation on pilot-plant scale. The influence of drum drying and mechanical disintegration on the nutritive value of the yeast was also investigated. Drum drying and mechanical disintegration improved the nutritive value of the yeast but high extractability of protein and nucleic acid was only obtained after mechanical disintegration. Protein concentrates without and with cell walls were produced from mechanically disintegrated yeast. The different fractions which were obtained when separating cell walls and precipitating protein by heating at alkaline pH, were analyzed. After protein precipitation, about 90% of the RNA could be precipitated from the supernatant by addition of acid, giving a product containing 50% RNA of the dry weight. The protein precipitate obtained after cell wall separation had an RNA content of less than 2% and contained 70–l75% of the amino acids in the starting yeast material. Protein concentrates containing cell walls were produced by precipitating protein by heating at alkaline pH directly after mechanical disintegration. The content of RNA was about 2% and the yield of amino acids was 70–80%. It was found that the nutritive value of the protein concentrate was higher than that of the starting yeast material. To produce such a protein concentrate on a large scale, the process described can probably be employed.

Journal ArticleDOI
TL;DR: The thermal stability of glucose oxidase in solution was studied as a function of time and temperature between 37–60°C and both polymer concentration and compositions were shown to be critical parameters.
Abstract: The thermal stability of glucose oxidase in solution was studied as a function of time and temperature between 37–60°C. As expected, the rate of thermal inactivation increased with temperature and at 60°C more than 80% of the enzyme's activity was lost after 0.5 hr incubation. Similar stability measurements on enzyme solutions containing water soluble synthetic polymers showed that several of the polymers significantly enhanced the thermal stability of glucose oxidase. Copolymers of vinyl acetate with either vinyl pyrrolidone or vinyl alcohol were found to be particularly effective. The molecular weight of the added polymers was found to be unimportant in the stabilization process but both polymer concentration and compositions were shown to be critical parameters.

Journal ArticleDOI
TL;DR: In this paper, the authors investigated the kinetics of C. tropicalis growth with pure n-hexadecane as a dispersed phase substrate and found two distinct growth phases: in the first phase, exponential growth was independent of stirrer speed.
Abstract: The kinetics of C. tropicalis growth were investigated with pure n-hexadecane as dispersed phase substrate. Two distinct growth phases were found: In the first phase, exponential growth was independent of stirrer speed. The onset of the second phase, one of linear growth, was determined by stirrer speed. By the use of two different fermenter types, it was shown that the drop size of the dispersed phase was not primarily responsible for the observed kinetics. It was considered that the formation of biological flocs determined the observed growth pattern. This was substantiated by the results of continuous cultures in the different fermenter types, with various substrate concentrations.

Journal ArticleDOI
TL;DR: In this article, a study was made to determine the controlling mass transfer resistance in the overall reaction rate for conversion of maltose to glucose, catalyzed by glucoamylase immobilized onto porous glass.
Abstract: A study was made to determine the controlling mass transfer resistance in the overall reaction rate for conversion of maltose to glucose, catalyzed by glucoamylase immobilized onto porous glass. For normal operation of a packed column and air-stirred batch reactor, the rate controlling step was found to be the internal resistance of simultaneous pore diffusion and chemical reaction. Experimental effectiveness factors were determined and are compared with those derived from a theoretical diffusion model based on Michaelis-Menten kinetics. Also given are temperature and pH relationships for the free and immobilized glucoamylase.

Journal ArticleDOI
TL;DR: A simple two‐parameter model is developed describing the degree of repression or induction in fermentation media.
Abstract: The synthesis of extracellular enzymes by microorganisms frequently occurs under genetic control. A simple two-parameter model is developed describing the degree of repression or induction in fermentation media. The case of substrate utilization by an extracellular enzyme was analyzed for a vegetable oil-lipase-yeast system. It is shown that fatty acids released by the lipase may accumulate in the early stage of growth and exert an influence on the limiting after which relatively little repression or induction takes place. Expressions are also derived for growth and extracellular enzyme synthesis in single-and multistage continuous cultures. When the cells grow on a directly available soluble substrate, the specific enzyme synthesis is maximal at low dilution rates in the case of repression and at high dilution rates in the case of induction. If the substrate is not directly available, a single continuous stirred tank reactor stage may not be sufficient for efficient substrate utilization; for fermentation processes where an insoluble has to be broken down before the cells can assimilate it, a plug flow type fermentor rather than a mixed chemostat may prove more satisfactory.

Journal ArticleDOI
TL;DR: A periodic countercurrent whole beer process was developed for the recovery of novobiocin to eliminate the high cost of mycelium filtration and accompanying antibiotic losses in the filter cake.
Abstract: A periodic countercurrent whole beer process was developed for the recovery of novobiocin to eliminate the high cost of mycelium filtration and accompanying antibiotic losses in the filter cake. In such a process screened, but unfiltered, novobiocin fermentation beer is contacted with a special grade of an anionic-exchange resin in a series of specially designed, well mixed columns. Each column is fitted with a screen sized to retain resin within the column but allow the cells to press. Periodically the lead column is isolated from the column train, washed free of beer solids, and eluted. A freshly eluted column is placed in the trail position to allow countercurrent operation. The eluate is then processed to crystalline product. A mathematical model for the sorption of novobiocin was developed based on a suitable continuity equation and mass transfer and equilibrium relationships determined in the laboratory. Digital computations of this model correlated well with laboratory and pilot plant data, and predicted well the performance of the production units. This simulation has been in continued use to predict and reoptimize plant operation as process changes (such as improved beer titers, increased production rate, and the evaluation of superior ion-exchange resins) occurred or were anticipated.