Showing papers in "Biotechnology and Bioengineering in 1982"

The mechanism of uranium biosorption by Rhizopus arrhizus.

Abstract: Biosorption of elements is a little understood phenomenon exhibited by some types of even nonliving microbial biomass. A common fungus Rhizopus arrhizus has been reported to take up uranium from aqueous solutions to the extent of 180 mg U(6+)/g. The mechanism of uranium sequestering by this type of biomass was studied by using experimental techniques such as electron microscopy, x-ray energy dispersion analysis, IR spectroscopy, and supporting evidence was obtained for a biosorption mechanism consisting of at least three processes. Uranium coordination and adsorption in the cell-wall chitin structure occur simultaneously and rapidly whereas precipitation of uranylhydroxide within the chitin microcrystalline cell-wall structure takes place at a lower rate. Interference of Fe(2+) and Zn(2+) coions with uranium biosorption is indicated.

Conversion of d-xylose to ethanol by the yeast pachysolen tannophilus

Abstract: A method has been discovered for converting D-xylose to ethanol relying on the unique ability of the yeast Pachysolen tannophilus to ferment this five-carbon sugar without the use of added enzymes. This process will be particularly useful in the production of ethanolic fuel from plant biomass.

Hydrocracking of the oils of Botryococcus braunii to transport fuels.

Abstract: Hydrocarbon oils of the alga Botryococcus braunii, extracted from a natural “bloom” of the plant, have been hydrocracked to produce a distillate comprising 67% gasoline fraction, 15% aviation turbine fuel fraction, 15% diesel fuel fraction, and 3% residual oil. The distillate was examined by a number of standard petroleum industry test methods. This preliminary investigation indicates that the oils of B. braunii are suitable as a feedstock material for hydrocracking to transport fuels.

Kinetic studies of enzymatic hydrolysis of insoluble cellulose: analysis of the initial rates.

Abstract: A study was conducted on the kinetics of enzymatic hydrolysis of pure insoluble cellulose using unpurified culture filtrate Trichoderma reesei, with the emphasis on the initial reaction period. The initial hydrolysis rate and extent of enzyme (soluble protein)adsorption, either apparent or initial, were evaluated under various experimental conditions. It has been found that the various mass-transfer steps do not control the overall hydrolysis rate and that the hydrolysis rate is mainly controlled by the surface reaction step promoted by the adsorbed enzyme. It has also been found that the initial hydrolysis rate strongly depends on the initial extent of soluble protein adsorption and the effectiveness of the adsorbed soluble protein to promote the hydrolysis. The initial extent of soluble protein adsorption, in turn, is related to the initial cellulose concentration, enzyme concentration, and specific surface area of cellulose, whereas the effectiveness of the initially adsorbed soluble protein to promote the derived to interrelate these parameters without resorting to the Michaelis-Menten kinetics. The present result appear to imply that the role of enzyme-substrate complex formation should not be ignored in deriving a mechanistic kinetic model for enzymatic hydrolysis of cellulose.

Ethanol production by extractive fermentation

Abstract: The ideal method to produce a terminal metabolite inhibitor of cell growth and production is to remove and recover it from the fermenting broth as it formed. Extractive fermentation is achieved in the case of ethanol production by coupling both fermentation and liquid-liquid extraction, The solvent of extraction is 1-dodecanol (or a mixture 1-dedecanol, 1-tetradecanol); study of the inhibitory effect of primary aliphatic alcohols of different chain lengths shows that no growth is observed in the presence of alcohols which have between 2 and 12 carbons. This effect is suppressed when the carbon number is 12 or higher. A new reactor has been used-1 pulsed packed column. Pulsation is performed pneumatically. Porous material used as a package adsorbs the cells. The fermentation broth is pulsed in order to (1) increase the interfacial area between the aqueous phase and the dodecanol, (2) decrease gas holdup. Alcoholic fermentation, performed at 35 degrees C on glucose syrup, permits the total utilization of glucose solution of 409 g/L with a yeast which cannot-in classical process- completely use solutions with 200 g/L of glucose. The feasibility of a new method of fermentation coupling both liquid-liquid extraction and fermentation is demonstrated. Extension of this method is possible to any microbial production inhibited by its metabolite excretion.

Enzymatic hydrolysis of cellulose and various pretreated wood fractions

Abstract: Three strains of Trichoderma-T. reesei C30, T. reesei QM9414, and Trichoderma species E-58-were used to study the enzymatic hydrolysis of pretreated wood substrates. ach of the culture filtrates was incubated with a variety of commercially prepared cellulose substrates and pretreated wood substrates. Solka floc was the most easily degraded commercial cellulose. The enzyme accessibility of steam-exploded samples which had been alkali extracted and then stored wet decreased with the duration of the steam treatment. Air drying reduced the extent of hydrolysis of all the samples but had a greater effect on the samples which had previously shown the greatest hydrolysis. Mild pulping using 2% chlorite increased the enzymatic hydrolysis of all the samples. Steam explosion was shown to be an excellent pretreatment. The results indicate that the distribution of the lignin as well as the surface area of the cellulosic substrate are important features in enzymatic hydrolysis.

Adsorption of cellulase on cellulose: Effect of physicochemical properties of cellulose on adsorption and rate of hydrolysis.

Abstract: In the cellulase-cellulose reaction system, the adsorption of cellulase on the solid cellulose substrate was found to be one of the important parameters that govern the enzymatic hydrolysis rate of cellulose. The adsorption of cellulase usually parallels the rate of hydrolysis of cellulose. The affinity for cellulase varies depending on the structural properties of cellulose. Adsorption parameters such as the half-saturation constant, the maximum adsorption constant, and the distribution coefficient for both the cellulase and cellulsoe have been experimentally determined for several substrates. These adsorption parameters vary with the source of cellulose and the pretreatment methods and are correlated with the crystallinity and the specific surface area of cellulose substrates. The changing pattern of adsorption profile of cellulase during the hydrolysis reaction has also been elucidated. For practical utilization of cellulosic materials, the cellulose structural properties and their effects on cellulase adsorption, and the rate of hydrolysis must be taken into consideration.

Processes governing primary biofilm formation.

Abstract: Biofilm accumulation under turbulent flow condition on the surface of a circular tube is the net result of several process including the following: (1) transport and firm adhesion of soluble components and microbial cell to the surface; (2) metabolic conversions within the biofilm in cluding growth and maintenance decay process; (3) detachment of portions of the biofilm and reentrainment in the bulk fluid. Experiments in tabular reactor were designed to measure the rates of these process during the early stages of biofilm accumulation as a function of the Reynolds number and suspended biomass concentration. Results indicate deposition (i.e., combined transport and adsorption) is only important in the very early stages of biofilm accumulation and is significantly influenced by negligible for the thin biofilms encountered in these experiments. Net biofilm production rates in all experiments decrease to same level and this level is not affected by changes in Reynolds number or suspended biomass concentration. Biofilm detachment rate increases continuously with biofilm accumulation and with increasing Reynolds number.

Accumulation of heavy metals in unicellular algae

Abstract: A model for metal uptake by microorganisms based on surface adsorption has been developed, and then applied to the uptake of cadmium by Chlorella vulgaris. A linear equilibrium relationship between metal in the solution and that adsorbed on the cell surface is assumed and confirmed at low cadmium concentrations by short-term uptake experiments. When it incorporates a description of cell growth, the model predicts an initial rapid uptake and a subsequent slow uptake. Such behavior has often been observed in experiments with growing microorganisms. This indicates that the slow uptake, sometimes thought to be active or metabolic, could be due to the simultaneous effects of growth and surface adsorption. The model shows that initial metal uptake is fast and approaches equilibrium within a few seconds. This prediction is in agreement with experimental results in a batch system: Equilibrium is reached before the first samples are taken (at 10 min) and there is then no measurable change until growth provides a significant increase in cell surface (after several hours). Thus the equilibrium constant can be calculated from experimental results of uptake at 10 min. The equilibrium is found to be affected by phosphate concentration; the amount of cadmium adsorbed on the cell decreases as the concentration of phosphate is increased. Long-term uptake experiments in growing cultures show a greater metal accumulation than predicted by the adsorption model, suggesting the involvement in the slow long-term uptake of some mechanism other than adsorption. This is confirmed by experiments in which uptake in cultures exposed to cadmium throughout the growth period is compared with short-term uptake in similar cultures grown in the absence of cadmium. The modeling approach to metal adsorption provides a basis for further development. A model combining description of adsorption and of intracellular accumulation is necessary to provide a more complete description. Such a model, with precise definitions of system parameters and means of evaluating these parameters from experimental results, will be a powerful tool in investigation of metal uptake by microorganisms.

The mechanism of thorium biosorption by Rhizopus arrhizus.

Abstract: Inactive cells of Rhizopus arrhizus have been documented to exhibit a high thorium biosorptive uptake (170 mg/g) from aqueous solutions. The mechanism of thorium sequestering by this biomass type was investigated following the same method as for the uranium biosorption mechanism. The thorium sequestering mechanism appeared somewhat different from that of uranium. Experimental evidence is presented which indicates that, at optimum biosorption pH (4), thorium coordinates with the nitrogen of the chitin cell wall network and, in addition, more thorium is absorbed by the external section of the fungal cell wall. At pH 2 the overall thorium uptake is reduced. The kinetic study of thorium biosorption revealed a very rapid rate of uptake. Unlike uranium at optimum solution pH, Fe(2+) and Zn(2+) did not interfere significantly with the thorium biosorptive uptake capacity of R. arrhizus.

Effects of ethanol and other alkanols on the glucose transport system of Saccharomyces cerevisiae.

Abstract: Maltose transport in S. cerevisiae was inhibited by ethanol and other alkanols in a non-competitive way. The Michaelis constant, Km, for the sugar, with or without alkanols was 5.9 mM, whereas the maximum trans port capacity, Vmax, decreased exponentially with alkanols concentration. The inhibitory capacity was positively correlated with the lipid solubility of the alkanols, indicating that inhibition is due to an alteration of the lipid environment of the maltose transport system in the plasma membrane.

Comparative performance of biofilm reactor types.

Abstract: Development of a unified model of biofilm-reactor kinetics is based on substrate-utilization kinetics, mass transport, biofilm growth, and reactor analysis. The model is applied to steady-state conditions for complete-mix, fixed-bed, and fluidized-bed reactors with and without recycle. The results of modeling experiments demonstrate that simple loading factors and kinetic relationships are insufficient to describe the performance of a variety of biofilm processes. Instead, the interactions among utilization kinetics, biofilm growth, and reactor configuration determine the performance. For example, fluidized-bed reactors can achieve superior performance to complete-mix and fixed-bed reactors because the biofilm is evenly distributed throughout the reactor while the liquid regime has plug-flow characteristics. When it is possible, experimental results which demonstrate key concepts are presented.

Properties of hydrocarbon-in-water emulsions stabilized by Acinetobacter RAG-1 emulsan.

Abstract: Emulsan is a polymeric extracellular emulsifying agent produced by Acinetobacter RAG-1. Hydrocarbon-in-water emulsions (V(f) of hydrocarbon of 0.01-0.10) were stabilized by small quantities of emulsan (0.02-0.2 mg/mL). Although both aliphatic and aromatic hydrocarbon emulsions were stabilized by emulsan, mixtures containing both aliphatics and aromatics were better substrates for emulsan than the individual hydrocarbon by themselves. The emulsan remained tightly bound to the hydrocarbon even after centrifugation as determined by (a) residual emulsan in the aqueous phase and (b) the fact that the resulting "cream" readily dispersed in water to reform stable emulsions. With hexadecane-to-emulsan weight ratio of 39 and 155, the noncoalescing oil droplets had average droplet diameters of 2.0 and 4.0 microm, respectively. Dialysis studies showed that the water-soluble dye Rhodamine B adsorbed tightly to the interface of hexadecane-emulsan droplets although the dye did not bind to either hexadecane or emulsan alone. At saturating concentrations of dye, 2.2 micromol of dye were bound per mg emulsan.

Effect of compression milling on cellulose structure and on enzymatic hydrolysis kinetics

Abstract: The changes in the cellulose structure by compression milling were studied and expressed in terms of crystallinity, accessibility, specific surface area, and degree of polymerization. The kinetic parameters, maximum reaction rate, and Michaelis constant were determined experimentally. Based on the experimental results a two-phase model, which is based on the degradation of cellulose by simultaneous actions of the cellulase complex on the crystalline and amorphous phases, is proposed. The relationships between cellulose accessibility and the kinetic parameters were compared with those predicted by the model. A good agreement was found, although the two-phase hypothesis is a simplification of the true state of order in cellulose.

Biogas production from anaerobic digestion of Spirulina maxima algal biomass

Abstract: The photosynthetic spectrum of solar energy could be exploited for the production of chemical energy of methane through the combined algal-bacterial process. In this process, the algae are mass produced from light and from carbon in the first step. The algal biomass is then used as a nutrient for feeding the anaerobic digester, in the second step, for the production of methane by anaerobic bacteria. The carbon source for the production of algal biomass could be either organic carbon from wastewaters (for eucaryotic algae), or carbon dioxide from the atmosphere or from the combustion exhaust gases (for both prokaryotic and eukaryotic algae). The technical feasibility data on the anaerobic digestion of algal biomass have been reported for many species of algae including macroscopic algae and microscopic algae. Research being conducted in the authors' laboratory consists of using the semimicroscopic blue-green alga Spirulina maxima as the sole substrate for this combined algal-bacterial process. This species of alga is very attractive for the process because of its capability of using the atmospheric carbon dioxide as carbon source and its simple harvesting methods. Furthermore, it appeared that the fermentability of S. maxima is significantly higher than other microscopic algae. This communication presents themore » results on the anaerobic inoculum development by the adaptation technique. This inoculum was then used for the semicontinuous anaerobic digestion of S. maxima algal biomass. The evolutions of biogas production and composition, biogas yield, total volatile fatty acids, alkalinity, ammonia nitrogen, pH, and electrode potential were followed.« less

Studies on immobilized Saccharomyces cerevisiae. I. Analysis of continuous rapid ethanol fermentation in immobilized cell reactor.

Abstract: Rapid fermentation of cane molasses into ethanol has been studied in batch, continuous (free-cell cell-immobilized systems) by a strain of Saccharomyces cerevisiae at temperature 30/sup 0/C and pH 5.0. The maximum productivity of ethanol obtained in immobilized system was 28.6 g L/sup -1/ h/sup -1/. The cells were immobilized by natural mode on a carrier of natural origin and retention of 0.132 g cells/g carrier was achieved. The immobilized-cell column was operated continuously at steady state over a period of 35 days. Based on the parameter data monitored from the system, mathematical analysis has been made and rate equations proposed, and the values of specific productivity of ehtanol and specific growth rate for immobilized cells computed. It has been established that immobilized cells exhibit higher specific rate of ethanol formation compared to free cells but the specific growth rate appears to be comparatively low. The yield of ethanol in the immobilized-cell system is also higher than in the free-cell system.

Effect of surface shear stress on the attachment of Pseudomonas fluorescens to stainless steel under defined flow conditions.

Abstract: The ease with which microorganisms grow on submerged surfaces is the cause of certain industrial problems (i.e. fouling and corrosion of pipelines, heat exchangers, cooling towers, etc.). Application of the radical-flow growth chamber to the study of the initial stages of bacterial adhesion to surfaces under flowing conditions is reported. The adhesive properties of the bacterium Pseudomonas fluorescens (NCIB9046) to stainless steel (type AISI 316 were found to highly dependent on surface shear stress and the time and concentration of cells used in the incubation procedure. Maximum levels of adhesion occurred in zones of lowest surface shear stress, particularly less than 6-8 Nm/sup -2/, Adhesion was still noticeable to shear stresses even up to 130 Nm/sup -2/. Significant detachment of cells from a monolayer attached under static conditions was found to occur at surface shear stresses in excess of 10-12 Nm/sup -2/. (JMT)

Oxygen mass transfer into aerated CMC solutions in a bubble column.

Abstract: Differing findings on the volumetric mass transfer coefficients k(L)a in CMC solutions in bubble column bioreactors have been reported in the literature. Therefore, oxygen mass transfer was studied again in CMC solutions in a 14-cm-i.d. x 270-cm-height bubble column using different spargers. The k(L)a values were determined along with the dispersion coefficients by fitting the prediction of the axial dispersed plug model with the experimental oxygen concentration profiles in the liquid phase. Surprisingly, the obtained liquid phase dispersion coefficients for CMC solution are higher than one would expect from correlations. The k(L)a data depend largely on the flow regime. In general, they are lower than those reported in the literature. The data for developing slug and established slug flow are dependent on the gas velocity and the effective viscosity of the solution and can br correlated by a simple correlation. This correlation describes k(L)a values measured on fermentation broth of Penicillium chrysogenum with striking agreement.

Evaluation of crude dextran as phase-forming polymer for the extraction of enzymes in aqueous two-phase systems in large scale.

Abstract: A detailed study of the influence of crude dextran on enzyme extractions in aqueous phase systems is presented in this article. The physical parameters of crude dextran, a purified T-500 fraction from Pharmacia, and a hydrolyzed crude dextran are compared and their influence on the phase system parameters investigated. Initially there is a drastic increase in the viscosity of the lower dextran-rich phase and a significant shift in the macroscopic structure of these phases, observed as the "gel-forming" properties of the dextran phases. The latter can be important for the partition of any enzyme by influencing the effect of phosphate concentration on the partition of proteins, although these experiments show that the partition coefficient of several enzymes is not much altered. The partition parameters allow the substitution of Dextran T-500 fractions by crude dextran or unfractionated, slightly hydrolyzed fractions. Using crude dextrans the performance and technical realization of enzyme extraction processes are demonstrated for pullulanase from Klebsiella pneumoniae and formate dehydrogenase from Candida boidinii.Both enzymes were recovered in comparable high yields. The equipment performance was quite good, as indicated by the high throughput values of the separators employed. Especially when using nozzle separators for phase separation there is a better performance in comparison to the Dextran T-500 fraction. No serious technical problems were encountered when replacing the expensive fractionated dextran with a crude dextran. In this way aqueous two-phase systems containing dextran become more feasible for enzyme purification from an economic point of view. The price of about 1.30 German marks (DM) per liter for a useful phase system already appears acceptable for the production of valuable intracellular enzymes.

Exploitation of plant cells for the production of natural compounds.

Abstract: A four-step strategy is presented which allows the establishment of plant cell cultures producing high yields of secondary plant products. The application of suitable methods (radioimmunoassay, fluorescence screening) for the selection of overproducing differentiated plants and cell colonies is stressed. By precursor feeding and hormone application, plant cell cultures can greatly be influenced in their production behavior. A highly sensitive, selective regulatory mechanism for the uptake and storage of alkaloids in Catharanthus vacuoles is demonstrated. Overproducing variant cell strains are so far the most promising tool for the future biotechnological application of the plant cell culture method.

Analysis and optimization of methods using water-soluble carbodiimide for immobilization of biochemicals to porous glass.

Abstract: Two methods employing a water-soluble carbodiimide for carboxyl activation were investigated for the immobilization of biochemicals to succinamidopropyl-porous glass beads. Immobilization using the simultaneous method (simultaneous addition of carbodiimide and nucleophilic ligand to the beads) and large excess of carbodiimide and a small nucleophile should result in covalent binding to all accessible carboxyl groups. Results obtained with glycine methyl ester indicated that 40% of the total surface carboxyl groups were sterically accessible. Using these reaction conditions with the protein, chymotrypsinogen, suggests that a surface monolayer is immobilized. although far fewer sites are required assuming single point attachment. For ligands containing carboxyl groups and several nucleophilic groups (e. g., enzymes), however, biological inactivation may occur using the simultaneous method. Consequently, a sequential method (activation of the surface with carbodiimide followed by washing and addition of the biochemical to be immobilized) was optimized. Using optimal conditions (20 min activation time at pH 4.75 and room temperature; 2 min wash at pH 7 and 0 degrees C) and 0.1M carbodiimide, nearly half of the accessible surface sites remained in the O-acylisourea form and reacted with glycine methyl ester upon its addition. The amount of surface loading as a function of activation time was consistent with a model constructed using rate constants for O-acylisourea formation and hydrolysis previously derived from solution studies with acetic acid [Swaisgood and Natake, J. Biochem 74, 77 (1973)]. Measurement of reaction rates with glycine methyl ester following surface activation suggests that the rate of reaction with amino groups is at least eightfold greater than the hydrolysis rate. Either immobilization procedure gave comparable enzyme loading and specific activities for the case of sulfhydryl oxidase.

Methane from cattle waste: Effects of temperature, hydraulic retention time, and influent substrate concentration on kinetic parameter (k)

Abstract: The effects of temperature (35 and 55 degrees C), influent volatile solids (VS) concentration (S(0) = 43, 64, 82, 100, and 128 kg VS/m(3)) and hydraulic retention time (HRT = 4, 5, 8, 10, 15, and 25 days) on methane (CH(4)) production from cattle waste were evaluated using 3-dm(3) laboratoryscale fermentors. The highest CH(4) production rate achieved was 6.11 m(3) CH(4) m(-3) fermentor day(-1) at 55 degrees C, four days HRT, and S(0) = 100 kg VS/m(3). Batch fermentations showed an ultimate CH(4) yield (B(0)) of 0.42 m(3) CH(4)/kg VS fed. The maximum loading rates for unstressed fermentation were 7 kg VS m(-3) day(-1) at 35 degrees C and 20 kg VS m(-3) day(-1) at 55 degrees C. The kinetic parameter (K, an increasing K indicates inhibition of fermentation) increased exponentially as S(0) increased, and was described by: K = 0.8 + 0.0016 e(0.06S(0) ). Temperature had no significant effect on K for S(0) between 40 and 100 kg VS/m(3). The above equation predicted published K values for cattle waste within a mean standard error of 7%.

Production of microbial lipid: Effects of growth rate and oxygen on lipid synthesis and fatty acid composition of Rhodotorula gracilis.

Abstract: Microbial lipids produced by Rhodotorula gracilis NRRL Y-1091 grown in continuous culture under nitrogen-limiting condition were evaluated and the effects of growth rate and oxygen concentration on the degree of unsaturatoin of fatty acids studied. As the growth rate increased the protein content of the biomass increased but cell biomass, lipid content, and lipid productivity decreased; the specific lipid production rate remained constant at about 0.012 g lipid/g dry biomass/h. The maximum lipid content recorded was 49.8% (w/w) of the cell mass at a growth rate of 0.02 h(-1). The growth rate also affected fatty acid composition; polyunsaturated fatty acids (C18:2 and C18:3) increaded with growth rate while other fatty acids (C16:0, C18:0, C18:1) decreased. Increase in oxygen concentration between 5 and 234muM increased the lipid content without significantly affecting its degree of unsaturation. On the other hand, the degree of unsaturation was significantly affected by specific oxygen uptake rate for this obligate aerobe, Rh. gracilis.

Strategies for penicillin fermentation in tower–loop reactors

Abstract: Since it has not been possible to produce penicillin in tower-loop reactors with highly viscous filamentous molds of Penicillium chrysogenum which are employed in stirred-tank reactors, a new strategy has been developed to avoid the formation of this morphology and to use the pellet form of the fungi. When employing definite impeller speeds in the subculture in connection with definite inoculum amounts and substrate concentrations in the main culture (bubble column), it is possible to generate a suspension of isolated small pellets, which shows a low broth viscosity up to a sediment content of 45% over the entire fermentation time. Volumetric mass-transfer coefficients k(L)as are by a factor of 4 to 5 higher in these pellet suspensions than in filamentous broths. It was easy to maintain the necessary oxygen supply for penicillin production in these pellet suspensions. Under these conditions the specific penicillin productivities were higher with regard to power input (up to 90%), biomass, and consumed substrate than in the stirred-tank reactors with highly viscous filamentous morphology of the fungi. Under nonoptimized operating conditions the absolute penicillin production in the tower loop was 35% lower than in the stirred-tank reactor due to lower possible biomass concentrations. The separation of the biomass, and therefore the penicillin recovery, is much simpler when employing pellets. It is shown how the particular mass transfer resistances at the gas/liquid and liquid/pellet interfaces and within the pellets change with the pellet diameter. There should be a particular pellet diameter at which penicillin productivity has its maximum. These investigations indicate that the use of tower-loop reactors can, in the future, be an alternative for more economical penicillin production methods.

Interactions between amino-acid-degrading bacteria and methanogenic bacteria in anaerobic digestion

Abstract: The degradation of amino acids in anaerobic digestion was examined in terms of the interactions between amino-acid-degrading bacteria and methanogenic bacteria. Certain amino acids were degraded oxidatively by dehydrogenation, with methanogenic bacteria acting as H(2) acceptors. The inhibition of methanogenesis by chloroform also inhibited the degradation of these amino acids and/or caused variations in the composition of volatile acids produced from them. The presence of glycine reduced the inhibitory effect caused by chloroform, probably because glycine acted as an H(2) acceptor in place of methanogenic bacteria. This fact suggested that the coupled oxidation-reduction reactions between two amino acids-one acting as the H(2) donor and the other acting as the H(2) acceptor-may occur in the anaerobic digestion of proteins or amino-acid mixtures. The conversion of some proteins to volatile acids was not affected when methanogenesis was inhibited by chloroform. This suggested that the component amino acids of proteins may be degraded by the coupled oxidation-reduction reactions and that the degradation of proteins may not be dependent on the activity of methanogenic bacteria as H(2) acceptors.

Predominant role of hydrocarbon solubilization in the microbial uptake of hydrocarbons.

Abstract: Using EDTA and proteolytic enzymes to suppress hydrocarbon solubilization, direct evidence is presented in support of the mechanism of liquid hydrocarbon uptake by microbial cells predominantly from the solubilized or accommodated substrate. EDTA (2-5mM) strongly inhibited growth of three yeast species and one bacterial species on n-hexadecane and the inhibition was removed by surfactant-emulsified and surfactant-solubilized alkane and also by excess addition of Ca(2+). EDTA had no inhibitory effect on the growth of the organisms on soluble substrates such as sodium acetate and nutrient broth or on n-pentane, a volatile alkane which was primarily transported by diffusion from gas phase. EDTA was shown to have no significant effect on the adsorption of cells on alkane drops. EDTA inhibition of growth was considered to be due to suppression of alkane solubilization, brought about by the solubilizing factor(s) produced by cells. It was shown that this chelating agent did not inhibit the growth of yeast on solubilized alkane but strongly inhibited its growth on alkane drops. It was demonstrated that adherent capacity of microbial cell to oil phase was closely related to the state of hydrocarbon emulsification and had no relationship to the ability of organisms to grow on hydrocarbon. Certain proteolytic enzymes inhibited the growth of yeast on alkane, presumably by digesting the alkane solubilizing protein, but not on glucose, and the inhibition was removed by a supply of surfactant-emulsified and surfactant-solubilized alkane. Specific solubilization of various hydrocarbon types during growth of the prokaryotic bacterial strain was demonstrated. The specific solubilization of hydrocarbon was strongly inhibited strain was demonstrated. The specific solubilization of hydrocarbon was strongly inhibited by EDTA, and the inhibition was removed by excess Ca(2+). It was concluded that specific solubilization of hydrocarbons is an important mechanism in the microbial uptake of hydrocarbons.

Anaerobic fluidized bed whey treatment

Abstract: Anaerobic treatment of moderate strength lactic casein why permeate [2000-7000 mg/L soluble chemical oxygen demand (SCOD)] is possible in fluidized bed reactors. Removal efficiencies up to 90% were obtained at organic removal rates of 7.7 kg SCOD M(-3) day(-1) and efficiencies of 70% were obtained at organic removal rates of 19.5 kg SCOD M(-3) day(-1), both at 35 degrees C. A removal rate of 3.0 kg SCOD M(-3) day(-1) at 50% removal efficiency was obtained at 15 degrees C. Nutrient requirements were much lower than for CSTR systems, and no supplemental nitrogen or phosphorus was required. Removal rates increased and removal efficiency decreased as the organic loading increased. Microorganism concentration increased with decreasing temperature, compensating for reduced reaction rates at lower temperatures.

Hydrodynamic stress capacity of microorganisms

Abstract: A new experimental method has been developed for estimating the hydrodynamic stress capacity of microorganisms. In a test apparatus, stable continuous cultures of three types of green algae and two cyanobacteria were exposed to well-defined hydrodynamic loads in a free jet. During and after the stress experiments the cultures showed a different response due to the damage in the jet. The results of these free-jet experiments with short stress exposure were compared to those of stirring experiments in which hydrodynamic load was continuously generated by a stirrer. In both kinds of experiments distinct critical stress values could be determined below which no essential damage of the microorganisms cultures occurred. A correlation between the critical stress values in free-jet and stirring experiments was found. It can be deduced that the free-jet data, expressed as critical volumetric dissipated energy, are suitable for the calculation of hydrodynamic stress to which microorganisms might be exposed in biotechnical plants without suffering damage.

Effects of ethanol and other alkanols on the kinetics and the activation parameters of thermal death in Saccharomyces cerevisiae.

Abstract: Ethanol, isopropanol, propanol, and butanol enhanced thermal death in Saccharomyces cerevisiae by increasing ΔS‡, the entropy of activation of thermal death while ΔH‡, the enthalpy of activation, was not significantly affected. The relation between ΔS‡ and alkanol concentration was linear with a different slope for each alkanol: ΔS‡X = ΔS‡0 + CAEX, where X is the alkanol concentration and CAE the entropy coefficient for the aqueous phase defined as increase in entropy of activation per unit concentrations of the alkanol. CAE was correlated with the lipid-buffer partition coefficients of the alkanols while CME, the entropy coefficient for the lipid phase, was nearly identical for the four alkanol and averaged 37.6 entropy units per mole of alkanol per kilogram of membrane. As predicted by these results, the specific death rates (Kd) at constant temperature were an exponential function of the alkanol concentration and behaved in agreement with the following equation: In KXd = In K0d + (CAE/R)X, where R is the gas constant. It was concluded that the alkanols enhanced thermal death through nonspecific action on membrane structure.