Mixed carbon source utilization of meat-spoiling Pseudomonas fragi 72 in relation to oxygen limitation and carbon dioxide inhibition

The growth of meat-spoiling Pseudomonas fragi 72 was studied on a defined salt medium supplemented with L-aspartate, citrate, creatine, creatinine, D-glucose, L-glutamate, and L-lactate. The utilization of the different carbon sources was followed in batch and continuous culture and under the influence of oxygen limitation and carbon dioxide inhibition (50% CO2 in air). Under nonrestricted atmospheric conditions in batch culture, the organism showed a preference in the utilization of the carbon sources in the order glucose greater than lactate greater than citrate greater than aspartate-glutamate greater than creatine greater than creatinine. The first five sources were utilized simultaneously. The order of preference was changed in continuous culture to lactate-citrate-glutamate-aspartate greater than glucose greater than creatine greater than creatinine. All carbon sources were utilized at lower dilution rates, but as the rate was increased the concentration of the carbon sources started to increase in the effluent and the preference could be seen. Under conditions of oxygen limitation the preference for glucose was weakened, but for lactate it was slightly enhanced (batch and continuous culture). Under conditions of CO2 inhibition, the preference for glucose was enhanced. However, lactate and amino acids were still preferred to glucose in the continuous culture. The utilization of creatine and creatinine was blocked by CO2 in batch culture, and only a slight utilization of creatine was noticed in a chemostat at lower dilution rates.     

Mixed carbon source utilization of meat-spoiling Pseudomonas fragi 72 in relation to oxygen limitation and carbon dioxide inhibition.

The growth of meat-spoiling Pseudomonas fragi 72 was studied on a defined salt medium supplemented with L-aspartate, citrate, creatine, creatinine, D-glucose, L-glutamate, and L-lactate. The utilization of the different carbon sources was followed in batch and continuous culture and under the influence of oxygen limitation and carbon dioxide inhibition (50% CO2 in air). Under nonrestricted atmospheric conditions in batch culture, the organism showed a preference in the utilization of the carbon sources in the order glucose greater than lactate greater than citrate greater than aspartate-glutamate greater than creatine greater than creatinine. The first five sources were utilized simultaneously. The order of preference was changed in continuous culture to lactate-citrate-glutamate-aspartate greater than glucose greater than creatine greater than creatinine. All carbon sources were utilized at lower dilution rates, but as the rate was increased the concentration of the carbon sources started to increase in the effluent and the preference could be seen. Under conditions of oxygen limitation the preference for glucose was weakened, but for lactate it was slightly enhanced (batch and continuous culture). Under conditions of CO2 inhibition, the preference for glucose was enhanced. However, lactate and amino acids were still preferred to glucose in the continuous culture. The utilization of creatine and creatinine was blocked by CO2 in batch culture, and only a slight utilization of creatine was noticed in a chemostat at lower dilution rates.     

Enzymic analysis of the crabtree effect in glucose-limited chemostat cultures of Saccharomyces cerevisiae.

The physiology of Saccharomyces cerevisiae CBS 8066 was studied in glucose-limited chemostat cultures. Below a dilution rate of 0.30 h-1 glucose was completely respired, and biomass and CO2 were the only products formed. Above this dilution rate acetate and pyruvate appeared in the culture fluid, accompanied by disproportional increases in the rates of oxygen consumption and carbon dioxide production. This enhanced respiratory activity was accompanied by a drop in cell yield from 0.50 to 0.47 g (dry weight) g of glucose-1. At a dilution rate of 0.38 h-1 the culture reached its maximal oxidation capacity of 12 mmol of O2 g (dry weight)-1 h-1. A further increase in the dilution rate resulted in aerobic alcoholic fermentation in addition to respiration, accompanied by an additional decrease in cell yield from 0.47 to 0.16 g (dry weight) g of glucose-1. Since the high respiratory activity of the yeast at intermediary dilution rates would allow for full respiratory metabolism of glucose up to dilution rates close to mumax, we conclude that the occurrence of alcoholic fermentation is not primarily due to a limited respiratory capacity. Rather, organic acids produced by the organism may have an uncoupling effect on its respiration. As a result the respiratory activity is enhanced and reaches its maximum at a dilution rate of 0.38 h-1. An attempt was made to interpret the dilution rate-dependent formation of ethanol and acetate in glucose-limited chemostat cultures of S. cerevisiae CBS 8066 as an effect of overflow metabolism at the pyruvate level. Therefore, the activities of pyruvate decarboxylase, NAD+- and NADP+-dependent acetaldehyde dehydrogenases, acetyl coenzyme A (acetyl-CoA) synthetase, and alcohol dehydrogenase were determined in extracts of cells grown at various dilution rates. From the enzyme profiles, substrate affinities, and calculated intracellular pyruvate concentrations, the following conclusions were drawn with respect to product formation of cells growing under glucose limitation. (i) Pyruvate decarboxylase, the key enzyme of alcoholic fermentation, probably already is operative under conditions in which alcoholic fermentation is absent. The acetaldehyde produced by the enzyme is then oxidized via acetaldehyde dehydrogenases and acetyl-CoA synthetase. The acetyl-CoA thus formed is further oxidized in the mitochondria. (ii) Acetate formation results from insufficient activity of acetyl-CoA synthetase, required for the complete oxidation of acetate. Ethanol formation results from insufficient activity of acetaldehyde dehydrogenases.(ABSTRACT TRUNCATED AT 400 WORDS)     

Enzymic analysis of the crabtree effect in glucose-limited chemostat cultures of Saccharomyces cerevisiae

The physiology of Saccharomyces cerevisiae CBS 8066 was studied in glucose-limited chemostat cultures. Below a dilution rate of 0.30 h-1 glucose was completely respired, and biomass and CO2 were the only products formed. Above this dilution rate acetate and pyruvate appeared in the culture fluid, accompanied by disproportional increases in the rates of oxygen consumption and carbon dioxide production. This enhanced respiratory activity was accompanied by a drop in cell yield from 0.50 to 0.47 g (dry weight) g of glucose-1. At a dilution rate of 0.38 h-1 the culture reached its maximal oxidation capacity of 12 mmol of O2 g (dry weight)-1 h-1. A further increase in the dilution rate resulted in aerobic alcoholic fermentation in addition to respiration, accompanied by an additional decrease in cell yield from 0.47 to 0.16 g (dry weight) g of glucose-1. Since the high respiratory activity of the yeast at intermediary dilution rates would allow for full respiratory metabolism of glucose up to dilution rates close to mumax, we conclude that the occurrence of alcoholic fermentation is not primarily due to a limited respiratory capacity. Rather, organic acids produced by the organism may have an uncoupling effect on its respiration. As a result the respiratory activity is enhanced and reaches its maximum at a dilution rate of 0.38 h-1. An attempt was made to interpret the dilution rate-dependent formation of ethanol and acetate in glucose-limited chemostat cultures of S. cerevisiae CBS 8066 as an effect of overflow metabolism at the pyruvate level. Therefore, the activities of pyruvate decarboxylase, NAD+- and NADP+-dependent acetaldehyde dehydrogenases, acetyl coenzyme A (acetyl-CoA) synthetase, and alcohol dehydrogenase were determined in extracts of cells grown at various dilution rates. From the enzyme profiles, substrate affinities, and calculated intracellular pyruvate concentrations, the following conclusions were drawn with respect to product formation of cells growing under glucose limitation. (i) Pyruvate decarboxylase, the key enzyme of alcoholic fermentation, probably already is operative under conditions in which alcoholic fermentation is absent. The acetaldehyde produced by the enzyme is then oxidized via acetaldehyde dehydrogenases and acetyl-CoA synthetase. The acetyl-CoA thus formed is further oxidized in the mitochondria. (ii) Acetate formation results from insufficient activity of acetyl-CoA synthetase, required for the complete oxidation of acetate. Ethanol formation results from insufficient activity of acetaldehyde dehydrogenases.(ABSTRACT TRUNCATED AT 400 WORDS)     

The regulatory effects of growth rate and cyclic AMP levels on carbon catabolism and respiration in Escherichia coli K-12.

Cyclic AMP levels in glucose and succinate-limited and ammonia-limited glucose-containing continuous cultures of Escherichia coli were measured at different bacterial growth rates. Intracellular cyclic AMP concentrations were fairly constant (about 5 micrometer) at all dilution rates used when glucose was limiting. In ammonia-limited glucose cultures the cyclic AMP content was much lower (about 0.3 micrometer). In succinate-limited cultures cyclic AMP levels fell from 2.7 to 0.8 micrometer as dilution rate increased from 0.05 to 0.4 h-1. The effects of cyclic AMP on respiratory and carbon catabolic enzyme levels were studied. There was no indication of a direct cyclic AMP involvement in the regulation of these cellular functions. It seems more likely that the variations in enzyme levels observed resulted from variation of the specific growth rate of cultures.     

Metabolic and energetic control of Pseudomonas mendocina growth during transitions from aerobic to oxygen-limited conditions in chemostat cultures.

Several metabolic fluxes were analyzed during gradual transitions from aerobic to oxygen-limited conditions in chemostat cultures of Pseudomonas mendocina growing in synthetic medium at a dilution rate of 0.25 h-1. P. mendocina growth was glucose limited at high oxygen partial pressures (70 and 20% pO2) and exhibited an oxidative type of metabolism characterized by respiratory quotient (RQ) values of 1.0. A similar RQ value was obtained at low pO2 (2%), and detectable levels of acetic, formic, and lactic acids were determined in the extracellular medium. RQs of 0.9 +/- 0.12 were found at 70% pO2 for growth rates ranging from 0.025 to 0.5 h-1. At high pO2, the control coefficients of oxygen on catabolic fluxes were 0.19 and 0.22 for O2 uptake and CO2 production, respectively. At low pO2 (2%), the catabolic and anabolic fluxes were highly controlled by oxygen. P. mendocina showed a mixed-type fermentative metabolism when nitrogen was flushed into chemostat cultures. Ethanol and acetic, lactic, and formic acids were excreted and represented 7.5% of the total carbon recovered. Approximately 50% of the carbon was found as uronic acids in the extracellular medium. Physiological studies were performed under microaerophilic conditions (nitrogen flushing) in continuous cultures for a wide range of growth rates (0.03 to 0.5 h-1). A cell population, able to exhibit a near-maximum theoretical yield of ATP (YmaxATP = 25 g/mol) with a number of ATP molecules formed during the transfer of an electron towards oxygen along the respiration chain (P/O ratio) of 3, appears to have adapted to microaerophilic conditions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Metabolic and energetic control of Pseudomonas mendocina growth during transitions from aerobic to oxygen-limited conditions in chemostat cultures.

Several metabolic fluxes were analyzed during gradual transitions from aerobic to oxygen-limited conditions in chemostat cultures of Pseudomonas mendocina growing in synthetic medium at a dilution rate of 0.25 h-1. P. mendocina growth was glucose limited at high oxygen partial pressures (70 and 20% pO2) and exhibited an oxidative type of metabolism characterized by respiratory quotient (RQ) values of 1.0. A similar RQ value was obtained at low pO2 (2%), and detectable levels of acetic, formic, and lactic acids were determined in the extracellular medium. RQs of 0.9 +/- 0.12 were found at 70% pO2 for growth rates ranging from 0.025 to 0.5 h-1. At high pO2, the control coefficients of oxygen on catabolic fluxes were 0.19 and 0.22 for O2 uptake and CO2 production, respectively. At low pO2 (2%), the catabolic and anabolic fluxes were highly controlled by oxygen. P. mendocina showed a mixed-type fermentative metabolism when nitrogen was flushed into chemostat cultures. Ethanol and acetic, lactic, and formic acids were excreted and represented 7.5% of the total carbon recovered. Approximately 50% of the carbon was found as uronic acids in the extracellular medium. Physiological studies were performed under microaerophilic conditions (nitrogen flushing) in continuous cultures for a wide range of growth rates (0.03 to 0.5 h-1). A cell population, able to exhibit a near-maximum theoretical yield of ATP (YmaxATP = 25 g/mol) with a number of ATP molecules formed during the transfer of an electron towards oxygen along the respiration chain (P/O ratio) of 3, appears to have adapted to microaerophilic conditions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Large-scale (20 l) culture of surface-immobilized Catharanthus roseus cells.

Surface-immobilized C. roseus cell cultures were grown in a 20-l modified airlift bioreactor operated at 0.51 vvm (kLa approximately 8 h-1) under various gassing regimes [air, 2% (v/v) and 5% CO2]. Extracellular ammonium, phosphate, and nitrate ions as well as carbohydrate uptake and pH value of the medium were monitored together with on-line dissolved oxygen concentration, conductivity of the medium, and carbon dioxide production rate (CPR) of the cultures. Cultures supplemented with 2% CO2 showed higher nitrate (5.0-7.0 mM d-1) and carbohydrate (3.3 g l-1 d-1) uptake rates and biomass production (mu approximately 0.24 d-1, yield approximately 0.33 g dw g CHO-1 and 7.4 g dw L-1) as compared to air (3.6 mM d-1, 2.1 g l-1 d-1; 0.20 d-1, 0.25 g dw g CHO-1 and 5 g dw l-1) and 5% CO2 (2.0-3.6 mM d-1, 2.0 g l-1 d-1; 0.11 d-1, 0.20 g dw g CHO-1 and 5 g dw l-1) cultures and as reported previously for suspension cultures. In addition, air and 5% CO2 cultures displayed incomplete carbohydrate uptake and, more important, phosphate and ammonium ion release into the medium at the end, which was ascribed to loss of viability. This was not observed for 2% CO2 immobilized bioreactor as well as shake flask control suspension cultures, which suggests that sparged C. roseus surface-immobilized cell cultures require 2% CO2 supplementation of the gas phase for both maximum growth and retained viability. The maximum CPRs of all cultures were in the same range (2.1-2.8 mM CO2 l-1 h-1). However, the estimated maximum specific CO2 production rates of 2% CO2 and 5% CO2 immobilized cultures (0.6 mM g dw-1 h-1) were lower than those found for air-sparged immobilized cultures (1.0-1.3 mM g dw-1 h-1). These rates are significantly higher than those reported in the literature for C. roseus cell suspension cultures performed in bioreactors gassed with air (approximately 0.2-0.55 mM g dw-1 h-1).     

Biosynthesis of exopolysaccharide by Pseudomonas aeruginosa.

In batch cultures of Pseudomonas aeruginosa, the maximum rate of exopolysaccharide synthesis occurred during exponential growth. In nitrogen-limited continuous culture, the specific rate of exopolysaccharide synthesis increased from 0.27 g g of cell-1 h-1 at a dilution rate (D) of 0.05 h-1 to 0.44 g g of cells h-1 at D=0.1 H-1. The yield of exopolysaccharide on the basis of glucose used was in the range of 56 to 64%. Exopolysaccharide was also synthesized in carbon-limited cultures at 0.19 g g of cell-1 h-1 at D=0.05 h-1 in a 33% yield. Nonmucoid variants appeared after seven generations in continuous culture and rapidly increased in proportion to the total number of organisms present.     

Cultivation of Escherichia coli with mixtures of 3-phenylpropionic acid and glucose: steady-state growth kinetics.

The fate of pollutants in the environment is affected by the presence of easily degradable carbon sources. As a step towards understanding these complex interactions, a model system was explored: the degradation of mixtures of glucose (i.e., an easily degradable substrate) and 3-phenylpropionic acid (3ppa) (a model pollutant) by Escherichia coli ML 30 was studied systematically in carbon-limited continuous culture. The two substrates were always consumed simultaneously regardless of the dilution rate applied. Even at dilution rates higher than the maximum specific growth rate for 3ppa (0.35 +/- 0.05 h-1), the two carbon substrates were utilized together. When cells were grown at a constant dilution rate with different mixtures of 3ppa and glucose, in which 3ppa contributed between 5 and 90% of carbon substrate in the feed medium, the steady-state concentrations of 3ppa and glucose were approximately proportional to the ratio of the two substrates in the feed medium. When cells were cultivated at different dilution rates with a 1:1 mixture (based on carbon) of glucose and 3ppa, an overall maximum specific growth rate of 0.90 +/- 0.05 h-1 and a Monod substrate saturation constant for 3ppa (Ks) of 600 to 700 micrograms liter-1, similar to that measured during growth with 3ppa alone, fitted the experimentally determined steady-state 3ppa concentrations. However, due to the highly differing substrate affinity constants for 3ppa and glucose (Ks approximately 30 to 70 micrograms liter-1), the total steady-state carbon concentration in the culture at a constant dilution rate was determined mainly by the steady-state 3ppa carbon concentration, and it increased with increasing proportions of 3ppa in the feed medium.     

Effect of oxygen supply on growth ofKlebsiella aerogenes in a multistage tower fermentor

The effect of the rate of oxygen supply on biomass growth, consumption of carbon source formation of metabolic by-products, biomass yeilds referred to C-source and oxygen, respiration rate and the respiratory quotient was studied in a multistage tower fermentor with an interstage backflow, i.e. with a continuous reinoculation of the preceding stages. Experiments were done with Klebsiella aerogenes CCM 2318 in a synthetic glucose medium with constant glucose concentration in the feed, at pH 7.0. temperature 30 degrees C, and dilution rates 0.6 and 0.178 h-1 (referred to one stage). Different behavior of the culture was found at different dilution rates both with oxygen and under oxygen limitation. As compared with the chemostat system, the regime with an interstage backflow exhibited differences in respiration rate and CO2 formation; this attests to a considerably different physiological state of the cells.     

Defining an optimal carbon source/methionine feed strategy for growth and cephalosporin C formation by Cephalosporium acremonium.

The effect of the method of methionine addition, growth-limiting carbon source (glucose vs sucrose), and culture growth rate on cephalosporin C production was investigated in a Cephalosporium acremonium defined medium fed batch fermentation. Batch addition of methionine, at a concentration of 3 g/L, prior to the start of a fed sucrose fermentation was found to interfere with the ability of the culture to utilize this sugar, thus limiting growth and decreasing cephalosporin C production. Batch methionine addition had no effect on glucose-limited cultures. Concurrent exponential feeding of methionine with sucrose improved both culture growth and productivity. Under the control of identical carbon source limiting feed profiles, sucrose was observed to support greater cephalosporin C production than glucose. Optimal cephalosporin C production in a C. acremonium defined medium fed batch fermentation was obtained through controlling culture growth during the rapid growth phase at a relatively low level with respect to mumax (mu approximately 0.036 h-1) until achieving a desired cell mass with a concurrent sucrose and methionine feed, followed by maintaining relatively vigorous growth (mu approximately 0.01 h-1) with sucrose for the duration of the fermentation.     

Continuous hydrogen production by the hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1

The hydrogen (H2) production potential of the hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1 was evaluated at 85 degrees C. In batch cultivation using a complex medium supplemented with elemental sulfur (S0), evolution of H2S and CO2 was observed in the gas phase. When S0 was omitted and pyruvate or starch was added in the medium, the cells produced H2 at high levels instead of H2S. As the level of H2 appeared to correlate with the specific growth rate, analysis in continuous cultures was performed to develop a continuous H2 production system. In a steady-state condition at a dilution rate of 0.2 h-1, a continuous H2 production rate (per gram dry weight, gdw) of 24.9 and 14.0 mmol gdw-1 h-1 was observed in media supplemented with pyruvate and starch, respectively. In both cultivations, a high accumulation of acetate and alanine was found as metabolites. When the dilution rates were elevated in the medium with pyruvate, steady-state growth was observed up to 0.8 h-1, and a maximum H2 production rate of 59.6 mmol gdw-1 h-1 was obtained. Based on the experimental results along with data of the entire genome sequence, the metabolic pathway of the strain relating to starch and pyruvate degradation is discussed.     

Regulation of beta-D-galactosidase synthesis in Candida pseudotropicalis.

Regulation of lactose (beta-D-galactosidase) synthesis in the lactose-utilizing yeast Candida pseudotropicalis was studied. The enzyme was inducible by lactose and galactose. When grown on these sugars the enzyme level of the yeast was 20 times or higher than when grown on glycerol. The Km and optimal pH were similar for the lactase induced either by lactose or galactose. The hydrolysis of o-nitrophenyl-beta-D-galactopyranoside by the lactase was inhibited by galactose and several analogs and galactosides, but not by glucose. Lactose uptake activity observed in lactose-grown cells was very reduced in cells grown on glucose or galactose. Glucose repressed the induction of lactase, but not the metabolic system for galactose utilization. In continuous culture on lactose medium at dilution rates below 0.2 h-1 the specific lactase activity was higher than in batch cultures and decreased with increases in dilution rate. Lactase was induced by pulses of lactose and galactose in cells growing on glucose, but only at low dilution rates were the steady-state concentration of glucose was very low.     

Effect of yeast growth conditions on yeast-mycelial transition in Candida albicans

When grown and induced to form germ tubes in liquid defined media, yeast cells of Candida albicans must reach stationary phase before acquiring ability to carry out the yeast-mycelial transition. This study examined the effect of the carbon source utilized for yeast growth on the inducibility of stationary phase yeast. When grown to the same stationary phase cell density as glucose cultures, cultures grown on citrate were fully inducible while cultures grown on galactose and mannose showed a small reduction. Cultures grown on ethanol were reduced 80% in morphological conversion. When glucose grown cells were induced in the presence of these carbon sources, hexoses supported full induction while ethanol reduced induction 80%. Induction in the presence of carboxylic acids was similar to induction in the absence of added carbon source. When induced on the same source used in yeast growth, germ tube formation was reduced for all carbon sources except hexoses. When induced in the absence of added carbon source, yeasts grown on citrate and ethanol were inhibited 80-100%. Cultures starved for glucose were more inhibited than cultures starved for NH4Cl when induced without added carbon source. These observations suggest that the metabolic state of the stationary phase cell is an important factor in the ability to respond to conditions inducing germ tube formation.     

Carbon dioxide assimilation by Thiobacillus novellus under nutrient-limited mixotrophic conditions

The contribution of CO2 to cell material synthesis in Thiobacillus novellus under nutrient-limited conditions was estimated by comparing 14CO2 uptake rates of steady-state autotrophic cultures with that of heterotrophic and mixotrophic cultures at a given dilution rate. Under heterotrophic conditions, some 13% of the cell carbon was derived from CO2; this is similar to the usual anaplerotic CO2 fixation in batch cultures of heterotrophic bacteria. Under mixotrophic conditions, the contribution of CO2 to cell material synthesis increased with increasing S2O3 2- -to-glucose ratio in the medium inflow; at a ratio of 10, ca. 32% of the cell carbon was synthesized from CO2. We speculate that the use of CO2 as carbon source, even when the glucose provided is sufficient to fulfill the biosynthetic needs, may augment the growth rate of the bacterium under such nutrient-limited conditions and could therefore be of survival value in nature. Some of the CO2 assimilated was excreted into the medium as organic compounds under all growth conditions, but in large amounts only in autotrophic environments as very low dilution rates.     

Some observations on protease production in continuous suspension cultures of Bacillus firmus

Invariance of culture conditions in steady state continuous cultures make these a very valuable tool to study the influence of various culture parameters on cell growth and synthesis of primary and secondary metabolites. The result of a parametric study on production of protease in continuous suspension cultures of Bacillus firmus NRS 783 are reported in this article. This strain is a superior producer of an alkaline protease with major application in the detergent industry. The parameters investigated include dilution rate and concentrations of yeast extract, ammonium, and inorganic phosphate in the bioreactor feed, glucose being the principal carbon source in all experiments. The regulatory effects of the key culture parameters on cell growth, synthesis and secretion of protease, and production of acetic acid are investigated. The relations among the specific cell growth rate, specific utilization rates of the principal carbon, nitrogen, and phosphorous sources, and specific production rates of two nonbiomass products, viz., acetic acid and protease, are examined, and the effects of the manipulated culture parameters on these relations, specific protease activity, and yields of cell mass, protease, and acetic acid on the basis of the principal carbon, nitrogen, and phosphorous sources are studied. An increase in dilution rate led to increases in specific utilization rates of the principal carbon, nitrogen, and phosphorous sources and specific production rates of acetic acid and protease and decreases in bulk activities/concentrations of the three products (acetic acid, cell mass, and protease). As a result, the productivities of the three species were maximized at an intermediate dilution rate. Increased supply of yeast extract (a rich source of amino acids, proteins, and vitamins, besides being an additional source of carbon, nitrogen, and phosphorus) promoted cell mass formation but reduced protease production per unit cell mass. Increased supply of nitrogen and phosphorous sources stimulated protease synthesis up to certain threshold levels and repressed the enzyme synthesis beyond the threshold levels. With increased supply of the nitrogen source, the phosphorous source was more efficiently utilized for cell growth and protease synthesis. Stable maintenance of continuous cultures of B. firmus over prolonged period is demonstrated in this study. (c) 1993 John Wiley & Sons, Inc.     

The influence of different carbon sources and medium osmolarity on the potassium requirements of Candida utilis NCYC 321, growing in continuous culture

In order to study the influence of different carbon sources on the K⁺-requirements of Candida utilis NCYC 321, this yeast was grown at several different dilution rates in potassium-limited continuous cultures with either glucose, glycerol, ethanol, citrate or lactate serving as the carbon and energy source.It was found that the nature of the carbon source profoundly influenced the cellular potassium content, especially at low dilution rates, but that these differences could not be correlated with any differences in relative growth rate (i.e., / _max. And although small amounts of potassium seemingly were needed to serve in osmoregulation and in the cotransport of some acidic carbon sources (lactate and citrate), these requirements were negligible.Independent of carbon source, a strong correlation existed between the intracellular potassium concentration and the yield value on oxygen (Y _O), and between cellular potassium concentration and growth rate. From these two correlations it was concluded that potassium probably was involved mainly in processes associated with ATP synthesis in this yeast.Finally the effect of the addition of NaCl to the medium was tested with glucose-containing cultures that were either carbon- or potassium-limited. Up to a concentration of 20 g/l, NaCl was without influence on Y _O, Y _glucose and q _{O 2}, but effected a slight increase in the cellular potassium content of the potassium-limited cells and a decrease in that of the glucose-limited cells.