Lysine production from hydrocarbon by micrococcus species

A bacterium isolated from Assam (India) soil was found to accumulate L-lysine in the mineral salt-hydrocarbon medium and identified to be a strain of Micrococcus luteus. The strain is able to grow and accumulate l-lysine in a purely synthetic medium, but supplementation of the synthetic medium with casamino acid or yeast extract or both, improves the yield. The entire fermentation period can be divided into a growth phase and a production phase, which can be prolonged by adjustment of the pH to the neutral range. Among the different hydrocarbon and nitrogen sources tested straight run gas oil (47percnt;) and ammonium sulphate (0.4%), respectively, were found most suitable. Erythromycin at 1 μg/ml level inhibited growth bu¸t stimulated lysine excretion. An inoculum level of 10% (v/v) of the medium was optimal for lysine production. The yield of lysine under optimal conditions was found to be 3.25 g per litre of the medium. Lysine has been isolated in crystalline form from the fermented broth by ion exchange resin chromatography and found to be pure sample of L-isomer.     

Valine production from hydrocarbon byMicrococcus varians

A bacterium isolated from Assam (India) soil was found to accumulateL-valine in the growth medium and was identified asMicrococcus varians. The strain grew and accumulated valine in a purely synthetic medium, but supplementation with either casamino acids or yeast extract or with both, improved the yield. The entire fermentation period could be divided into a growth phase and a production (phase which could be prolonged by adjustment of pH to neutral range. Among the different hydrocarbon and nitrogen sources tested straight run gas-oil and ammonium sulphate, respectively, were found most suitable. Antibiotics inhibited growth but stimulated extracellular valine accumulation. Vitamins stimulated growth and valine yield and an inoculum level of 10 % was found to be optimal. The yield ofL-valine under optimal conditions was 2.95 g/L.     

Extracellular lysine production from hydrocarbons byArthrobacter globiformis

A bacterium isolated from Burdwan (India) soil was found to accumulatel-lysine in the growth medium and was identified asArthrobacter globiformis. The strain grew and accumulated lysine in a purely synthetic medium. Supplementation of the synthetic medium stimulated growth but did not improve the yield. The entire fermentation period could be divided into a growth phase and a production phase, which could be prolonged by adjustment of pH to the neutral range. Among the different hydrocarbon and nitrogen sources tested SR gas oil at 4 % and ammonium sulphate at 0.4 %, respectively, were found most to be suitable. Different vitamins and antibiotics stimulated growth and lysine yield; inoculum of 7 % (V/V) of the medium was found to be optimal. The yield of lysine under optimal conditions was 3.4 g per litre medium. Lysine was isolated in crystalline form from the fermented broth by IEC and found to be a purel-isomer.     

Production of l-glutamic acid by aBacillus sP

A strain ofBacillus cereus var.mycoides isolated from Burdwan soil producesl-glutamate in the medium. The strain is able to grow and produce in a synthetic medium but supplementation with casamino acid or yeast extract improves the yield. Maintenance of pH of the fermentation medium near neutrality prolongs the active growth period and improves the yield. Glucose and ammonium nitrate were found to be most suitable carbon and nitrogen sources, respectively. Cane sugar molasses (as a substitute for glucose) significantly stimulated the growth but glutamate production was less. Various B vitamins stimulate the growth and glutamate yield. The yield of glutamate under optimal condition is 5.2 g/l.     

Production of glutamic acid byArthrobacter globiformis: Influence of cultural conditions

Arthrobacter globiformis isolated from Burdwan soil excretes glutamic acid in a glucose mineral salt medium with suboptimal level of biotin. Glutamate begins to accumulate in the medium from the mid-exponential phase of growth and its excretion could be prolonged by adjustment of pH to the neutral range. Among the different carbon and nitrogen sources tested glucose (8%) and ammonium nitrate (0.53%), respectively, were found to be most suitable. Molasses could not be used as a substitute for glucose even if antibiotics or Tween 80 are incorporated in the medium. Bacitracin (1 μg/mL) stimulated glutamate excretion. Atemperature of 28°C and an inoculum dose of 4% were optimal for production. Under optimal conditions, in the flask culture the isolate excreted 16.1 g glutamic acid per litre in 120 h. Glutamic acid isolated from the fermented broth was found to be purel-diastereomer.     

Lysine production from hydrocarbon by Micrococcus varians 2Fa

A bacterium isolated from Assam (India) soil was found to accumulale l-lysine from hydrocarbon and was identified as a strain of Micrococcus varians. The strain is able to grow and accumulate lysine in a purely synthetic medium though supplementation of the synthetic medium with casamino acids significantly improves the yield. The yield of l-lysine under optimal conditions was found to be 2.6 g X 1(-1) of the compound isolated in crystalline form.     

Formation of Valine in the Presence of Sodium Acrylate Monomer by Glutamic Acid Producing Bacteria

Glutamic acid producing bacteria accumulated a large amount of valine in the presence of the excess biotin, when sodium acrylate monomer (Na-AM) was added at the earlier phase of culture. Brevibacterium roseum ATCC 13825, particularly, accumulated the large amount of valine among bacteria tested and the conditions of valine accumulation by this strain were investigated.

The most effective addition time of Na-AM was at the earlier phase of logarithmic phase. The optimal concentration of Na-AM for the accumulation of valine was 1.0 per cent (v/v). Most effective nitrogen sources were the combination of 1.0 per cent urea and 0.2 per cent ammonium sulphate. The additions of Mn²⁺ and Fe²⁺ increased valine accumulation. By the excess concentration of biotin for growth, 20 μg/liter or more, did not affected valine accumulation, while the presence of the suboptimal condition of biotin for growth was not good for the formation of valine even in the presence of Na-AM. The accumulation of valine reached 9.0 mg/ml from 75.0 mg/ml of glucose in the presence of 50 μg/liter of biotin and 1.0 per cent (v/v) of Na-AM.

This strain possessed considerable activity of valine formation regardless of the addition of Na-AM and promoted the accumulation of valine by the addition of Na-AM.     

Microbial hydrogen production with Bacillus coagulans IIT-BT S1 isolated from anaerobic sewage sludge

Bacillus coagulans strain IIT-BT S1 isolated from anaerobically digested activated sewage sludge was investigated for its ability to produce H(2) from glucose-based medium under the influence of different environmental parameters. At mid-exponential phase of cell growth, H(2) production initiated and reached maximum production rate in the stationary phase. The maximal H(2) yield (2.28 mol H(2)/molglucose) was recorded at an initial glucose concentration of 2% (w/v), pH 6.5, temperature 37 degrees C, inoculum volume of 10% (v/v) and inoculum age of 14 h. Cell growth rate and rate of hydrogen production decreased when glucose concentration was elevated above 2% w/v, indicating substrate inhibition. The ability of the organism to utilize various carbon sources for H(2) fermentation was also determined.     

Nutritional Requirements of Staphylococcus aureus S-6

A synthetic medium was devised for growth of Staphylococcus aureus strain S-6. The growth yield in synthetic medium was compared to that in complex medium containing an equivalent amount of protein hydrolysate. Enterotoxin B formation in the two media was also compared. The defined medium was composed of inorganic salts, 11 amino acids (glycine, valine, leucine, threonine, phenylalanine, tyrosine, cysteine, methionine, proline, arginine, and histidine), and three vitamins (thiamine, nicotinic acid, and biotin). Biotin was a growth factor requirement of S-6 when glutamic acid but not glucose was used as a carbon source. The quantity of enterotoxin B produced in the defined medium was about one-seventh of that produced in complex medium, even though the growth yields were similar.     

Characterization of optimal growth conditions and carotenoid production of strain Rhodotorula mucilaginosa HP isolated from larvae of Pieris rapae

Yeasts have been studied because of their production of a pigment known as carotenoid with potential application in food and feed supplements. A carotenoid‐producing yeast was isolated from the larvae of Pieris rapae, named HP. The strain HP was identified as Rhodotorula mucilaginosa classified by its carbohydrate fermentation pattern and physiological tests. Rhodotorula mucilaginosa HP produces several exogenous enzymes: alkaline phosphatase, esterase, leucine arylamidase, valine arylamidase, acid phosphatase and β‐glucosidase. Using response surface methodology, selected medium components (yeast extract, malt extract, peptone, glucose) were tested to find the optimum conditions for carotenoid production and the growth of R. mucilaginosa HP. Central composite design was used to control the concentrations of medium components. Peptone and glucose had the largest effects on carotenoid production and cell growth of R. mucilaginosa HP, respectively. The estimated optimal growth conditions of R. mucilaginosa HP were: yeast extract 3.23%, malt extract 2.84%, peptone 6.99% and glucose 12.86%. The estimated optimal conditions for carotenoid production were: yeast extract 2.17%, malt extract 2.11%, peptone 5.79% and glucose 12.46%. These results will assist in the formulation of an appropriate culture medium for optimal carotenoid production of R. mucilaginosa HP for commercial use.     

Influence of antibiotics and trace salts on lysine production by arthrobacter globiformis

A hydrocarbon utilizing strain of Arthrobacter globiformis Lb isolated from local soil has been found to yield lysine 3.4 g l^?1, keeping the medium optimal for pH, C- and N-sources. Addition of antibiotics and micronutrients to that optimal media stimulated cell growth and enhanced lysine yield.     

Production of l-phenylalanine by double auxotrophic mutants of arthrobacter globiformis: Optimization of c and n-source

A number of tryptophan plus tyrosine double auxotrophic mutants isolated by NTG treatment of a glutamic acid prodcing strain of A. globiformis were found to excrete phenylalanine alone in glucose, ammonium chloride, mineral salt medium. Among 24 different carbon sources tested glucose was found to be the best and optimum for -phenylalanine production was at 350 mM level. Among 14 different inorganic and organic nitrogen sources tested ammonium chloride was the best and was optimal at 60 mM level for phenylalanine production. On the optimum level of C and N sources a yield of 1.5 g/l phenylalanine was achieved.     

Arabitol accumulation in Geotrichum candidum

Culture conditions which lead to the intracellular accumulation of arabitol and mannitol in Geotrichum candidum were investigated. The accumulation of arabitol was dependent on the concentrations of metabolizable hexoses, the non-metabolizable disaccharide sucrose, NaCl and KCl in the growth medium. In media containing 2% (w/v) glucose, fructose or l-sorbose cultures contained only mannitol after 48 h or 72 h growth. In media containing 10% (w/v) to 30% (w/v) glucose, or 25% (w/v) fructose or l-sorbose there was an increase in the total concentration of intracellular polyol due to the accumulation of arabitol. This pentitol was also found to accumulate intracellularly when the organism was grown in medium containing 34% (w/v) sucrose, 0.7 M NaCl or 0.7 M KCl in addition to 2% (w/v) glucose. Under the conditions tested no change in the accumulation of mannitol or ethanol-soluble carbohydrate, believed to be primarily composed of trehalose, was evident.Intracellular polyol was released during incubation of arthrospores obtained from media containing 25% or 10% glucose, in distilled water at 25° C, but no polyol was released under these conditions from arthrospores obtained from growth in 2% glucose medium.     

Biosynthesis of amino acids from hydrocarbons

Optimum conditions for the growth ofPseudomonas arvilla, a hydrocarbon utiliser, have been studied. The microorganism produced economic cell yield at pH 5.7 and 4% kerosene concentration. C₁₀-C₁₆ hydrocarbons were utilised by the strain. The growth was maximum on n-decane. Supplementation of the hydrocarbon medium with 0.5% glucose stimulated the growth. Glutamic acid 16.0 mg; leucine 9.0 mg; valine 10.0 mg; methionine 2.5 mg; arginine 2.5 mg; histidine 1.0 mg were present in 100 ml of the broth. Cell protein contained leucine 13.69%, isoleucine 4.9%, histidine 4.37%, tryptophan 2.33%, methionine 1.8% and arginine 2.70%.     

Occurrence of Poly-β-Hydroxybutyrate in the Azotobacteriaceae

Poly-beta-hydroxybutyrate (PHB) from various representative strains of the genera Azotobacter, Beijerinckia, and Derxia was isolated and characterized. During growth in shake culture, with glucose as a carbon and energy source, and molecular nitrogen as a nitrogen source, increase in dry weight appeared linear, and PHB formed a constant percentage of the dry weight. In a medium containing 1% (w/v) glucose, PHB declined with the onset of the stationary phase of growth; with 2% (w/v) glucose, an increase in PHB content during stationary phase was noted in the case of some strains, before a subsequent decline. The decrease in PHB as a percentage of dry cellular weight (not of total amount present in the culture) during growth of some strains with 2% as opposed to 1% (w/v) glucose may be ascribed to a greater production of capsular polysaccharide. PHB content could not be used as a taxonomic criterion. Strain differences were as great as or greater than species differences. The only strain of Beijerinckia fluminensis obtained contained PHB, but it could not be grown on the nitrogen-free medium used. Two species of the genus Azotomonas, reported to be aerobic, nonsymbiotic nitrogen-fixers, did not grow on the nitrogen-free medium used and did not produce PHB during growth with a combined nitrogen source.     

Studies on the Accumulation of Orotic Acid by Escherichia coli K12

More than 300 mg/liter of orotic acid was found to accumulate in the supernatants of the cultures of wild type strains of E. coli K12. The pyrimidine precursor was accumulated in a synthetic medium such as glucose-ammonium sulfate medium. The substance was isolated from the culture, crystallized, and identified as orotic acid. Orotic acid was excreted mainly during logarithmic phase of the bacterial growth. Yeast extract or nutrient broth stimulated bacterial growth, but suppressed orotic acid accumulation. E. coli strains other than K12 failed to accumulate orotic acid.

The results suggest that the accumulation of orotic acid is specific to E. coli K12.     

Isolation and characterization of an anaerobic dehydrodivanillin-degrading bacterium

A novel, strictly anaerobic, gram-negative, non-spore-forming, fusiform, rod-shaped bacterium having high dehydrodivanillin (DDV)-degrading activity was isolated from cow ruminal fluid. This strain degraded a range of six main lignin-related compounds such as DDV, ferulic acid, dehydrodiisoeugenol, guaiacoxyacetic acid, vanillin, and veratrylglycerol-beta-guaiacyl ether to the extent of 14 to 83% within 2 days under strictly anaerobic conditions. As DDV degradation intermediates, three aromatic compounds (dehydrodivanillic acid, vanillic acid, and 5-carboxyvanillic acid) and two alicyclic compounds (cyclohexanecarboxylic acid and cyclohexanol) were detected by thin-layer, high-performance liquid, and gas chromatography and mass spectrometry. The addition of 1% glucose and peptone in a synthetic medium stimulated growth of the strain but slowed down DDV degradation. The presence of 0.1% yeast extract increased both cell growth and DDV degradation. The growth yield in defined medium was 151.5 g (dry weight) of cells per mol of DDV utilized. Characterization of the strain indicated that it was distinct from known Fusobacterium and Clostridium species. The bacterium was easily induced to form protoplasts after treatment with either penicillin or lysozyme. The frequencies of protoplast formation and regeneration in the strain were 94 and 18%, respectively.     

Isolation and characterization of an anaerobic dehydrodivanillin-degrading bacterium.

A novel, strictly anaerobic, gram-negative, non-spore-forming, fusiform, rod-shaped bacterium having high dehydrodivanillin (DDV)-degrading activity was isolated from cow ruminal fluid. This strain degraded a range of six main lignin-related compounds such as DDV, ferulic acid, dehydrodiisoeugenol, guaiacoxyacetic acid, vanillin, and veratrylglycerol-beta-guaiacyl ether to the extent of 14 to 83% within 2 days under strictly anaerobic conditions. As DDV degradation intermediates, three aromatic compounds (dehydrodivanillic acid, vanillic acid, and 5-carboxyvanillic acid) and two alicyclic compounds (cyclohexanecarboxylic acid and cyclohexanol) were detected by thin-layer, high-performance liquid, and gas chromatography and mass spectrometry. The addition of 1% glucose and peptone in a synthetic medium stimulated growth of the strain but slowed down DDV degradation. The presence of 0.1% yeast extract increased both cell growth and DDV degradation. The growth yield in defined medium was 151.5 g (dry weight) of cells per mol of DDV utilized. Characterization of the strain indicated that it was distinct from known Fusobacterium and Clostridium species. The bacterium was easily induced to form protoplasts after treatment with either penicillin or lysozyme. The frequencies of protoplast formation and regeneration in the strain were 94 and 18%, respectively.     

Some aspects of the regulation of the production of extracellular proteolytic enzymes by a marine bacterium

A highly proteolytic Gram-negative, rod-shaped bacterium was isolated from the gills of fresh plaice and the effect of culture conditions on the production of proteolytic enzymes was investigated. When the organism, strain SA 1, was grown in the presence of complex mixtures of proteins and amino acids, both endopeptidase and aminopeptidase activity was demonstrated in the cell-free culture medium. However, synthesis of these enzymes was not observed when the organism was grown in a mineral medium with lactate or succinate as the only carbon and energy source. Synthesis of both endopeptidase and aminopeptidase was induced by the presence of amino acids in the medium. Of the amino acids tested, l-phenylalanine was found to be the best single inducer for the production of endopeptidase. When in addition one or more different amino acids were added, endopeptidase production was found to increase with increasing complexity of the mixture, up to a maximum which was obtained with five different amino acids. Production of the aminopeptidase was optimal when l-glutamic acid was used as a single inducer. For this enzyme the amount of enzyme activity released in the medium decreased with increasing complexity of the amino acid mixture. Endopeptidase as well as aminopeptidase activity was found to accumulate in the medium at the end of the logarithmic growth phase, when the culture was no longer growing exponentially. When the stationary phase was reached, enzyme production stopped. Production of both enzymes was immediately halted upon addition of chloramphenicol and was found to be repressed by glucose and lactate. These results suggest that synthesis of proteolytic extracellular enzymes by the organism studied is controlled by an efficient regulatory mechanism, in which growth rate is an important parameter.     

Formation ofdl-Alanine byCorynebacterium sp. 9366-EMS/270

After a 4-d cultivation in a medium containing 7.5% glucose, 0.6% ammonium nitrate and 0.5% peptone, the leaky mutant Corynebacterium sp. 9366-EMS/270 stimulated in its growth by arginine, was found to accumulate 12.2 g DL-alanine per litre medium.