scholarly journals Influence of Glutamate on Growth, Sporulation, and Spore Properties of Bacillus cereus ATCC 14579 in Defined Medium

2005 ◽  
Vol 71 (6) ◽  
pp. 3248-3254 ◽  
Author(s):  
Ynte P. de Vries ◽  
Ratna D. Atmadja ◽  
Luc M. Hornstra ◽  
Willem M. de Vos ◽  
Tjakko Abee
Keyword(s):  
Amino Acids ◽  
Bacillus Cereus ◽  
Exponential Growth ◽  
Dipicolinic Acid ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Defined Medium ◽  
Mother Cells ◽  
Almost All ◽  
Time Frames

ABSTRACT A chemically defined medium in combination with an airlift fermentor system was used to study the growth and sporulation of Bacillus cereus ATCC 14579. The medium contained six amino acids and lactate as the main carbon sources. The amino acids were depleted during exponential growth, while lactate was metabolized mainly during stationary phase. Two concentrations of glutamate were used: high (20 mM; YLHG) and low (2.5 mM; YLLG). Under both conditions, sporulation was complete and synchronous. Sporulation started and was completed while significant amounts of carbon and nitrogen sources were still present in the medium, indicating that starvation was not the trigger for sporulation. Analysis of amino acids and NH4 + in the culture supernatant showed that most of the nitrogen assimilated by the bacteria was taken up during sporulation. The consumption of glutamate depended on the initial concentration; in YLLG, all of the glutamate was used early during exponential growth, while in YLHG, almost all of the glutamate was used during sporulation. In YLLG, but not in YLHG, NH4 + was taken up by the cells during sporulation. The total amount of nitrogen used by the bacteria in YLLG was less than that used by the bacteria in YLHG, although a significant amount of NH4 + was present in the medium throughout sporulation. Despite these differences, growth and temporal expression of key sigma factors involved in sporulation were parallel, indicating that the genetic time frames of sporulation were similar under both conditions. Nevertheless, in YLHG, dipicolinic acid production started later and the spores were released from the mother cells much later than in YLLG. Notably, spores had a higher heat resistance when obtained after growth in YLHG than when obtained after growth in YLLG, and the spores germinated more rapidly and completely in response to inosine, l-alanine, and a combination of these two germinants.

Amino acid uptake systems in Bacteroides ruminicola

1979 ◽  
Vol 25 (10) ◽  
pp. 1161-1168 ◽  
Author(s):  
Roselynn M. W. Stevenson
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Nitrogen Sources ◽  
Amino Acid Uptake ◽  
Defined Medium ◽  
High Rate ◽  
Transport Systems ◽  
Acid Uptake ◽  
Chemical Structures ◽  
Kinetic Inhibition

Uptake of amino acids by Bacteroides ruminicola was observed in cells grown in a complete defined medium, containing ammonia as the nitrogen source. A high rate of uptake occurred only in fresh medium, as an inhibitory substance, possibly acetate, apparently accumulated during growth. All amino acids except proline were taken up and incorporated into cold trichloroacetic acid precipitable material. Different patterns of incorporation and different responses to 2,4-dinitrophenol and potassium ferricyanide indicated multiple uptake systems were involved. Kinetic inhibition patterns suggested six distinct systems were present for amino acid uptake, with specificities related to the chemical structures of the amino acids. Thus, the failure of free amino acids to act as sole nitrogen sources for growth of B. ruminicola is not due to the absence of transport systems for these compounds.

THE IN VITRO RELEASE OF AMINO ACIDS BY RUMEN PAPILLAE

1980 ◽  
Vol 60 (2) ◽  
pp. 281-291 ◽  
Author(s):  
R. J. BOILA ◽  
L. P. MILLIGAN
Keyword(s):  
Amino Acids ◽  
Carbon Sources ◽  
In Vitro Release ◽  
Nitrogen Sources ◽  
Chromatographic Technique ◽  
Salts Of Organic Acids ◽  
Liquid Chromatographic ◽  
Vitro Release ◽  
Effective Source

Rumen papillae from cattle were incubated aerobically with combinations of NH4Cl, amino acids and salts of organic acids, the latter including propionate, pyruvate, α-ketoglutarate and glyoxylate. Amino acids in the incubation media were analyzed using a gas-liquid chromatographic technique entailing separation of the isobutyl-N(0)-heptafluorobutyryl esters: glutamine was recovered with glutamate, asparagine with aspartate, and citrulline with ornithine. Rumen papillae incubated with pyruvate or propionate released alanine, but with the latter substrate only glutamate was effective as a nitrogen source. Glycine and glutamate plus glutamine were released in the presence of glyoxylate and α-ketoglutarate, respectively. Serine and aspartate plus asparagine were not quantitatively major products released by rumen papillae. Glutamate was an effective source of nitrogen for the release of alanine and glycine with pyruvate and glyoxylate, respectively, as carbon sources. When rumen papillae were incubated with pyruvate or glyoxylate as the added carbon source, glutamine nitrogen disappeared and was not accounted for by the amino acids measured. With arginine as a substrate, there was a release of ornithine by rumen papillae indicating urea production. The tissues of rumen papillae appear to synthesize amino acids from expected carbon sources with ammonia or glutamate as nitrogen sources and to catabolize glutamine and arginine. The metabolism of amino acids by rumen papillae would contribute to the interchange of nitrogen between the rumen and the host.

Nitrogen nutrition and regulation of cephalosporin production in Streptomyces clavuligerus

1979 ◽  
Vol 25 (1) ◽  
pp. 61-67 ◽  
Author(s):  
Yair Aharonowitz ◽  
Arnold L. Demain
Keyword(s):  
Amino Acids ◽  
Growth Rate ◽  
Nitrogen Nutrition ◽  
Antibiotic Production ◽  
Inorganic Nitrogen ◽  
Amino Nitrogen ◽  
Nitrogen Sources ◽  
Streptomyces Clavuligerus ◽  
Defined Medium ◽  
Repressive Effect

When used as sole nitrogen source, certain amino acids (e.g., proline, asparagine) supported both growth and sporulation by Streptomyces clavuligerus streaked onto solid defined medium. Ammonium supported growth but suppressed sporulation. Amino nitrogen was best for cephalosporin production in liquid defined medium, although urea was almost as useful. A comparison of amino acids showed asparagine and glutamine to be the best nitrogen sources and arginine to be almost as good. Ammonium salts supported a somewhat lower growth rate than asparagine, but antibiotic production was very poor on these inorganic nitrogen sources. Addition of ammonium to asparagine did not affect growth rate but increased mycelial mass; cephalosporin production was reduced by about 75%. Antibiotic production was more closely associated with growth in the absence of ammonium than in its presence, indicating a strong inhibitory and (or) repressive effect of NH4+ on antibiotic production. Ammonium exerted its negative effect when added at 24 h or earlier, i.e. before antibiotic formation began.

scholarly journals The Growth Of Rhizobium in Synthetic Media

1961 ◽  
Vol 14 (3) ◽  
pp. 349 ◽  
Author(s):  
FJ Bergersen
Keyword(s):  
Amino Acids ◽  
Nitrogen Source ◽  
Exponential Growth ◽  
Defined Medium ◽  
Complete Medium ◽  
Good Growth ◽  
Chemically Defined Medium ◽  
Sodium Glutamate ◽  
Synthetic Media

A chemically defined medium for the growth of Rhizobium is described in which populations of up to 5 x 109 cells/ml were obtained. For the six strains of bacteria studied the complete medium supported exponential growth for two to five generations. The concentrations of biotin giving best growth varied ith strain between 125 and 250 f'g/l when the nitrogen source was sodium glutamate. NHt, NOs, and other amino acids, singly or in combination, did not upport as good growth as did sodium glutamate.

scholarly journals Control of the production of exo-β-N-acetylglucosaminidase by Bacillus subtilis B. Derepression during gluconeogenesis and initial stages of sporulation

1973 ◽  
Vol 134 (1) ◽  
pp. 271-281 ◽  
Author(s):  
Stephen J. Brewer ◽  
Roger C. W. Berkeley
Keyword(s):  
Bacillus Subtilis ◽  
Exponential Growth ◽  
Phosphoenolpyruvate Carboxylase ◽  
Metal Ion ◽  
Tricarboxylic Acid Cycle ◽  
Carbon Sources ◽  
Defined Medium ◽  
Tricarboxylic Acid ◽  
Carboxylase Activity ◽  
Acid Cycle

1. The control of exo-β-N-acetylglucosaminidase (EC 3.2.1.30) production by Bacillus subtilis B growing on a chemically defined medium was studied. 2. The enzyme was repressed during exponential growth by those carbon sources that enter the glycolytic pathway above the level of phosphoenolpyruvate. When exponential growth ceased as a result of low concentrations of the nitrogen, carbon or metal ion components of the medium, the enzyme was formed and its amount could be increased by the addition of cell-wall fragments as inducer. 3. The enzyme was de-repressed and could be induced during exponential growth on non-glycolytic compounds metabolized directly into pyruvate, acetyl-CoA or tricarboxylic acid cycle intermediates. 4. The major difference in the metabolism of the organism utilizing these two groups of compound was the existence of high activities of phosphoenolpyruvate carboxylase required for gluconeogenesis. 5. It is concluded that the de-repression of glucosaminidase occurs when the only principal change detected in the intermediary metabolism of the organism was the presence of high activities of phosphoenolpyruvate carboxylase. 6. When the organism was grown on media containing repressing compounds, the enzyme was only de-repressed on entry of the cells into the initial stages of sporulation, where phosphoenolpyruvate carboxylase activity, even in the presence of excess of glucose, increased in parallel with glucosaminidase, neutral proteinase and alkaline phosphatase activities. 7. These results suggest a strong link, at the level of the tricarboxylic acid cycle, between the control of phosphoenolpyruvate carboxylase and the control of the de-repression of glucosaminidase and sporulation.

scholarly journals Influence of Amino Acid Feeding on Production of Calcimycin and Analogs in Streptomyces chartreusis

2021 ◽  
Vol 18 (16) ◽  
pp. 8740
Author(s):  
Kirstin I. Arend ◽  
Julia E. Bandow
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Nitrogen Sources ◽  
Gene Clusters ◽  
Biosynthetic Gene Cluster ◽  
Defined Medium ◽  
Glutamine Supplementation ◽  
Biosynthetic Gene ◽  
Biosynthetic Gene Clusters

Streptomyces chartreusis NRRL 3882 produces the polyether ionophore calcimycin and a variety of analogs, which originate from the same biosynthetic gene cluster. The role of calcimycin and its analogs for the producer is unknown, but calcimycin has strong antibacterial activity. Feeding experiments were performed in chemically defined medium systematically supplemented with proteinogenic amino acids to analyze their individual effects on calcimycin synthesis. In the culture supernatants, in addition to known calcimycin analogs, eight so far unknown analogs were detected using LC-MS/MS. Under most conditions cezomycin was the compound produced in highest amounts. The highest production of calcimycin was detected upon feeding with glutamine. Supplementation of the medium with glutamic acid resulted in a decrease in calcimycin production, and supplementation of other amino acids such as tryptophan, lysine, and valine resulted in the decrease in the synthesis of calcimycin and of the known intermediates of the biosynthetic pathway. We demonstrated that the production of calcimycin and its analogs is strongly dependent on amino acid supply. Utilization of amino acids as precursors and as nitrogen sources seem to critically influence calcimycin synthesis. Even amino acids not serving as direct precursors resulted in a different product profile regarding the stoichiometry of calcimycin analogs. Only slight changes in cultivation conditions can lead to major changes in the metabolic output, which highlights the hidden potential of biosynthetic gene clusters. We emphasize the need to further study the extent of this potential to understand the ecological role of metabolite diversity originating from single biosynthetic gene clusters.

scholarly journals Role of Nitrogen and Carbon Transport, Regulation, and Metabolism Genes for Saccharomyces cerevisiae Survival In Vivo

2006 ◽  
Vol 5 (5) ◽  
pp. 816-824 ◽  
Author(s):  
Joanne M. Kingsbury ◽  
Alan L. Goldstein ◽  
John H. McCusker
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acids ◽  
Drug Targets ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Opportunistic Pathogen ◽  
Close Relative ◽  
Environment Interaction ◽  
Nitrogenous Compounds

ABSTRACT Saccharomyces cerevisiae is both an emerging opportunistic pathogen and a close relative of pathogenic Candida species. To better understand the ecology of fungal infection, we investigated the importance of pathways involved in uptake, metabolism, and biosynthesis of nitrogen and carbon compounds for survival of a clinical S. cerevisiae strain in a murine host. Potential nitrogen sources in vivo include ammonium, urea, and amino acids, while potential carbon sources include glucose, lactate, pyruvate, and fatty acids. Using mutants unable to either transport or utilize these compounds, we demonstrated that no individual nitrogen source was essential, while glucose was the most significant primary carbon source for yeast survival in vivo. Hydrolysis of the storage carbohydrate glycogen made a slight contribution for in vivo survival compared with a substantial requirement for trehalose hydrolysis. The ability to sense and respond to low glucose concentrations was also important for survival. In contrast, there was little or no requirement in vivo in this assay for any of the nitrogen-sensing pathways, nitrogen catabolite repression, the ammonium- or amino acid-sensing pathways, or general control. By using auxotrophic mutants, we found that some nitrogenous compounds (polyamines, methionine, and lysine) can be acquired from the host, while others (threonine, aromatic amino acids, isoleucine, and valine) must be synthesized by the pathogen. Our studies provide insights into the yeast-host environment interaction and identify potential antifungal drug targets.

THE METABOLISM OF YEAST SPORULATION: V. STIMULATION AND INHIBITION OF SPORULATION AND GROWTH BY NITROGEN COMPOUNDS

1963 ◽  
Vol 9 (2) ◽  
pp. 259-277 ◽  
Author(s):  
J. J. Miller
Keyword(s):  
Amino Acids ◽  
Glutamic Acid ◽  
Ammonium Sulphate ◽  
Slow Growth ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Nitrogen Compounds ◽  
Sporulation Medium ◽  
Carbon And Nitrogen ◽  
Low Concentrations

The effects of a variety of nitrogen compounds, mainly amino acids, on ascus production by S. cerevisiae in sporulation media containing several carbon sources were compared. With acetate, glucose, ethanol, pyruvate, and lactate all the amino acids (0.01 M) and most other nitrogen compounds inhibited sporulation. With dihydroxyacetone fewer nitrogen compounds inhibited and alanine, phenylalanine, glycine, and lysine (especially the latter) increased sporulation, even at low concentrations (0.0001 M). Sporulation in buffer resembled sporulation in dihydroxyacetone in its response to amino acids. For greatest inhibition of sporulation the simultaneous presence of carbon and nitrogen sources was necessary. With dihydroxyacetone as carbon source, glutamic acid increased the average number of spores per ascus, but decreased it with all other carbon sources.No correlation was evident between the effects of the amino acids on sporulation and their value as nitrogen sources for growth or with changes in the oxygen uptake of cells in their presence. Dihydroxyacetone did not support growth when the nitrogen source was ammonium sulphate, but slow growth resulted when ammonium sulphate was replaced by glutamic acid or casein hydrolyzate.A sporulation medium is recommended containing, in water (or buffer) agar, 0.02 M dihydroxyacetone and 0.01 M lysine.

Regulation of nitrate reductase by various nitrogen sources in cultured Ipomoea sp.

1982 ◽  
Vol 60 (4) ◽  
pp. 386-396 ◽  
Author(s):  
M. W. Zink
Keyword(s):  
Amino Acids ◽  
Nitrate Reductase ◽  
Reductase Activity ◽  
Casein Hydrolysate ◽  
Nitrogen Sources ◽  
Defined Medium ◽  
Morning Glory ◽  
Stages Of Growth ◽  
Activity Peak ◽  
Nitrogen Phosphorus

The nitrate reductase of Ipomoea sp. (morning glory) grown in suspension culture on a chemically defined medium is induced by nitrate. The developmental pattern of the enzyme shows three peaks of enzyme activity: peak I during the initial stages of growth, peak II during the exponential growth, and peak III just before the onset of the stationary phase of growth. The full induction of nitrate reductase by nitrate is dependent on the availability of a source of energy, nitrogen, phosphorus, and sulfur. After 24 h of growth high levels of nitrate reductase are detected in the presence of casein hydrolysate and ammonium pyrrolidine carboxylate and an appreciable level in the presence of nitrite and ammonium as nitrogen sources. Glutamine, asparagine, and amino acids are also regulators of nitrate reductase synthesis whether they are used as sole nitrogen sources or as additives in nitrate media. When used as sole nitrogen source the amino acids generally fall into three distinct classes. Amino acids in the first class promote an increase in the level of nitrate reductase during the initial stages of growth. Those in the second class promote a rapid decay of the enzyme while those in the third class initially cause enzyme decay but then promote the synthesis of the enzyme on further growth. Interactions between amino acids as nitrogen sources have been found to occur. The possible reasons for the three peaks of nitrate reductase activity when Ipomoea is grown on nitrate are discussed.

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