scholarly journals Aspartate Biosynthesis Is Essential for the Growth of Streptococcus thermophilus in Milk, and Aspartate Availability Modulates the Level of Urease Activity

2007 ◽  
Vol 73 (18) ◽  
pp. 5789-5796 ◽  
Author(s):  
Stefania Arioli ◽  
Christophe Monnet ◽  
Simone Guglielmetti ◽  
Carlo Parini ◽  
Ivano De Noni ◽  
...  
Keyword(s):  
Aspartic Acid ◽  
Phosphoenolpyruvate Carboxylase ◽  
Parent Strain ◽  
Urease Activity ◽  
Streptococcus Thermophilus ◽  
Defined Medium ◽  
Lactobacillus Delbrueckii ◽  
Chemically Defined Medium ◽  
Ammonium Ions ◽  
Carboxylase Activity

ABSTRACT We investigated the carbon dioxide metabolism of Streptococcus thermophilus, evaluating the phenotype of a phosphoenolpyruvate carboxylase-negative mutant obtained by replacement of a functional ppc gene with a deleted and inactive version, Δppc. The growth of the mutant was compared to that of the parent strain in a chemically defined medium and in milk, supplemented or not with l-aspartic acid, the final product of the metabolic pathway governed by phosphoenolpyruvate carboxylase. It was concluded that aspartate present in milk is not sufficient for the growth of S. thermophilus. As a consequence, phosphoenolpyruvate carboxylase activity was considered fundamental for the biosynthesis of l-aspartic acid in S. thermophilus metabolism. This enzymatic activity is therefore essential for growth of S. thermophilus in milk even if S. thermophilus was cultured in association with proteinase-positive Lactobacillus delbrueckii subsp. bulgaricus. It was furthermore observed that the supplementation of milk with aspartate significantly affected the level of urease activity. Further experiments, carried out with a p ureI -gusA recombinant strain, revealed that expression of the urease operon was sensitive to the aspartate concentration in milk and to the cell availability of glutamate, glutamine, and ammonium ions.

scholarly journals Physiological Study of Lactobacillus delbrueckii subsp. bulgaricus Strains in a Novel Chemically Defined Medium

2000 ◽  
Vol 66 (12) ◽  
pp. 5306-5311 ◽  
Author(s):  
Christian Chervaux ◽  
S. Dusko Ehrlich ◽  
Emmanuelle Maguin
Keyword(s):  
Carbon Sources ◽  
Streptococcus Thermophilus ◽  
Defined Medium ◽  
Lactobacillus Delbrueckii ◽  
Protein Labeling ◽  
Chemically Defined Medium ◽  
Phosphotransferase System ◽  
Nutritional Conditions ◽  
Labeling Method

ABSTRACT We developed a chemically defined medium called milieu proche du lait (MPL), in which 22 Lactobacillus delbrueckii subsp.bulgaricus (L. bulgaricus) strains exhibited growth rates ranging from 0.55 to 1 h−1. MPL can also be used for cultivation of other lactobacilli and Streptococcus thermophilus. The growth characteristics of L. bulgaricus in MPL containing different carbon sources were determined, including an initial characterization of the phosphotransferase system transporters involved. For the 22 tested strains, growth on lactose was faster than on glucose, mannose, and fructose. Lactose concentrations below 0.4% were limiting for growth. We isolated 2-deoxyglucose-resistant mutants from strains CNRZ397 and ATCC 11842. CNRZ397-derived mutants were all deficient for glucose, fructose, and mannose utilization, indicating that these three sugars are probably transported via a unique mannose-specific-enzyme-II-like transporter. In contrast, mutants of ATCC 11842 exhibited diverse phenotypes, suggesting that multiple transporters may exist in that strain. We also developed a protein labeling method and verified that exopolysaccharide production and phage infection can occur in MPL. The MPL medium should thus be useful in conducting physiological studies ofL. bulgaricus and other lactic acid bacteria under well controlled nutritional conditions.

Mise en évidence d'une activité uréasique chez Streptococcus thermophilus

1988 ◽  
Vol 34 (6) ◽  
pp. 818-822 ◽  
Author(s):  
V. Juillard ◽  
M. J. Desmazeaud ◽  
H. E. Spinnler
Keyword(s):  
Amino Acids ◽  
Urease Activity ◽  
Culture Medium ◽  
Colorimetric Method ◽  
Streptococcus Thermophilus ◽  
Stationary Growth Phase ◽  
Optimal Temperature ◽  
Ammonium Ions ◽  
Stationary Growth ◽  
Optimal Ph

In Streptococcus thermophilus CNRZ 404, the presence of urease activity was demonstrated by means of a specific colorimetric method for ammonium ions. The main physicochemical properties of the enzyme were determined. The Km with urea as substrate was 1.19 mM and the optimal pH was approximately 7.5. Because both thermolability and enzyme activity increased as the temperature was increased to 70 °C, the optimal temperature could not be determined with precision. Urease activity was maximal at the beginning of the stationary growth phase; it was stimulated by the presence of urea and of certain amino acids such as arginine and glutamic acid in the culture medium. This activity has been detected in several other strains of Streptococcus thermophilus. [Translated by the journal]

The specificity of oligopeptide transport by Streptococcus thermophilus resembles that of Lactococcus lactis and not that of pathogenic streptococci

Microbiology ◽  
2005 ◽  
Vol 151 (6) ◽  
pp. 1987-1994 ◽  
Author(s):  
Odile Juille ◽  
Dominique Le Bars ◽  
Vincent Juillard
Keyword(s):  
Mass Spectrometry ◽  
Amino Acids ◽  
Time Course ◽  
Streptococcus Thermophilus ◽  
Defined Medium ◽  
Peptide Transport ◽  
Chemically Defined Medium ◽  
Crucial Step ◽  
Tryptic Digest ◽  
Transport Studies

Peptide transport is a crucial step in the growth of Streptococcus thermophilus in protein- or peptide-containing media. The objective of the present work was to determine the specificity of peptide utilization by this widely used lactic acid bacterium. To reach that goal, complementary approaches were employed. The capability of a proteinase-negative S. thermophilus strain to grow in a chemically defined medium containing a mixture of peptides isolated from milk as the source of amino acids was analysed. Peptides were separated into three size classes by ultrafiltration. The strain was able to use peptides up to 3·5 kDa during growth, as revealed by liquid chromatography and mass spectrometry analyses. The same strain was grown in chemically defined medium containing a tryptic digest of casein, and the respective time-course consumption of the peptides during growth was estimated. The ability to consume large peptides (up to 23 residues) was confirmed, as long as they are cationic and hydrophobic. These results were confirmed by peptide transport studies. Extension of the study to 11 other strains revealed that they all shared these preferences.

Nitrogen source regulates expression of alanine dehydrogenase isoenzymes in Streptomyces avermitilis in a chemically defined medium

1997 ◽  
Vol 43 (2) ◽  
pp. 189-193 ◽  
Author(s):  
Jan Novák ◽  
Jan Kopecký ◽  
Zdenko Vaněk
Keyword(s):  
Nitrogen Source ◽  
Ammonium Sulfate ◽  
Alanine Aminotransferase ◽  
Specific Activity ◽  
Defined Medium ◽  
Streptomyces Avermitilis ◽  
Chemically Defined Medium ◽  
Ammonium Ions ◽  
Alanine Dehydrogenase ◽  
Dehydrogenase Reaction

Ammonium ions and alanine influence production of the macrolide avermectin in Streptomyces avermitilis. L-Alanine dehydrogenase and alanine aminotransferase are the primary enzymes responsible for regulating the intracellular concentration of alanine and also of ammonium ions. In cultures of S. avermitilis in a chemically defined medium with ammonia or L-alanine as the only nitrogen source, specific activities of both enzymes increased during growth. The alanine dehydrogenase specific activity increased more than 86-fold after the culture was supplemented with 0.2% L-alanine and 5-fold after addition of 0.5% ammonium sulfate, whereas alanine aminotransferase specific activity increased 3- to 4-fold with either substrate. Five isoenzymes of alanine dehydrogenase were detected histochemically in S. avermitilis after native gel electrophoresis. Isoenzyme 1 was induced by alanine and temporarily repressed by high concentrations of ammonium sulfate. The presence of isoenzyme 1 was also related to changes in the kinetic properties of the alanine dehydrogenase reaction measured in crude desalted extracts. A nonlinear double-reciprocal plot was obtained in initial velocity studies using L-alanine as a substrate in the sample induced with L-alanine. The nonlinearity was caused by both substrate inhibition and allosteric regulation (positive cooperativity) by L-alanine. In contrast, the sample induced by ammonium sulfate showed a linear double-reciprocal plot.Key words: isoenzymes, L-alanine dehydrogenase, Streptomyces avermitilis, avermectin.

A Deficiency in Aspartate Biosynthesis inLactococcus lactis subsp. lactis C2 Causes Slow Milk Coagulation

1998 ◽  
Vol 64 (5) ◽  
pp. 1673-1679 ◽  
Author(s):  
Hua Wang ◽  
Weizhu Yu ◽  
Tim Coolbear ◽  
Dan O’Sullivan ◽  
Larry L. McKay
Keyword(s):  
Carbon Dioxide ◽  
Amino Acid ◽  
Aspartic Acid ◽  
Pyruvate Carboxylase ◽  
Defined Medium ◽  
Transport Systems ◽  
Chemically Defined Medium ◽  
Acid Transport ◽  
Amino Acid Requirements ◽  
Milk Coagulation

ABSTRACT A mutant of fast milk-coagulating (Fmc+)Lactococcus lactis subsp. lactis C2, designatedL. lactis KB4, was identified. Although possessing the known components essential for utilizing casein as a nitrogen source, which include functional proteinase (PrtP) activity and oligopeptide, di- and tripeptide, and amino acid transport systems, KB4 exhibited a slow milk coagulation (Fmc−) phenotype. When the amino acid requirements of L. lactis C2 were compared with those of KB4 by use of a chemically defined medium, it was found that KB4 was unable to grow in the absence of aspartic acid. This aspartic acid requirement could also be met by aspartate-containing peptides. The addition of aspartic acid to milk restored the Fmc+phenotype of KB4. KB4 was found to be defective in pyruvate carboxylase and thus was deficient in the ability to form oxaloacetate and hence aspartic acid from pyruvate and carbon dioxide. The results suggest that when lactococci are propagated in milk, aspartate derived from casein is unable to meet fully the nutritional demands of the lactococci, and they become dependent upon aspartate biosynthesis.

Influence of ions on growth and production of exopolysaccharides by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772

2000 ◽  
Vol 67 (1) ◽  
pp. 131-135 ◽  
Author(s):  
GERT J. GROBBEN ◽  
INGEBORG C. BOELS ◽  
JAN SIKKEMA ◽  
MARK R. SMITH ◽  
JAN A. M. DE BONT
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Streptococcus Thermophilus ◽  
Defined Medium ◽  
Lactobacillus Delbrueckii ◽  
Sugar Composition ◽  
Carbohydrate Source ◽  
Medium Components ◽  
Number Of Factors ◽  
Initial Ph

Several lactic acid bacteria produce exopolysaccharides (EPS), either attached to the cell wall or excreted into the environment as slime material. EPS produced by Lactobacillus delbrueckii subsp. bulgaricus (Lb. bulgaricus) and Streptococcus thermophilus play an important role in improving the texture and stability of yogurt and preventing syneresis (Cerning, 1990; Nakajima et al. 1990). The amount and composition of the EPS produced by lactic acid bacteria are dependent on a number of factors, such as temperature, initial pH, carbon source and the availability of minerals, vitamins and other medium components.In previous work it was shown that the production and sugar composition of the EPS from Lb. bulgaricus NCFB2772 are affected by the carbohydrate source (Grobben et al. 1995, 1996). In a simplified defined medium, from which several vitamins and trace elements were omitted, EPS production by Lb. bulgaricus significantly increased, although growth of the strain was reduced (Grobben et al. 1998).

scholarly journals Enhancement of Exopolysaccharide Production byLactobacillus delbrueckii subsp. bulgaricus NCFB 2772 with a Simplified Defined Medium

1998 ◽  
Vol 64 (4) ◽  
pp. 1333-1337 ◽  
Author(s):  
G. J. Grobben ◽  
I. Chin-Joe ◽  
V. A. Kitzen ◽  
I. C. Boels ◽  
F. Boer ◽  
...  
Keyword(s):  
Amino Acids ◽  
Defined Medium ◽  
Complete Loss ◽  
Lactobacillus Delbrueckii ◽  
Complete Medium ◽  
Calcium Pantothenate ◽  
Chemically Defined Medium ◽  
Batch Cultures ◽  
Exopolysaccharide Production ◽  
Medium Components

ABSTRACT The aim of this work was to investigate the medium requirements for growth and production of exopolysaccharides by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772. The strain was grown in batch cultures on a chemically defined medium, and the technique of single omission of medium components was applied to determine the nutritional requirements. The omission of aspartic acid, glutamic acid, or glycine affected growth only slightly, and the omission of glutamine, asparagine, or threonine resulted in a stronger reduction of the growth. All the other amino acids were essential. Multiple omissions of amino acids caused an almost complete loss of growth. L. delbrueckii subsp. bulgaricusrequired only riboflavin, calcium pantothenate, and nicotinic acid as individual vitamins. Surprisingly, when only these vitamins were present in the medium and other vitamins were not, less growth was observed than in the complete medium but the amount of exopolysaccharide produced was significantly greater. These observations were studied in more detail with a simplified defined medium in which L. delbrueckii subsp.bulgaricus was able to grow and produce exopolysaccharides. Although the final optical density in the simplified medium was lower, the production of exopolysaccharides was about twofold higher than in the complete medium.

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.

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