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At least 325 records · Page 18Linked to original sources

The fate of ribosomes in Escherichia coli cells starved for a carbon source.

The disappearance of ribosomes in Escherichia coli cells starved for a carbon source was studied. We used a series of mutants, some of them lacking in ribonuclease I(RNase I, EC 2.7.7.17), and other containing various combinations of modified polynucleotide phosphorylase (PNPase, EC 2.7.7.8) and modified ribonuclease II (RNase II, EC 3.1.4.1). RNA was prepared from the starved mutant cells and separated on polyacrylamide gels. The results obtained indicate that 23 S RNA degradation is similar in all strains that lack RNase I, and is slightly increased in the strain that contains this enzyme. The extent of 16 S RNA degradation is identical in all strains tested. RNA species in the size of 4 S and smaller accumulate in mutants containing modified forms of PNPase and RNase II. The appearance of an RNA species 10% smaller than 16 S RNA (d16 S RNA) was observed in all strains that contain unmodified RNase II. Analysis of ribosomes and polysomes and their RNA content indicated that polysomes are converted to monosomes and these, in turn, to ribosomal subunits. No RNA degradation products were found in polysomes, 70 S, OR 50 C particle; 30 S subunits contained 16 S RNA as well as the d16 S RNA species. Subunits are degraded to a similar extent in all strains lacking RNase I, and at a slightly faster rate in the strain that contains RNase I. The RNA to protein ratio in subunits prepared from starved cells is similar to that of unstarved cultures. Very little degradation of ribosomal proteins occurs in these mutants during carbon starvation. The proteins released from degraded ribosomes are found in the fast sedimenting (20,000 times g) pellet. Cell viability studies indicated a direct correlation between the capacity of the mutants to recovery from starvation and their capacity to degrade RNA. Thus a biological necessity for degradation of ribosomes during starvation is implied. Based on these data we propose that the endonucleolytic degradation of ribosomal RNA is the primary event in starvation degradation. It takes place in ribosomal subunits, which fall apart after the endonucleoltic attack. The RNA pieces produced by this cleavage are degraded to nucleotide by RNase II and PNPase. The ribosomal proteins attach to the cell membrane.

Bacterial Proteins↗

Carbon source utilization and accumulation of respiration-related substances by freshwater Thioploca species.

Carbon source utilization of Thioploca species from freshwater and brackish lakes in Japan was investigated. Microautoradiography demonstrated that freshwater and brackish Thioploca samples assimilate acetate. In addition, vertical nitrate transportation by freshwater Thioploca was examined by measuring substances accumulated in Thioploca filaments. The filaments of Thioploca sp. from Lake Biwa, a Japanese mesotrophic lake, contained nitrate at concentrations higher than ambient by two to three orders of magnitude. They also accumulated high concentrations of sulfate and abundant elemental sulfur. The results suggest that the Thioploca-specific strategy for sulfur oxidation, migration with accumulated nitrate, is effective even in freshwater habitats of lower sulfide supply.

Ecosystem↗

Mineralization of the herbicide atrazine as a carbon source by a Pseudomonas strain.

Strain YAYA6 was isolated from a mixed microbial community that was growing on atrazine as a sole carbon source and formed quantitative amounts of chloride and nitrate. This strain was identified as a member of the true pseudomonad group (RNA group I) and was given the designation DMS 93-99. The growth yield when atrazine was the sole carbon and nitrogen source was 80 g (dry weight) of cells per mol of atrazine, and the cell doubling time was around 11 h. Approximately 20% of [U-ring 14C]atrazine was mineralized during primary degradation of atrazine. After atrazine disappeared from the culture supernatant, mineralization continued until the level of mineralization was more than 50%. Under different experimental conditions 10% of the atrazine supplied initially was converted to cyanuric acid and < 1% was converted to other s-triazines after prolonged incubation. Degradation proceeded via dechlorination and N-dealkylation. Atrazine was degraded until the concentration was circa 0.1 milligrams/liter. We obtained evidence showing that strain YAYA6 has specific uptake mechanisms for atrazine but less specific degradation mechanisms for s-triazines.

Atrazine↗

On the regulation of adenosine 3', 5'-monophosphate synthesis in bacteria. I. Effect of carbon source variation on cyclic AMP synthesis in Escherichia coli B/r.

1. The effect of carbon source variation in bacterial growth media on their growth rate, inducible enzyme and cyclic AMP synthesis was examined: an inverse relationship between the culture's growth rate and its differential rate of inducible enzyme (tryptophanase and beta-galactosidase), and cyclic AMP synthesis was found. 2. The effect of the culture's growth phase on its sensitivity or resistance to glucose catabolite repression was determined in the wild type and a catabolite insensitive mutant (ABDROI): the wild type's sensitivity to glucose repression was not affected, whereas the insensitivity of the mutant was found to be limited to its early logarithmic phase of growth. At late log, or stationary phase, the mutant was found to be sensitive to glucose repression. 3. Examination of the kinetics of glucose uptake by the mutant, using alpha-[1 4-C] methyl-glucoside showed evidence for two transport systems each with a different affinity to glucose. A low affinity transport system (apparent Km of 3.4-10-minus 5 M) which appears mostly at the early logarithmic phase of growth. A high affinity transport system (apparent Km of 1.2-10-minus 5 M) which appears mostly at the late log and stationary phases of growth. 4. The effect of the culture density variation on its sensitivity to glucose repression showed that sensitivity to glucose catabolic repression is primarily a reflection of the formation of an allosteric effector molecule between glucose and its specific transport molecule which in turn regulates the activity of the adenylate cyclase.

Allosteric Regulation↗

Glutamine assimilation pathways in Neurospora crassa growing on glutamine as sole nitrogen and carbon source.

Neurospora crassa wild-type is almost unable to grow on glutamine as sole nitrogen and carbon source but a GDH-; GS +/- double mutant strain, lacking NADP-dependent glutamate dehydrogenase and partially lacking glutamine synthetase did grow. Under these conditions, the double mutant had a higher chemical energy content than the wild-type. Enzyme assays and labelling experiments with glutamine indicated that in the double mutant glutamine was degraded to ammonium and to carbon skeletons by glutamate synthase, the catabolic (NADH-dependent) glutamate dehydrogenase and the glutamine transaminase-omega-amidase pathway.

Amino Acids↗

Denitrification potential and rates of complex carbon source from dairy effluents in activated sludge system.

A predictive model for the denitrification performance of complex carbon sources was proposed based on compositional data. Potential and rates of denitrification of single dairy components (lactose, lactate, proteins, fat), as well as binary and complex (modelled "process water") mixtures were assessed using test for nitrogen uptake rate (NUR). In all experiments, denitrification potential of mixtures was found to be significantly higher than the sum of individual potentials and denitrification rate with the readily biodegradable moiety of the mixtures was similar to the highest rate obtained with individual components (lactose or lactate). This work shows that activated sludge acclimated to dairy components can be modelled as a single biomass where the maximal anoxic growth rate of the biomass limits the denitrification rate with dairy components. As a consequence, lactose or lactate determine the maximal denitrification rate possible using dairy effluents.

Carbon↗

Glucose-triggered signalling in Saccharomyces cerevisiae: different requirements for sugar phosphorylation between cells grown on glucose and those grown on non-fermentable carbon sources.

Addition of glucose or fructose to cells of the yeast Saccharomyces cerevisiae grown on a nonfermentable carbon source triggers within a few minutes posttranslational activation of trehalase, repression of the CTT1 (catalase) and SSA3 (Hsp70) genes, and induction of the ribosomal protein genes RPL1, RPL25 and RPS33. By using appropriate sugar kinase mutants, it was shown that rapid glucose- or fructose-induced activation of trehalase requires phosphorylation of the sugar. On the other hand, partial induction of RPL1, RPL25 and RPS33 as well as partial repression of CTT1 and SSA3 were observed in the absence of sugar phosphorylation. In glucose-grown nitrogen-starved yeast cells readdition of a nitrogen source triggers activation of trehalase in a glucose- or fructose-dependent way, but with no apparent requirements for phosphorylation of the sugar. Repression of CTT1 and SSA3 under the same conditions was also largely dependent on the presence of the sugar and also in these cases there was a strong effect when the sugar could not be phosphorylated. Nitrogen induction of RPL1, RPL25 and RPS33 was much less dependent on the presence of the sugar, and only phosphorylated sugar caused a further increase in expression. These results show that two glucose-dependent signalling pathways, which can be distinguished on the basis of their requirement for glucose phosphorylation, appear to be involved in activation of trehalase, repression of CTT1 and SSA3 and induction of ribosomal protein genes. They also show that nutrient-induced repression of CTT1 and SSA3 is not a response to improvement of the growth conditions because the addition of nonmetabolizable sugar does not ameliorate the growth conditions. Similarly, the upshift in ribosomal protein synthesis cannot be a response to increased availability of energy or biosynthetic capacity derived from glucose, but it is apparently triggered to a significant extent by specific detection of glucose as such.

Carbohydrate Metabolism↗

Optimizing conditions for poly(beta-hydroxybutyrate) production by Halomonas boliviensis LC1 in batch culture with sucrose as carbon source.

Halomonas boliviensis LC1 is able to accumulate poly(beta-hydroxybutyrate) (PHB) under conditions of excess carbon source and depletion of essential nutrients. This study was aimed at an efficient production of PHB by growing H. boliviensis to high cell concentrations in batch cultures. The effect of ammonium, phosphate, and yeast extract concentrations on cell concentration [cell dry weight (CDW)] and PHB content of H. boliviensis cultured in shake flasks was assayed using a factorial design. High concentrations of these nutrients led to increments in cell growth but reduced the PHB content to some extent. Cultivations of H. boliviensis under controlled conditions in a fermentor using 1.5% (w/v) yeast extract as N source, and intermittent addition of sucrose to provide excess C source, resulted in a polymer accumulation of 44 wt.% and 12 g l(-1) CDW after 24 h of cultivation. Batch cultures in a fermentor with initial concentrations of 2.5% (w/v) sucrose and 1.5% (w/v) yeast extract, and with induced oxygen limitation, resulted in an optimum PHB accumulation, PHB concentration and CDW of 54 wt.%, 7.7 g l(-1) and 14 g l(-1), respectively, after 19 h of cultivation. The addition of casaminoacids in the medium increased the CDW to 14.4 g l(-1) in 17 h but reduced the PHB content in the cells to 52 wt.%.

Culture Techniques↗

Antigenic variation and increase in pathogenicity in Pseudomonas aeruginosa as a result of growth on glucose or N-hexadecane as sole carbon source.

When Pseudomonas aeruginosa is grown on glucose as opposed to n-hexadecane as the sole carbon source, the antigenicity, virulence, and protein composition of the outer membrane are altered. The hydrocarbon-grown cells demonstrate a 3-log increase in virulence over the glucose-grown cells (in mice). There also appears to be an additional protein present in the outer membrane of the n-hexadecane-grown cells. This protein may contribute to the observed antigenic differences between the two cell types.

Animals↗

[Denitrification using PBS as carbon source and biofilm supporter: effect of pH].

PBS, a new kind of biodegradable polymers (BDPs), was used as carbon source and biofilm carrier to remove nitrate from drinking water. The effect of pH was investigated in detail and the results show that fine biofilm can form on the surface of PBS granules and protect denitrifiers. PBS denitrification system can endure higher pH shocking than that of traditional carrier. When influent pH was between 5.0 to approximately 9.0, effluent pH tended to be neutral and denitrification rate was 0.60 to approximately 0.63 mg/(g x d) while the maximum value was 0.70 mg/(g x d) (pH 7.5 to approximately 8.0). Effluent nitrite level was unstable when pH of solution was 6 to approximately 8. It was as high as 0.7 mg/L when pH was below 6 and no more than 0.1 mg/L when pH was higher than 8.

Biodegradation, Environmental↗

Reversible and permanent effects of the carbon sources and various antibiotics on the morphology and metabolic properties of Ustilago cynodontis cells.

The effects of various carbon sources and of antibiotics on the morphology of hypha cells of the fungus Ustilago cynodontis is described. Nonfermentable substrates promote readily reversible yeastlike colonies from hypha cells: all the hypha cells spread on these substrates give rise to yeastlike colonies that revert to the mycelial phenotype when transferred to glucose medium. Among the antibiotics tested, chloramphenicol (CAP) is found to promote, under certain circumstances, a long-lasting, even permanent modification on the morphology of the colonies: the colonies developed on CAP-glucose media are yeastlike, and a percentage of them give rise to colonies whose morphology remains yeastlike even on drug-free media: this effect is also obtained with cells cultivated in liquid medium. This permanent morphological modification is accompanied by a change of metabolic properties. Similar permanent effects are obtained with ethidium bromide, suggesting that mitochondrial functioning is involved in these modifications.

Anti-Bacterial Agents↗

Studies on a novel carbon source and cosolvent for lipase production by Candida rugosa.

Oleic acid esters were shown to be the best carbon source for both cell growth and lipase production by Candida rugosa. Use of a cosolvent, dodecane, in fermentations improved the solubility of solid substrates and increased oxygen solubility. This resulted in the highest lipase activity in batch fermentation with glycerol trioleate and dodecane. Lipase activity reached 77.1 units ml(-1).

Alkanes↗

Strain-dependent variation in carbon source regulation of nucleus-encoded mitochondrial proteins of Saccharomyces cerevisiae.

Nuclear genes encoding mitochondrial proteins are regulated by carbon source with significant heterogeneity among four Saccharomyces cerevisiae strains. This strain-dependent variation is seen both in respiratory capacity of the cells and in the expression of beta-galactosidase reporter fusions to the promoters of CYB2, CYC1, CYC3, MnSOD, and RPO41.

Bacterial Proteins↗

Xylose and some non-sugar carbon sources cause catabolite repression in Saccharomyces cerevisiae.

Glucose and other sugars, such as galactose or maltose, are able to cause carbon catabolite repression in Saccharomyces cerevisiae. Although glycolytic intermediates have been suggested as signal for repression, no evidence for such a control mechanism is available. The establishment of a correlation between levels of intracellular metabolites and the extent of catabolite repression may facilitate the identification of potential signal molecules in the process. To set a framework for such a study, the repression produced by xylose, glycerol and dihydroxyacetone upon genes belonging to different repressible circuits was tested, using an engineered strain of S. cerevisiae able to metabolize xylose. Xylose decreased the derepression of various enzymes in the presence of ethanol by at least 10-fold; the corresponding mRNAs were not detected in these conditions. Xylose also impaired the derepression of galactokinase and invertase. Glycerol and dihydroxyacetone decreased 2- to 3-fold the derepression observed in ethanol or galactose but did not affect invertase derepression. For yeast cells grown in media with different carbon sources, no correlation was found between repression of fructose-1,6-bisphosphatase and intracellular levels of glucose 6-phosphate or fructose 1,6-bisphosphate.

Carbon↗

Carbon sources and trophic position of the main species of fishes of Baía River, Paraná River floodplain, Brazil.

In order to verify the carbon source and trophic position of the main species of fishes, of the Paraná River floodplain, we analysed the proportion of stable carbon (delta 13C) and nitrogen (delta 15N) isotopes in muscle of fishes sampled in the rainy season. We analyzed adult individuals of Loricariichthys platymetopon, Schizodon borellii, Leporinus lacustris, Auchenipterus osteomystax, Iheringichthys labrosus, Leporinus friderici, and Serrasalmus marginatus. These data were compared with the results obtained by the analyzing stomach contents. The primary producers found in the Baía River were the C3 plants (riparian vegetation, macrophytes, periphyton, and phytoplankton) and the C4 plants (macrophytes). The results of the contribution analysis revealed that the carbon used by the species was derived from C3 plants. According to the trophic position estimates (diet and delta 15N), the species primarily consumed Loricariichthys platymetopon, Schizodon borellii, Leporinus lacustris, and Leporinus friderici and, secondarily Auchenipterus osteomystax, Iheringichthys labrosus, and Serrasalmus marginatus. There was no significant difference between the two methods utilized.

Animals↗

Analysis of factors affecting the accuracy, reproducibility, and interpretation of microbial community carbon source utilization patterns.

We determined factors that affect responses of bacterial isolates and model bacterial communities to the 95 carbon substrates in Biolog microtiter plates. For isolates and communities of three to six bacterial strains, substrate oxidation rates were typically nonlinear and were delayed by dilution of the inoculum. When inoculum density was controlled, patterns of positive and negative responses exhibited by microbial communities to each of the carbon sources were reproducible. Rates and extents of substrate oxidation by the communities were also reproducible but were not simply the sum of those exhibited by community members when tested separately. Replicates of the same model community clustered when analyzed by principal-components analysis (PCA), and model communities with different compositions were clearly separated on the first PCA axis, which accounted for >60% of the dataset variation. PCA discrimination among different model communities depended on the extent to which specific substrates were oxidized. However, the substrates interpreted by PCA to be most significant in distinguishing the communities changed with reading time, reflecting the nonlinearity of substrate oxidation rates. Although whole-community substrate utilization profiles were reproducible signatures for a given community, the extent of oxidation of specific substrates and the numbers or activities of microorganisms using those substrates in a given community were not correlated. Replicate soil samples varied significantly in the rate and extent of oxidation of seven tested substrates, suggesting microscale heterogeneity in composition of the soil microbial community.

Journal Article↗

Growth, enzyme levels, and some metabolic properties of an Escherichia coli mutant grown on L-threonine as the sole carbon source.

A mutant of Escherichia coli (designated E. coli SBD-76) that utilizes L-threonine as the sole carbon source was isolated. In contrast with levels in extracts of wild-type cells, the levels of threonine dehydrogenase in extracts of this mutant were 100-fold higher than levels of threonine aldolase or degradative threonine dehydratase. Catabolite repression of threonine dehydrogenase was manifested in wild-type, but not SBD-76, cells. For purposes of isolating enzymes, large quantities of SBD-76 cells with the elevated threonine dehydrogenase level could be grown in a fermentor in modified Fraser medium containing 1% glycerol, rather than in the 0.2% L-threonine minimal medium used to isolate the mutant. SBD-76 cells grown on L-threonine excreted glycine and aminoacetone into the medium, and extracts of the mutant strain catalyzed a quantitative conversion of L-threonine to glycine and aminoacetone.

Acetone↗