[Ecology of fermentation sarcinas Sarcina ventriculi and Sarcina maxima].
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Animals↗
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Phylogenetic placement of Sarcina ventriculi and Sarcina maxima within group I Clostridium, a possible problem for future revision of the genus Clostridium. Request for an opinion.
The 16S rRNA gene sequences of Sarcina ventriculi DSM 286T (T = type strain) and Sarcina maxima DSM 316T were determined. Phylogenetic analysis revealed that these two species are closely related to each other and belong to group I Clostridium (sensu Johnson and Francis). The implications of these phylogenetic findings for future revision of the genus Clostridium are discussed.
Clostridium↗
Fine structure of Sarcina maxima and Sarcina ventriculi.
The fine structure of Sarcina maxima and S. ventriculi was studied by electron and phase-contrast microscopy. The two organisms differ mainly with respect to their cell surface. A thick cellulose layer present on the cell wall of S. ventriculi was not observed on the surface of S. maxima. Carbon replication indicated that the outer surface of S. ventriculi is rough in contour, probably as the result of the fibrillar nature of the accumulated cellulose. The cytoplasm of both sarcinae contains inclusions similar to polysaccharide and polymetaphosphate granules. Mesosomes were observed in cells of S. maxima. Packets of S. ventriculi generally comprise a larger number of cells and are more irregularly constructed than those of S. maxima. Cells in large packets of S. ventriculi assume flattened or otherwise irregular shapes, whereas cells of S. maxima maintain a more uniform appearance.
Microscopy, Electron↗
Characterization of branched monounsaturated hydrocarbons of Sarcina lutea and Sarcina flava.
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Alcohols↗
[Amino acid sequence of the murein of Sarcina ventriculi and Sarcina maxima].
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Aerobiosis↗
Amino acid coding in Sarcina lutea and Saccharomyces cerevisiae.
Aminoacyl-tRNA's from Sarcina lutea were tested for incorporation into protein in a heterologous system from Escherichia coli or for biniding in a homologous system from Sarcina lutea. Aminoacyl-tRNA's from Saccharomyces cerevisiae were tested for biniding in a homologous Saccharomyces cerevisiae system. Synthetic polyribonucleotides were used as messengers. The code which exists in Sarcina lutea and Saccharomyces cerevisiae is the same as in Escherichia coli.
Amino Acids↗
Abomasal bloat associated with Sarcina-like bacteria in goat kids.
An epizootic of abdominal tympany in goat kids as a result of abomasal bloat associated with a short duration of clinical signs was fatal in over 200 kids. Histologic examination of sections of abomasum revealed high numbers of bacteria that were morphologically identical to Sarcina sp. Sarcina sp are anaerobic, gas-producing organisms that could cause abomasal bloat. Other reports have proposed that abomasal bloat is caused by abnormal abomasal flora; we propose that in the goat kids reported here, Sarcina sp may represent the abnormal flora.
Abomasum↗
Membrane lipid alkyl chain motional dynamics is conserved in Sarcina ventriculi despite pH-induced adaptative structural modifications including alkyl chain tail to tail coupling.
Sarcina ventriculi, an anaerobic Gram-positive bacterium, adapts to increasing temperature, the presence of organic solvents, or the lowering of the pH of its growth medium by joining the tails of membrane lipids from opposite sides of the bilayer, forming transmembrane, bifunctional fatty acid species. Since this is done to offset the increase in membrane mobility caused by these perturbations, it is of interest to determine whether the motional (dynamic) properties of membrane lipid alkyl chains are conserved. In this study, conservation of the motional time scales of the alkyl chains of total membrane lipids from Sarcina ventriculi cells grown at different pH values was demonstrated using proton nuclear magnetic resonance (NMR) spectroscopy. The NMR longitudinal relaxation times (T1) of the protons in the bulk methylene groups were measured for lipids from cells grown at pH 3.0 and 7.0. These measurements indicated that the temperature profile of the T1 relaxation behavior for the methylene protons from these two different preparations was the same. Analysis of the data from T1 measurements indicated that the thermal barrier for relaxation is the same in both lipid systems. This is only true if the pH of the sample on which the measurement is being made is adjusted to the same value as that at which the corresponding cells were cultured. It is clear from this latter observation that the state of protonation of the lipid head groups is a contributor to the overall motional freedom of the membrane lipid components. The correlation times (tau c) of characteristic lipid alkyl chain motion were estimated to be approximately 10(-10) s.(ABSTRACT TRUNCATED AT 250 WORDS)
Adaptation, Physiological↗
Production of coenzyme A by Sarcina lutea.
To develop an efficient method for the production of coenzyme A (CoA), optimal conditions for its formation from pantothenic acid, cysteine, and adenine were studied. A number of microorganisms were screened for production of CoA. Strains belonging to the genera Sarcina, Bacillus, Microbacterium, Micrococcus, and Serratia accumulated CoA. Among these, Sarcina lutea was selected as the best organism, and the culture conditions for the production of CoA were investigated with this organism. Under optimal conditions, 600 mug of CoA per ml was accumulated in the culture broth. CoA was readily isolated in high purity by the use of charcoal, diethylaminoethyl-cellulose, Sephadex G-25, and Dowex-50. Yields of isolated CoA were over 33% from culture broth.
Adenine↗
14-C incorporation into the fatty acids and aliphatic hydrocarbons of Sarcina lutea.
An initial investigation into the mechanism of hydrocarbon biosynthesis in Sarcina lutea was performed by measuring the amounts of (14)C incorporated into the hydrocarbons and fatty acids by use of a combination gas chromatograph and high-temperature gas-flow ionization apparatus. Uniformly labeled l-isoleucine-(14)C was predominantly incorporated into the anteiso-branched chains. Palmitate-16-(14)C gave evidence that a direct correlation may exist between the nonpolar end of the palmitate and the biosynthesis of hydrocarbons and carotenoids. The label from palmitate-1-(14)C was incorporated into the various hydrocarbon groups as a compound, derived from the polar end of the palmitate, consisting of more than two carbon atoms. Palmitate-16-(14)C and -1-(14)C gave no detectable evidence that transformed products were incorporated into other fatty acids. Sodium acetate-2-(14)C and uniformly labeled l-leucine-(14)C gave evidence of a nonspecific incorporation into both the aliphatic hydrocarbons and fatty acids of Sarcina lutea.
Acetates↗
New isoprenologs of vitamin K2 series in Sarcina lutea.
Four new menaquinone homologs have been isolated and characterized from Sarcina lutea. The use of purified silica gel G in the isolation of pure menquinones from bacterial nonpolar lipids has been developed. During this procedure, fractionation of any naturally occuring menaquinones with sidechain variation takes place. The structures of the isolated menaquinones have been elucidated by UV-, and mass spectrometry, and by their corresponding hydroquinones and layer chromatography properties. Thus, Sarcina lutea has been shown to produce four homologs of dihydromenaquinones, i.e., dihydromenaquinone-6,-7,-8, and -9.
Chromatography, Gel↗
A mathematical model explaining the molecular weights and distribution of very long chain dicarboxylic acids formed during the adaptive response of Sarcina ventriculi.
A simple mathematical model is presented to explain a recent new discovery of an unusual membrane adaptive response in Sarcina ventriculi. In this response, this organism synthesizes very long chain alpha, omega-dicarboxylic acids ranging from 28 to 36 carbon atoms in length. The distribution of chain lengths of the new fatty acid species is not consistent with de novo synthesis but suggests elaboration from the existing regular-chain fatty acids by a coupling process. Here, we demonstrate, using a mathematical model, that if the molecular weights and relative abundances of regular chain fatty acids are known, the molecular weights and relative abundances of the new, very long chain dicarboxylic fatty acid species can be predicted using a model based on the random, pairwise combination of regular chain species. This combination takes place across the bilayer leaflet to form transmembrane fatty acids. It is proposed that this coupling phenomenon is regulated by the motional dynamics of the membrane.
Cell Membrane↗
The effect of temperature on the growth and lipid composition of the extremely halophilic coccus, Sarcina marina.
Sarcina marina (NCMB 778) grew over the temperature range 20-45 degrees C but no growth was recorded at 15 degrees C or 50 degrees C. At the optimum growth temperature of 34 degrees C the doubling time was 14.5 h. The major polar lipid components, tentatively identified as the diether analogues of phosphatidyl glycerophosphate (PGP), phosphatidyl glycerol (PG), diglycosyl diglyceride (DGD) and triglycosyl diglyceride (TGD), and the major neutral lipid components, tentatively identified as squalene, dihydrosqualene, tetrahydrosqualene, vitamin MK8, geranyl geraniol and di-O-phytanyl glycerol, are identical to those found in other extremely halophilic rods and cocci. The total lipid content varied with growth conditions from 0.6-3.2% of the dry cell weight, polar lipids accounted for between 94.3 and 83.6% of the total lipid, the remainder being neutral lipid. In response to both the transition from exponential to stationary phase and a reduction of 14 degrees C in growth temperature, batch cultures showed: (i) an increase in total lipid content; (ii) a decrease in PG and (iii) an increase in PGP. Specific responses to the temperature decrease were (i) increased total lipid content; (ii) no decrease in neutral lipids in stationary phase; (iii) marked reduction in PG and (iv) raised DGD. (i) and (ii) could be mechanisms for increasing membrane fluidity. In common with all other extreme halophiles investigated the alkyl side chains of S. marina polar lipids were identified as the phytanyl (3R, 7R, 11R, 15-tetramethylhexadecyl) group. Its structure did not appear to vary with temperature so that the normal mechanisms for modifying the structure of lipid alkyl side chains to modulate membrane fluidity in response to temperature changes probably does not occur in this group of microorganisms.
Glycolipids↗
Studies on the endogenous metabolism and senescence of starved Sarcina lutea.
1. When washed suspensions of Sarcina lutea are starved aerobically in phosphate buffer at the growth temperature of 37 degrees , the rate of endogenous oxygen consumption decreases to very low values after 10hr., although many of the cells survive for 40hr. If starvation is prolonged further, the bacteria die at a rate of approximately 1.5% of the initial viable population per hour. 2. Oxidation of intracellular free amino acids accounts for most of the observed endogenous oxygen uptake but RNA is also utilized and a portion of the component bases and pentose is degraded and presumably oxidized. Ammonia appears in the supernatant and some pentose and ultraviolet-absorbing nucleotide are released from the cells. DNA, protein and polysaccharide are not measurably degraded. 3. Survival can be correlated with the ability of aerobically starved bacteria to oxidize exogenous l-glutamate and glucose. When starved under nitrogen for 40hr. cells continue to oxidize their endogenous reserves at undiminished rates when transferred to aerobic conditions; on prolonging anaerobic starvation the rate of oxidation declines during the period of most rapid loss of viability. 4. In the presence of Mg(2+), RNA degradation during aerobic starvation is almost completely suppressed without affecting the period for which the bacteria survive. 5. Cells grown in peptone supplemented with glucose accumulate reserves of polysaccharide which are metabolized in aerobic starvation, together with free amino acids. Ammonia is evolved and RNA is degraded to a greater extent than in peptone-grown suspensions. Bacteria rich in polysaccharide survive less well than those which are deficient in the polymer; the reason for this phenomenon has yet to be established. 6. In peptone medium, endogenous oxygen uptake and the concentration of intracellular free amino acids decline as growth progresses and they continue to decrease when the organism is held in stationary phase. Under the conditions used, the endogenous Q(o2) and free amino acid pool of cells grown in peptone with 2% (w/v) glucose did not decline so markedly and the bacteria contained large amounts of polysaccharide at all stages of growth.
Aging↗
Purification and characterization of pyruvate decarboxylase from Sarcina ventriculi.
Pyruvate decarboxylase from the obligate anaerobe Sarcina ventriculi was purified eightfold. The subunit Mr was 57,000 +/- 3000 as estimated from SDS-PAGE, and the native Mr estimated by gel filtration on a Superose 6 column was 240,000, indicating that the enzyme is a tetramer. The Mr values are comparable to those for pyruvate decarboxylase from Zymomonas mobilis and Saccharomyces cerevisiae, which are also tetrameric enzymes. The enzyme was oxygen stable, and had a pH optimum within the range 6.3-6.7. It displayed sigmoidal kinetics for pyruvate, with a S0.5 of 13 mM, kinetic properties also found for pyruvate decarboxylase from yeast and differing from the Michaelis-Menten kinetics of the enzyme from Z. mobilis. No activators were found. p-Chloromercuribenzoate inhibited activity and the inhibition was reversed by the addition of dithiothreitol, indicating that cysteine is important in the active site. The N-terminal amino acid sequence of pyruvate decarboxylase was more similar to the sequence of S. cerevisiae than Z. mobilis pyruvate decarboxylase.
Amino Acid Sequence↗
Production of L-serine by Sarcina albida.
Conditions for the production of microbial L-serine hydroxymethyltransferase and for the conversion of glycine to L-serine were studied. A number of microorganisms were screened for their abilities to form and accululate L-serine from glycine, and Sarcina albida was selected as the best organism. Enzyme activity in this organism as high as 0.12 U/ml could be produced in shaken cultures at 30 degrees C in a medium containing glucose, ammonium sulfate, glycine, yeast extract, and inorganic salts. L-Serine was produced most efficiently by shaking cells at 30 degrees C in a reaction mixture containing 20% glycine, 5 X 10(-3) M formaldehyde, and 3 X 10(-4) M pyridoxal phosphate in yields of 22 mg of broth in 5 days. L-Serine was easily isolated in 84% yields by ion-exchange resin.
Bacteria↗
Production of flavine-adenine dinucleotide from riboflavine by a mutant of Sarcina lutea.
A study was made to develop a new method for the production of flavine-adenine dinucleotide (FAD) from riboflavine and adenine by a mutant of Sarcina lutea deficient in the enzyme adenosine deaminase. It was found that this strain could convert exogenously supplemented riboflavine to extracellular FAD. The yields of FAD were increased by addition of D-cycloserine in the culture medium. The culture conditions for FAD production were investigated under the addition of D-cycloserine, and increased production of FAD was observed with the addition of an appropriate amount of thiamine, acetate, and sodium ion. The yield of 0.7 g/liter was obtained in the optimal culture in 5 days. Accumulated FAD was readily isolated by adsorption chromatography and ion-exchange chromatography in a 70% yield.
Acetates↗
Mechanism of excretion of a bacterial proteinase: demonstration of two proteolytic enzymes produced by a Sarcina strain (Coccus P).
A Sarcina strain (Coccus P) produces two proteolytic enzymes. One is found only extracellularly, is far more prevalent, and is actively excreted during exponential growth. It is the enzyme responsible for the known strong proteolytic activity of the cultures of this strain. A second protease is, however, produced which remains associated with the intact cells but is released by the protoplasts. The two enzymes appear unrelated in their derivation. Calcium ions play an essential role in preventing autodigestion of the excreted enzyme.
Acetates↗