[Ecology of fermentation sarcinas Sarcina ventriculi and Sarcina maxima].
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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.
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.
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A study of abomasal disease in lambs aged 2-5 weeks, made during the period 1993-1998, included 67 cases and 45 non-affected controls. Gross pathological findings included various combinations of bloat, haemorrhage and ulcers in the abomasum. Sarcina -like bacteria were found in sections and smears from the abomasum of 79% (53/67) of the cases. From one case, a lamb with abomasal bloat, the anaerobic "packet"-forming Sarcina ventriculi was cultivated from the abomasal contents and identified by biochemical reactions and sequencing of the 16S rRNA gene. Sarcina -like bacteria were observed microscopically in specimens from 94% (44/47) of the lambs with abomasal gas and in 45% (9/20) of those with ulcers or haemorrhage or both but little gas. On culture, abomasal contents from 41 cases yielded Clostridium fallax from 16 (39%) and Clostridium sordellii from eight (20%); abomasal cultures from 30 control lambs were negative for the three bacterial species. Quantitative cultivation, carried out on abomasal contents from live lambs and lambs dead </=3 h, showed that Clostridium perfringens, Escherichia coli and Lactobacillus spp. were present in the majority of both cases and controls, with no significant differences in the mean numbers.
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.
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.
Mathews, Micheline M. (Department of Bacteriology, University of California, Berkeley). Comparative study of lethal photosensitization of Sarcina lutea by 8-methoxypsoralen and by toluidine blue. J. Bacteriol. 85:322-328. 1963.-A comparative study has been made of the photo-killing of Sarcina lutea by 8-methoxypsoralen (8-MOP) and by toluidine blue. It has been found that photosensitization by 8-MOP differs from photosensitization by toluidine blue, in that it has a temperature coefficient of less than one, and that the presence of oxygen is not necessary for, and even is deleterious to, the photosensitization, the psoralen being destroyed by its presence. It has previously been shown that the presence of carotenoid pigments protects the cells of S. lutea from lethal photosensitization by toluidine blue; it was found that the presence of these pigments has no protective effect in photosensitization with 8-MOP. Studies on the lethal photosensitization of S. lutea with toluidine blue suggested that the primary sensitive site of the photokilling was the protein of the cell membrane, as manifested by the destruction of membrane enzyme activity and the regulation of permeability. It has been found that photokilling by 8-MOP has no effect on these functions. A study was made on the effect of photokilling by 8-MOP on the production of penicillin-resistant mutants as an indication of an alteration in the cellular deoxyribonucleic acid (DNA) by the psoralen. Psoralen photosensitization resulted in the development of many penicillin-resistant mutants. On the basis of the findings reported in this paper, it is suggested that photosensitization of S. lutea by 8-MOP does not reflect damage to cellular protein, as does toluidine blue, but rather damage to cellular DNA.
Huston, Charles K. (U.S. Army Biological Laboratories, Fort Detrick, Frederick, Md.), and Phillip W. Albro. Lipids of Sarcina lutea. I. Fatty acid composition of the extractable lipids. J. Bacteriol. 88:425-432. 1964.-The extractable lipids of Sarcina lutea were separated into several fractions by a combination of column and thin-layer chromatography. Qualitative and quantitative characterization of the fatty acid content of these lipid fractions was accomplished by means of gas-liquid chromatography and infrared analyses. Of the total extract, the lipids consisted of 2.1% free fatty acids, 51.0% glycerides, and 22.7% complex lipids; they had a fatty acid content with a complete spectrum of carbon numbers from C(8) to C(22). The fatty acids included a large component of branched-acids in addition to the normal straight-chain acids. The branched-acids, comprising 40% of the fatty acids analyzed, constituted a homologous series of iso-acids from C(12) to C(19). Two 18-carbon unsaturates were found cis-9-octadecenoate and cis-11-octadecenoate. A relatively high percentage (20.5%) of the extractable material from S. lutea was found to be hydrocarbon. This material was not further characterized.
Albro, Phillip W. (Ft. Detrick, Frederick, Md.), and Charles K. Huston. Lipids of Sarcina lutea. II. Hydrocarbon content of the lipid extracts. J. Bacteriol. 88:981-986. 1964.-The hydrocarbon fraction from Sarcina lutea lipid extracts was characterized by a combination of thin-layer and gas-liquid chromatography and infrared spectroscopy. A total of 37 components were observed by gas-liquid chromatography of this material. A breakdown of the components into classes indicated a composition consisting of 88.9% n-saturates, 1.2% monoenes, 2.1% dienes, 5.0% trienes, and 0.6% branched-saturates. Less than 0.1% of the hydrocarbon material was aromatic. No attempt was made in this study to relate the composition to either origin or function in the cell.
Huston, Charles K. (Fort Detrick, Frederick, Md.), Phillip W. Albro, and Gerald B. Grindey. Lipids of Sarcina lutea. III. Composition of the complex lipids. J. Bacteriol. 89:768-775. 1965.-The complex lipids from a strain of Sarcina lutea were isolated and separated into fractions on diethylaminoethyl cellulose acetate and silicic acid columns. These fractions were monitored in several thin-layer chromatography systems. The various lipid types were characterized by their behavior in thin-layer systems and by an analysis of their hydrolysis products. The fatty acid composition of the column fractions was determined by gas-liquid chromatography. A number of components (13) were separated by thin-layer chromatography and characterized. The major components were polyglycerol phosphatide (17.0%), lipoamino acids (15.1%), phosphatidyl glycerol (13.8%), and an incompletely characterized substance (15.0%). Minor constituents included phosphatidyl inositol (5.5%), phosphatidic acid (4.2%), phosphatidyl serine (2.0%), and phosphatidyl choline (1.0%). No phosphatidyl ethanolamine was observed.
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)
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.
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.
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.
When Sarcina lutea is used as test organism in a cup-plate assay, the dose-response curves for the assay of penicillin in sera of humans and rabbits are linear with the square of the zone diameter and log concentration of penicillin and parallel to the dose-response curve for penicillin in phosphate buffer. Suitable conditions for such assays are described, using either large square assay-plates and fish-spine beads or conventional Petri dishes and cylinders. The assay using large plates and fish-spine beads is much more accurate and economical of effort and serum. An assay method suitable for inclusion in the blood-level duration test for oily injection of procaine penicillin (International Pharmacopoeia) is described. The method makes it possible for fiducial limits to be readily computed for the potencies obtained.
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.