Separation of the poly(glycerophosphate) lipoteichoic acids of Enterococcus faecalis Kiel 27738, Enterococcus hirae ATCC 9790 and Leuconostoc mesenteroides DSM 20343 into molecular species by affinity chromatography on concanavalin A.

This study shows for the first time microheterogeneity of 1,3-linked poly(glycerophosphate) lipoteichoic acids. The lipoteichoic acids investigated were those of Enterococcus faecalis Kiel 27738 (I), Enterococcus hirae (Streptococcus faecium) ATCC 9790 (II), and Leuconostoc mesenteroides DMS 20343 (III). Lipoteichoic acids II and III are partially substituted by mono-, di-, tri-, and tetra-alpha-D-glucopyranosyl residues with (1----2) interglycosidic linkages. Lipoteichoic acid I is substituted with alpha-kojibiosyl residues only. Lipoteichoic acids I and III additionally carry D-alanine ester. Lipoteichoic acids were separated on columns of concanavalin-A-Sepharose according to their increasing number of glycosyl substituents per chain. It was evident that all molecular species are usually glycosylated and that alanine ester and glycosyl residues occur on the same chains. The chain lengths of lipoteichoic acid I and II vary between 9-40 glycerophosphate residues, whereas those of lipoteichoic acid III appear to be uniform (33 +/- 2 residues). Molecular species differ in the extent of glycosylation but their content of alanyl residues is fairly constant. All lipoteichoic acids contain a small fraction (5-15%) different in composition from the bulk and most likely reflecting an early stage of biosynthesis. Two procedures for chain length determination of poly(glycerophosphate) lipoteichoic acids are described.

Immunochemical studies on levans from several strains of Actinomyces viscosus.

High molecular-weight levans elaborated by 8 separate strains of Actinomyces viscosus were purified: the inteactions of these levans with concanavalin A and anti-fructan myeloma immunoglobulins UPC-10 and J606 were examined by the quantitative precipitin method. Oligosaccharides released from the levans by partial acid hydrolysis were separated by partition chromatography on paper and characterized in situ by selective spray reagents. The liberated oligomers were compared with oligomers of known structure released from levans of Aerobacter levanicum and Leuconostoc mesenteriodes B512 as well as with plant inulin. The fragmentation analysis indicated a structure for Actinomyces levans comprising chains of beta (2----6)-linked fructofuranosyl units joined through multiple (1,2,6)-linked fructosyl branch units.

Use of 13C-n.m.r. spectroscopy for the quantitative estimation of 3-O- and 3,6-di-O-substituted D-glucopyranosyl residues in alpha-D-glucans formed by the D-glucosyltransferases of Streptococcus sobrinus.

The 13C-n.m.r. spectra of the three alpha-D-glucans from Streptococcus sobrinus and the dextran from Leuconostoc mesenteroides, which differ widely in the ratios of omega (terminal, nonreducing) D-glucopyranosyl groups: 3-:6-:3,6-linked D-glucopyranosyl (Glc) residues, were measured in 0.5M NaOH at 22 degrees. The C-1 signals of 3-O-substituted Glc in a linear sequence, 6-O-substituted Glc in a linear sequence, 3,6-di-O-substituted Glc in a (1----6)-linked sequence, and Glc attached to O-3 of 3,6-di-O-substituted Glc were distinguished from each other. The C-3 signal of 3,6-linked Glc appeared downfield by 0.6 to 1.0 p.p.m. compared to the C-3 signal of 3-linked Glc in a linear sequence. The C-6 signals of omega-terminal, 3-linked, 6-linked, and 3,6-linked Glc were also assigned. The C-2 signal of 3-linked Glc in a linear sequence appeared separately, at 73.76 p.p.m. Based on these assignments, the various D-glucopyranosyl residues of the S. sobrinus alpha-D-glucans were quantitatively estimated from the signal areas of the C-2 atom of 3-linked Glc, the C-3 atom of 3-linked and 3,6-linked Glc, the C-6 atom of 6-linked and 3,6-linked Glc, and the C-6 atom of the omega-Glc groups and 3-linked Glc residues. The figures thus derived for the linkage ratios were close to those obtained by methylation analysis.

Levels of water-soluble vitamins in methanogenic and non-methanogenic bacteria.

The levels of seven water-soluble vitamins in Methanobacterium thermoautotrophicum, Methanococcus voltae, Escherichia coli, Bacillus subtilis, Pseudomonas fluorescens, and Bacteroides thetaiotaomicron were compared by using a vitamin-requiring Leuconostoc strain. Both methanogens contained levels of folic acid and pantothenic acid which were approximately two orders of magnitude lower than levels in the nonmethanogens. Methanobacterium thermoautotrophicum contained levels of thiamine, biotin, nicotinic acid, and pyridoxine which were approximately one order of magnitude lower than levels in the nonmethanogens. The thiamine level in Methanococcus voltae was approximately one order of magnitude lower than levels in the nonmethanogens. Only the levels of riboflavin (and nicotinic acid and pyridoxine in Methanococcus voltae) were approximately equal in the methanogens and nonmethanogens. Folic acid may have been present in extracts of methanogens merely as a precursor, by-product, or hydrolysis product of methanopterin.

[Macrophage activation by polysaccharides from yeast-like fungi]. / Aktivatsiia makrofagov polisakharidami iz drozhzhepodobnykh gribov.

The effect of polysaccharides isolated from yeast-like fungi on diverse activities of murine peritoneal macrophages was studied in experiments in vitro. Yeast polysaccharides had a nonspecific activating effect on macrophages in tissue culture. They enhance the adhesion of macrophages to glass, stimulate phagocytosis and intracellular digestion, and change the chemotactic activity. The activity of polysaccharides directly correlates with their molecular weight. There is a relationship between the biological activity and chemical structure of polysaccharides. The beta-structural polysaccharides are capable of activating the functions of macrophages, while beta-structural dextrans do not possess such properties.

Lipoquinones of some bacteria and mycoplasmas, with considerations on their functional significance.

In a comparative study the lipoquinones of some chemoorganotrophic, facultatively aerobic bacteria, and representative Acholeplasma, Mycoplasma, Spiroplasma, and Thermoplasma strains were investigated. The quinones were partly purified by preparative thin layer chromatography of lipid extracts, and characterized by their difference spectra (reduced minus oxidized) and Rf values. Respiring bacteria expectedly contained benzoquinones and/or naphthoquinones in micromolar concentrations whereas some aerotolerant, cytochrome-less, gram-positive bacteria were found to contain menaquinones in nanomolar concentrations, or even no quinones; only Streptococcus faecalis, an organism supposed to use a rudimentary, flavin-terminated respiratory chain system produced desmethyl menaquinone in amounts ranging between "high" and "low" quinone contents. Among the mycoplasmas investigated, only Thermoplasma acidophilum was found to be capable of synthesizing quinones (MK-7) in the micromolar order of magnitude indicating a respiratory electron transport system. The presence of energetically useful respiratory chain systems in Acholeplasma, Mycoplasma, and Spiroplasma is questioned since these organisms contain quinones (MK-4) in nanomolar concentrations, or no quinones, depending on the presence of exogeneous MK-6 in the growth medium. The possible metabolite role of menaquinones present in "low" amounts, as well as the role of NADH oxidase systems more or less tightly bound to the cytoplasmic membrane with the mycoplasmas deserves further investigation.

[Use of a freeze-drief culture of Leuconostoc mesenteroides for the synthesis of dextran]. / Ispol'zovanie dlia sinteza dekstrana liofil'no vysushennoi kul'tury Leuconostos mesenteroides

The influence of a prolonged storage of the freeze-dried culture Leuconostoc mesenteroides strain SF-4 on its capacity for the dextrane synthesis has been studied. The culture has maintained its normal viability for 12 years (the observation time) without changes in the cell morphology and capacity for the dextrane synthesis. The structure of dextrane synthesized by the revived culture L. mesenteroides has been similar to that of dextrane synthesized by the culture L. mesenteroides preserved via passages.

Cell wall constituents of Leuconostoc citrovorum and Leuconostoc mesenteroides.

The cell wall constituents of Leuconostoc citrovorum 8082, L. mesenteroides 10830a, and L. mesenteroides 11449 have been ascertained. All three strains contained glycerol. Glucose and rhamnose were the major reducing sugar constituents. Alanine, glutamic acid, lysine, glucosamine, and muramic acid were the principal amino acids and amino sugars in all three strains. In addition, strain 10830a contained l-serine as a major cell wall component. Quantitative amino acid analyses indicate that glutamic acid, lysine, glucosamine, muramic acid, and serine may be present in the cell walls in equimolar amounts and that alanine is present in three to four times these quantities. The similarities and differences between the cell wall constituents of the leuconostocs and those of the lactobacilli and streptococci are discussed.