[Mechanism of action of the extracellular bacteriolytic enzymes of Lysobacter sp. on gram-positive bacteria: role of the cell wall anionic polymers of the target bacteria]. / Rol' anionnykh polimerov kletochnykh stenok grampolozhitel'nykh bakterii-mishenei v mekhanizme deistviia na nikh vnekletochnykh bakterioliticheskikh fermentov Lysobacter sp.

The study of the extracellular bacteriolytic enzymes of Lysobacter sp. showed that they can efficiently hydrolyze the peptidoglycan of gram-positive bacteria provided that there is an electrostatic interaction of these enzymes with the cell wall anionic polymers, teichoic and teichuronic acids in particular. The hydrolytic action of bacteriolytic enzymes on the cell wall largely depends on the negative charge of teichoic and teichuronic acids, rather than on their chemical composition.

Cell wall teichoic acids of two Brevibacterium strains.

Structurally identical teichoic acids were detected in cell walls of two soil isolates assigned to Brevibacterium linens based on phylogenetic data. Both cell walls contain unsubstituted 1,3-poly(glycerol phosphate) and poly(glycosylglycerol phosphate). Repeating units of the latter--alpha-D-GlcpNAc-(1-->4)-beta-D-Galp-(1-->1)-Gro--are bound by phosphodiester bonds including OH-3 of galactose and OH-3 of glycerol. Some of the N-acetylglucosamine residues have 4,6-pyruvic acid acetal, amounts of the latter in the two strains being unequal. Species-specificity of the structures of teichoic acids in the genus Brevibacterium is discussed.

Structures of cell wall teichoic acids of Brevibacterium iodinum VKM Ac-2106.

Structures of two cell wall teichoic acids of Brevibacterium iodinum VKM Ac-2106 were studied. The structure of mannitol teichoic acid described earlier was mainly confirmed. This polymer is 1,6-poly(mannitol phosphate) bearing beta-D-glucopyranosyl residues at the C-2 of mannitol and pyruvic acid residues at the C-4 and C-5. The absolute configurations of D-mannitol and S-pyruvic acid were found. The following distinctions from the earlier described structure were found: unsubstituted 1,6-poly(mannitol phosphate) residues and residues substituted only by beta-D-glucopyranosyl at the C-2 of mannitol but unsubstituted by pyruvic acid are present in the chain. The structure of glycerol teichoic acid present in the cell wall as a minor component (approximately 7%) is also described. This acid is identified as 1,3-poly(glycerol phosphate) substituted at the C-2 of glycerol by 2-acetamido-2-deoxy-alpha-D-galactopyranosyl residues bearing R-pyruvic acid residues at the C-4 and C-6 of galactose. This polymer is for the first time described in the cell wall of Gram-positive bacteria.

[The cell wall teichoic acids of streptomycetes from the "Streptomyces cyaneus" cluster]. / Teikhoevye kisloty kletochnykh stenok streptomitsetov klastera "Streptomyces cyaneus".

The cell wall anionic polymers of the 13 species of the "Streptomyces cyaneus" cluster have a similar structure and contain beta-glucosylated 1,5-poly(ribitol phosphate) and 1,3-poly(glycerol phosphate). In the degree of glucosylation of the ribitol phosphate units of their teichoic acids, the cluster members can be divided into two groups. The streptomycetes of the first group (S. afghaniensis, S. janthinus, S. purpurascens, S. roseoviolaceus, and S. violatus) are characterized by a very similar structure of their cell walls, completely glucosylated 1,5-poly(ribitol phosphate) chains, and a high degree of DNA homology (67-88%). The cell wall teichoic acids of the second group (S. azureus, S. bellus, S. caelestis, S. coeruleorubidus, S. curacoi, and S. violarus) differ in the degree of beta-glucosylation of their 1,5-poly(ribitol phosphate) chains and have a lower level of DNA homology (54-76%). Two streptomycetes of the cluster (S. cyaneus and S. hawaiiensis) are genetically distant from the other cluster members but have the same composition and structure of the cell wall teichoic acids as the second-group streptomycetes. The data obtained confirm the genetic relatedness of the "S. cyaneus" cluster members and suggest that the structure of the cell wall teichoic acids may serve as one of the taxonomic criteria of the species-level status of streptomycetes.

[Teichoic acids in the cell walls of Microbispora mesophila Ac-1953t and Thermobifida fusca Ac-1952t]. / Teikhoevye kisloty kletochnykh stenok Microbispora mesophila Ac-1953t i Thermobifida fusca Ac-1952t.

The cell walls of Microbispora mesophila strain Ac-1953T (the family Streptosporangiaceae) and Thermobifida fusca Ac-1952T (the family Nocardiopsiceae) were found to contain teichoic acids of a poly(glycerol phosphate) nature. The teichoic acid of M. mesophila (formerly Thermomonospora mesophila) represents a poly(glycerol phosphate) containing 5% of substituent 2-acetamido-2-deoxy-alpha-galactosaminyl residues. The teichoic acid of such kind was found in actinomycetes for the first time. The cell wall of T. fusca (formerly Thermonospora fusca) contains two teichoic acids, namely, unsubstituted 1,3-poly(glycerol phosphate) and beta-glucosylated 1,3-poly(glycerol phosphate).

Cell wall teichoic acids of actinomycetes of three genera of the order actinomycetales.

The structures of cell wall teichoic acids of the members of newly recognized genera of the order Actinomycetales were studied. Planotetraspora mira VKM Ac-2000T contains two types of teichoic acids: 2,3-poly(glycerol phosphate) substituted with alpha-D-Galp at C-1 of glycerol and 1,3-poly(glycerol phosphate) substituted with alpha-L-Rhap at OH-2 of glycerol (60%). Herbidospora cretacea VKM Ac-1997T contains the chains of 1,3-poly(glycerol phosphate) partially substituted with alpha-D-Galp and alpha-D-GalpNAc at C-2 of glycerol. The majority of alpha-D-galactopyranosyl residues are substituted at OH-3 with a sulfate. The aforementioned teichoic acids have not been found in bacteria thus far. Actinocorallia herbida VKM Ac-1994T contains poly(galactosylglycerol phosphate), with the beta-Galp-(1-->2)-Gro-P repeating units being linked via the phosphodiester bonds between the OH-3 of glycerol and OH-6 of galactose. Earlier, this structure was found in the cell wall of Actinomadura madura. The polymer structures were determined by chemical analysis and using 13C-NMR spectroscopy. The results show that teichoic acids are widespread in the order Actinomycetales.

NMR-based identification of cell wall anionic polymers of Spirilliplanes yamanashiensis VKM Ac-1993(T).

The cell wall of Spirilliplanes yamanashiensis VKM Ac-1993(T) contains four anionic polymers, viz., three teichoic acids and a sugar-1-phosphate polymer. The following are the structures of the teichoic acids: poly[-6-beta-D-glucopyranosyl-(1-->2)-glycerol phosphate] (PI), 1,3-poly(glycerol phosphate) bearing N-acetyl-alpha-D-glucosamine residues at O-2 (70%) (PII), and poly[-6-N-acetyl-alpha-D-glucosaminyl-(1-->2)-glycerol phosphate] (PIII). The repeating unit of the fourth polymer (PIV) has the structure of -6-alpha-D-GlcpNAc-(1-->6)-alpha-D-GlcpNAc-1-P- with a 3-O-methyl-alpha-D-mannopyranosyl residues at position 3 of some 6-phosphorylated N-acetylglucosamine residues (50%). Polymers PI, PIII and PIV have not hitherto been found in prokaryotic cell walls.

[The carbohydrate-containing cell wall polymers of some species from the cluster "Streptomyces lavendulae"]. / Uglevodsoderzhashchie polimery kletochnykh stenok nekotorykh streptomitsetov, vkhodiashchikh v klaster "Streptomyces lavendulae".

The type strains of the cluster "Streptomyces lavendulae" species with a low level of DNA-DNA relatedness were found to contain different cell-wall carbohydrate polymers, whereas the species of this cluster with a level of DNA-DNA relatedness of about 60% contain similar or identical carbohydrate polymers. The type strains Streptomyces katrae VKM Ac-1220T and S. polychromogenes VKM Ac-1207T synthesize mannan with different amounts of alpha-1,2- and alpha-1,3-substituted mannopyranose units and a small number of 1,3-poly(glycerophosphate) chains. The cell walls of S. lavendulocolor VKM Ac-215T and Streptomyces sp. VKM Ac-2117 were found to contain a hitherto unknown teichuronic acid, whose repeating unit is a disaccharide consisting of diaminomannuronic acid and N-acetylgalactosamine:-->4)-beta-D-ManpNAc3NAcA-(1-->3)-alpha-D- GalpNAc-(1-->. In addition, the cell walls of these two streptomycetes contain beta-glucosylated 1,5-poly(ribitol phosphate). The cell walls of S. virginiae VKM Ac-1218T and S. flavotricini VKM Ac-1277T contain the same poly(glucosyl-glycerophosphate). The results presented in this paper are in accordance with the DNA-DNA relatedness data published earlier and indicate a taxonomic significance of the structure of the cell-wall polysaccharides for the delineation of pheneticall/related Streptomyces species.

Cell wall teichoic acids: structural diversity, species specificity in the genus Nocardiopsis, and chemotaxonomic perspective.

Data on the structures of cell wall teichoic acids, the anionic carbohydrate-containing polymers, found in many Gram-positive bacteria have been summarized and the polymers of the actinomycete genus Nocardiopsis have been considered from the taxonomic standpoint. The structures of these polymers or their combinations have been demonstrated to be indicative of each of seven Nocardiopsis species and two subspecies, verified by the DNA-DNA relatedness data, and to correlate well with the grouping of the organisms based on 16S rDNA sequences. As each of the intrageneric taxa discussed is definable by the composition of teichoic acids, the polymers are considered to be valuable taxonomic markers for the Nocardiopsis species and subspecies. The (13)C NMR spectra of the polymers, data on the products of their chemical degradation, and distinguishing constituents of whole cell walls derived from teichoic acids are discussed, which are useful for identification of certain polymers and members of the genus Nocardiopsis at the species and subspecies level in microbiological practice.

Structure of anionic carbohydrate-containing cell wall polymers in several representatives of the order actinomycetales.

A new teichoic acid was identified in the cell walls of Streptomyces griseoviridis VKM Ac-622T, Streptomyces sp. VKM Ac-2091, and Actinoplanes campanulata VKM Ac-1319T. The polymer is poly(glycosylglycerol phosphate). The repeating units of the polymer, alpha-galactopyranosyl-(1-->3)-2-acetamido-2-deoxy-beta-galactopyran+ ++ osyl-(1-->1)-glycerols, are in phosphodiester linkage at C-3 of glycerol and C-6 of galactose. The structures of cell wall teichoic acids in the strains Streptomyces chryseus VKM Ac-200T and "Streptomyces subflavus" VKM Ac-484 similar in morphology and growth characteristics are also identical: 1,5-poly(ribitol phosphate) substituted at C-4(2) by 2-acetamido-2-deoxy-beta-glucopyranosyl residues and 1,3-poly(glycerol phosphate). The taxonomic aspects of these results are discussed.

[Teichoic acids from the cell wall of Nocardiopsis prasina BKM Ac-1880T]. / Teikhoevye kisloty kletochnoi stenki Nocardiopsis prasina BKM Ac-1880T.

The cell wall of Nocardiopsis prasina VKM Ac-1880T was found to contain two structurally different teichoic acids: unsubstituted 3,5-poly(ribitol phosphate) and 1,3-poly(glycerol phosphate), substituted at position 2 by 10% with alpha-N-acetylglucosamine and by 5% with O-acetyl groups. The structure of the polymers was studied by chemical analysis and NMR spectroscopy. The results obtained correlate well with 16S rRNA sequence data and confirm the species-specificity of teichoic acids in the genus Nocardiopsis.

Structure of a teichoic acid from Nocardioides luteus VKM Ac-1246T cell wall.

A teichoic acid from the cell walls of Nocardioides luteus VKM Ac-1246T, a validly described species of the Nocardioides genus, is a 1,5-poly(ribitol phosphate) completely substituted at C-4 by alpha-D-galactopyranosyl residues carrying a 4,6-pyruvate ketal group in R-configuration. The structure of the repeating unit of the polymer is as follows: [figure]. The chain consists of approximately 18 repeating units and six beta-D-galactofuranosyl residues linked in the oligomer by 1,6-glycosidic bonds. The oligomer probably terminates the growing end of the teichoic acid. The structure of the polymer was determined by chemical methods and NMR spectroscopy. This teichoic acid has not been described so far.

A teichoic acid of Nocardioides albus VKM Ac-805T cell walls.

A teichoic acid of Nocardioides albus VKM Ac-805T cell walls, a typical species of the genus Nocardioides, contains a poly(glycosylglycerol phosphate). The repeating unit of the polymer has the structure: [figure]. These units are in phosphodiester linkage at C-3 of glycerol and C-3 of beta-D-galactopyranose. beta-D-Galactopyranosyl residues are substituted at C-4 by beta-D-glucopyranose carrying a 4,6-pyruvate ketal group in S-configuration. The presence of pyruvic acid in the majority of repeating units increases the anionic properties of the polymer in comparison with most other common teichoic acids. This is the first report of the occurrence of a beta-D-galactofuranosyl residue in teichoic acids; it probably acts as a terminator of an extending chain of the polymer. The ratio of beta-D-galactopyranosyl to beta-D-galactofuranosyl units is 7:1. The polymer structure was determined by NMR spectroscopy. This type of teichoic acid structure has not been reported previously.

Anionic carbohydrate-containing polymers of cell walls in two streptoverticille genospecies.

The cell walls of two streptoverticille genospecies which belong to a historically isolated group of the genus Streptomyces contain anionic polymers of different structure. Streptomyces hachijoensis VKM Ac-191T and Streptomyces cinnamoneus subsp. azacoluta VKM Ac-606T assigned to one genospecies on the basis of DNA--DNA hybridization [5] contain 37% of an identical sugar-1-phosphate polymer. The repeating disaccharide units of the polymer, 2-amino-2-deoxy-alpha-D-glucopyranosyl-(1-->6)-2-acetamido-2-deoxy-al pha-D-glucopyranose, are linked at C-1 and C-6' by phosphodiester bonds. The cell walls of Streptomyces biverticillatus VKM Ac-891T and Streptomyces baldaccii VKM Ac-821T, members of another genospecies, contain about 30% 1,3-poly(glycerol phosphate) completely substituted by 2-amino-2-deoxy-alpha-D-glucopyranosyl residues at C-2. Due to the presence of an amino sugar with a free amino group in the repeating unit, the polymers exhibit neutral properties. Polymer structures were determined by chemical methods and NMR spectroscopy. The data indicate taxonomic specificity of anionic polymers in streptoverticille cell walls.

Cell wall teichoic acids of Actinomadura viridis VKM Ac-1315T.

The cell walls of Actinomadura viridis contain poly(glycosylglycerol phosphate) chains of complex structure. On the basis of NMR spectroscopy of the polymer and glycosides thereof the following structural units were found: beta-D-Galp3Me-(1-->4)[beta-D-Glcp-(1-->6)]-beta-D-Galp-(1-->1)-++ +snGro (G1); beta-D-Galp-(1-->4)-beta-D-Galp-(1-->1)-snGro (G2); beta-D-Galp3Me-(1-->4)-beta-D-Galp-(1-->1)-snGro (G2a); beta-D-Galp-(1-->1)-snGro (G3); beta-D-Galp-(1-->1)[beta-D-Galp-(1-->2)]-snGro (G4); beta-D-Glcp-(1-->2)-snGro (G5). Glycosides G1, G2 and G3 were the predominant components of the teichoic acid: they formed the polymer chain via phosphodiester bonds involving C-3 of the glycerol residue and C-3 of the galactosyl residue which in turn glycosylates C-1 of the glycerol residue. Whether the different glycosides make up the one chain or whether there are several poly(glycosylglycerol phosphate) chains in the cell wall remains to be determined. It was suggested that the minor component G5 is located at the nonterminal end of the chains. Compound G4 which contains disubstituted glycerol residues (unusual for the teichoic acid) was also found as a minor component; this may be the glycoside of a new type of teichoic acid, or a glycoside on the terminal end of the above mentioned chains. In addition, small amounts of 1,3-poly(glycerol phosphate) chains were found in the cell wall.

Structure of teichoic acid of the cell wall of Streptomyces sparsogenes VKM Ac-1744T.

The cell wall of Streptomyces sparsogenes VKM Ac-1744T contains a teichoic acid of 1,3-poly(glycerol phosphate) type. The polymer is composed of about 11 repeating units of the following structure: The structure was not identified in gram-positive bacteria previously. The data are in agreement with DNA-relatedness between Streptomyces sparsogenes and Streptomyces hygroscopicus species and suggest the species-specificity of teichoic acid structure in the studied streptomycetes group.

Teichoic acids of the cell wall of Nocardiopsis listeri, Nocardiopsis lucentensis, and Nocardiopsis tregalosei.

The structures of teichoic acids of three Nocardiopsis species were established. The cell wall of Nocardiopsis listeri VKM Ac-1881T contains two teichoic acids (TA). TA1 is 1,3-poly(glycerol phosphate) with 50% glycerol phosphate residues substituted by alpha-N-acetylglucosamine in the C-2 position. TA2 is 1, 5-poly(ribitol phosphate) with each ribitol phosphate unit carrying pyruvate acetal groups in the 2 and 4 positions. The cell wall of Nocardiopsis lucentensis VKM Ac-1963T contains only one teichoic acid of the same structure as TA1 of N. listeri. The teichoic acid of the Nocardiopsis tregalosei VKM Ac-942 is a 1,3-poly(glycerol phosphate) substituted (60%) by beta-glucopyranosyl residues. Structures of polymers were studied by chemical and NMR spectroscopy methods. The presented results confirm the species-specificity of teichoic acids from Nocardiopsis genus.

Anionic polymers in cell walls of gram-positive bacteria.

Information on the prevalence, compositions, and structures of anionic carbohydrate-containing polymers of cell walls of Gram-positive bacteria is summarized. The data suggest that these polymers are important for normal functioning of bacterial cells and require further studies. Structural data on teichoic acids found in the literature published over the last few years are discussed. This is a very diverse class of polymers whose structure-specific pathways of degradation were studied and NMR spectra were examined. Unique comprehensive tables of 13C-NMR spectroscopic data (mainly obtained by the authors) on these polymers are given in the Appendix. Other tables summarize data on teichuronic acids, sugar-phosphate polymers, acid polysaccharides, and structural variants of bonds between acid polysaccharides and peptidoglycans known from the literature. Functions of anionic polymers and their possible chemotaxonomic applications are discussed.

[Lytic action of lysoamidase from Xanthomonas sp. correlates with the presence of the target ribitol teichoic acids in the cell wall of gram-positive bacteria]. / Liticheskoe deistvie lizoamidazy iz Xanthomonas sp. korreliruet s prisutstviem v kletochnoi stenke grampolozhitel'nykh bakterii--mishenei ribitteikhoevykh kislot.

Lysoamidase, a bacteriolytic complex from the culture liquid of Xanthomonas sp., hydrolyzed the cells walls of Staphylococcus aureus, Streptomyces chrysomallus, and Streptomyces azureus, which contain ribitol teichoic acids in addition to peptidoglycan. The cell walls of Streptomyces roseoflavus, Glycomyces harhinensis, and Nocardiopsis dassonvillei, containing glycerol teichoic acids, were not hydrolyzed by lysoamidase. The extent of the hydrolysis of 20-h Str. chrysomallus cells and cell walls, containing 40% ribitol teichoic acids, was considerably higher than that of 40-h cells and cell walls, containing 15% teichoic acids. Homogeneous bacteriolytic enzymes of the lysoamidase complex (muramidase and two bacteriolytic peptidases) most efficiently hydrolyzed S. aureus and Str. chrysomallus cell walls, characterized by the highest content of ribitol teichoic acids, and did not hydrolyze purified peptidoglycan.

Poly(arabitol phosphate) teichoic acid in the cell wall of Agromyces cerinus subsp. cerinus VKM Ac-1340T.

On the basis of NMR studies and analysis of the products of acid and alkaline hydrolyses the following structures were established for the repeating units of poly(arabitol phosphate) teichoic acid: alpha-6-deoxy-L-Talp-(1-->3)-beta-D-GLcpNAc-(1-->2)-alpha-L-Rha p-(1-->4(2)-D- Arabitol-PO4 and beta-D-GlcpNAc-(1-->2)-alpha-L-Rhap-(1-->4(2)-D-arabitol-PO4. The molar ratio of these units is about 1.2:1.0, respectively. Poly(arabitol phosphate) teichoic acid is here reported in bacterial cells walls for the first time.