Anionic carbohydrate-containing cell wall polymers of <Emphasis Type="Italic">Streptomyces melanosporofaciens</Emphasis> and related species

The structures of cell wall anionic carbohydrate-containing polymers in Streptomyces melanosporofaciens VKM Ac-1864^T and phylogenetically close organisms—S. hygroscopicus subsp. hygroscopicus VKM Ac-831^T, S. violaceusniger VKM Ac-583^T, S. endus VKM Ac-1331^T, S. endus VKM Ac-129, and S. rutgersensis subsp. castelarensis VKM Ac-832^T—have been comparatively studied by chemical and NMR spectroscopic methods. The natural polymer of a new, previously unknown structure, Kdn (3-deoxy-D-glycero-D-galacto-non-2-ulopyranosonic acid) with β-galactose residues at C-9, has been found in the cell walls of all the strains under study. The cell walls of all the studied organisms contain three teichoic acids (TA): a predominant TA (1,3-poly(glycerol phosphate) with N-acetylated α-glucosaminyl substitutes by C-2 of glycerol, and minor TAs, 1,3-and 2,3-poly(glycerol phosphate) polymers without substitution. Their chains have O-acetyl and O-lysyl groups. Microorganisms of the above-mentioned species differ in the number of α-glucosaminyl substitutes and in the degree of their acetylation in the predominant teichoic acid.     

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.     

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.     

Carbohydrate-containing polymers of the cell wall of the thermophilic streptomycete Streptomyces thermoviolaceus subsp. thermoviolaceus VKM Ac-1857T

Anionic polymers of the cell surface of a thermophilic streptomycete were investigated. The cell wall of Streptomyces thermoviolaceus subsp. thermoviolaceus VKM Ac-1857(T) was found to contain polymers with different structure: teichoic acid--1,3-poly(glycerol phosphate), disaccharide-1-phosphate polymer with repeating unit -6)-alpha-Galp-(1-->6)-alpha-GlcpNAc-P-, and polysaccharide without phosphate with repeating unit -->6)-alpha-GalpNAc-(1-->3)-beta-GalpNAc-(1-->. Disaccharide-1-phosphate and polysaccharide without phosphate have not been described earlier in prokaryotic cell walls.     

New structures and composition of cell wall teichoic acids from Nocardiopsis synnemataformans,Nocardiopsis halotolerans,Nocardiopsis composta and Nocardiopsis metallicus: a chemotaxonomic value

The structures of the cell wall teichoic acids (TA) from some species of the genus Nocardiopsis were established by chemical and NMR spectroscopic methods. The cell walls of Nocardiopsis synnemataformans VKM Ac-2518^T and Nocardiopsis halotolerans VKM Ac-2519^T both contain two TA with unique structures—poly(polyol phosphate-glycosylpolyol phosphate)—belonging to the type IV TA. In both organisms, the minor TA have identical structures: poly(glycerol phosphate-N-acetyl-β-galactosaminylglycerol phosphate) with the phosphodiester bond between C-3 of glycerol and C-4 of the amino sugar. This structure is found for the first time. The major TA of N. halotolerans has a hitherto unknown structure: poly(glycerol phosphate-N-acetyl-β-galactosaminylglycerol phosphate), the N-acetyl-β-galactosamine being acetalated with pyruvic acid at positions 4 and 6. The major TA of N. synnemataformans is a poly(glycerol phosphate-N-acetyl-β-galactosaminylglycerol phosphate) with the phosphodiester bond between C-3 of glycerol and C-3 of the amino sugar. The cell walls of Nocardiopsis composta VKM Ac-2520 and N. composta VKM Ac-2521^T contain only one TA, namely 1,3-poly(glycerol phosphate) partially substituted with N-acetyl-α-glucosamine. The cell wall of Nocardiopsis metallicus VKM Ac-2522^T contains two TA. The major TA is 1,5-poly(ribitol phosphate), each ribitol unit carrying a pyruvate ketal group at positions 2 and 4. The structure of the minor TA is the same as that of N. composta. The results presented correlate well with the phylogenetic grouping of strains and confirm the species and strain specific features of cell wall TA in members of the genus Nocardiopsis.     

Cell wall teichoic acids of streptomycetes of the phenetic cluster 'Streptomyces fulvissimus'

Structures of the anionic polymers of streptomycetes Streptomyces fulvissimus VKM Ac-994(T), Streptomyces longispororuber VKM Ac-1735(T), Streptomyces aureoveticillatus VKM Ac-48(T) and Streptomyces spectabilis INA 00606 belonging to the phenetic cluster 'S. fulvissimus' were investigated by chemical and NMR spectroscopic methods. A teichoic acid from the cell wall of S. spectabilis INA 00606 was studied in more detail, and this was shown to represent 1,3-poly(glycerol phosphate) substituted with glucosamine (alpha-D-GlcNAc) and L-glutamic acid (non-stoichiometric substitution). For the first time, glutamic acid is identified as an acyl substituent in teichoic acids of streptomycetes. The polymer chain is built of the following fragments: Cell walls of other streptomycetes of the phenocluster under study contain 1,3-poly(glycerol phosphates) with glucosamine as a glycosyl substituent at O-2 of the glycerol phosphate units and L-glutamic acid and lysine as O-2 acyl substituents. Not all amino sugar residues in the polymers of these strains are N-acetylated, and the content of the glucosamine and lysine residues in the polymers of different strains is not the same. Despite certain quantitative differences in the structures of the polymers, one may consider streptomycetes of the phenocluster 'S. fulvissimus' as closely related microorganisms, the details of the structures serving as additional criteria for the determination of the species status of a strain under study.     

Teichoic acids from the cell wall of 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.     

Teichoic acids in the cell walls of Microbispora mesophila Ac-1953t and 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).     

The carbohydrate-containing cell wall polymers of some species from the cluster "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.     

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.     

A mannitol teichoic acid containing rhamnose and pyruvic acid acetal from the cell wall of Brevibacterium permense VKM Ac-2280

The cell wall of Brevibacterium permense VKM Ac-2280 contains two teichoic acids. The major polymer represents a 1,6-poly(mannitol phosphate) substituted wirh either L-rhamnose (approximately 70%, unit A) or (S)-acetal of pyruvic acid (approximately 30%, unit B) with the overall chain length approximately 10 mannitol phosphate units. [carbohydrate structure: see text] The other polymer is an unsubstituted 1,3-poly(glycerol phosphate). The structures of the polymers were established using chemical degradations and NMR spectroscopy. The data obtained may be helpful in determination of the species-specific status of newly isolated Brevibacterium strains.     

Cell wall teichoic acids of Bacillus licheniformis VKM B-511T, Bacillus pumilus VKM B-508T, and other strains previously assigned to Bacillus pumilus

The teichoic acids (TAs) of type strains, viz. Bacillus licheniformis VKM B-511^T and Bacillus pumilus VKM B-508^T, as well as phylogenetically close bacteria VKM B-424, VKM B-1554, and VKM B-711 previously assigned to Bacillus pumilus on the basis of morphological, physiological, and biochemical properties, were investigated. Three polymers were found in the cell wall of each of the 5 strains under study. Strains VKM B-508^T, VKM B-424, and VKM B-1554 contained polymers of the same core: unsubstituted 1,3-poly(glycerol phosphate) (TA I) and 1,3-poly(glycerol phosphate) with O-D-Ala and N-acetyl-??-D-glucosamine substituents (TA II and TA III??, respectively). The cell walls of two remaining strains contained TA I, TA II, and a poly(glycosylpolyol phosphate) with the following structure of repeating units: -6)-??-D-GlcpNAc(1??1)-snGro-(3-P-(TA III?) in ??Bacillus pumilus?? VKM B-711 (100% 16S rRNA gene similarity with the type strain of Bacillus safensis) and -6)-??-D-Galp-(1??2)-snGro-(3-P-(TA III?) in Bacillus licheniformis VKM B-511^T. The simultaneous presence of three different TAs in the cell walls was confirmed by the NMR spectroscopic DOSY methods. The structure of the polymers and localization of O-D-Ala residues were investigated by the chemical and NMR spectroscopic methods.     

A new neutral polymer from the cell wall of actinomycete Kineosporia aurantiaca VKM Ac-702T

The major cell wall polymer of Kineosporia aurantiaca VKM Ac-702T a representative of the suborder Frankineae, is a galactomannan with a repeating unit of the following structure: -->3)-beta-D-Galp-(1-->6)-beta-D-Manp-(1-->4)-beta-D-Manp-(1-->3)-beta-D-Galp-(1-->4)-beta-D-Manp-(1-->4)-beta-D-Manp-(1--> that has not been reported so far. This was established using chemical degradation methods and NMR spectroscopy. The polysaccharide identified in the present study provides the second example of neutral galactomannans in actinomycete cell walls. The cell wall of K. aurantiaca VKM Ac-702T also contains a minor teichoic acid, viz., 1,3-poly(glycerol phosphate) partially substituted with alpha-glucosamine residues, only part of which are N-acetylated.     

Cell wall teichoic acids ofNocardiopsis prasina VKM Ac-1880T

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

Teichoic Acids in the Cell Walls of Microbispora mesophila Ac-1953T and Thermobifida fusca Ac-1952T

The cell walls of Microbispora mesophila strain Ac-1953^T (the family Streptosporangiaceae) and Thermobifida fusca Ac-1952^T (the family Nocardiopsaceae) were found to contain teichoic acids of a poly(glycerol phosphate) nature. The teichoic acid of M. mesophila (formerly Thermomonospora mesophila) represents a 1,3-poly(glycerol phosphate) containing 5% of substituent 2-acetamido-2-deoxy--D-galactosaminyl residues. Teichoic acid of such a 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 -glucosylated 1,3-poly(glycerol phosphate).     

Anionic Polymers of the Cell Wall of <Emphasis Type="Italic">Brevibacterium linens</Emphasis> VKM Ac-2159

Unsubstituted 1,3-poly(glycerol phosphate) and two sugar-1-phosphate polymers were identified in the cell wall of Brevibacterium linens VKM Ac-2159 by NMR spectroscopy and chemical methods. A monomer of one of the sugar-1-phosphate polymers has the branched repeating unit of the following structure: -4)-[beta-D-GlcpNAc-(1-->3)]-alpha-D-Glcp-(1-P-. The repeating unit of another sugar-1-phosphate polymer has a linear structure consisting of alternating beta- and alpha-N-acetylglucosamine residues: -4)-beta-D-GlcpNAc-(1-->6)-alpha-D-GlcpNAc-(1-P-. Some part of the beta-N-acetylglucosaminyl residues bear O-ester-bound succinic acid residues at C-3. The identified sugar-1-phosphate polymers have not been described earlier in cell walls of other bacteria.     

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.     

Cell wall teichoic acids from Streptomyces daghestanicus VKM Ac-1722T and Streptomyces murinus INA-00524T

The structure of cell wall teichoic acids was studied by chemical methods and NMR spectroscopy in the type strains of two actinomycete species of the "Streptomyces griseoviridis" phenetic cluster: Streptomyces daghestanicus and Streptomyces murinus. S. daghestanicus VKM Ac-1722T contained two polymers having a 1,5-poly(ribitol phosphate) structure. In one of them, the ribitol units had alpha-rhamnopyranose and 3-O-methyl-alpha-rhamnopyranose substituents; in the other, each ribitol unit was carrying 2,4-ketal-bound pyruvic acid. Such polymers were earlier found in the cell walls of Streptomyces roseolus and Nocardiopsis albus, respectively; however, their simultaneous presence in the cell wall has never been reported. The cell wall teichoic acid of Streptomyces murinus INA-00524T was is a 1,5-poly(glucosylpolyol phosphate), whose repeating unit was [-6)-beta-D-glucopyranosyl-(1 --> 2)-glycerol phosphate-(3-P-]. Such a teichoic acid was earlier found in Spirilliplanes yamanashiensis. The 13C NMR spectrum of this polymer is presented for the first time. The results of the present investigation, together with earlier published data, show that the type strains of four species of the "Streptomyces griseoviridis" phenetic cluster differ in the composition and structure of their teichoic acids; thus, teichoic acids may serve as chemotaxonomic markers of the species.     

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-2000^T contains two types of teichoic acids: 2,3-poly(glycerol phosphate) substituted with -D-Galp at C-1 of glycerol and 1,3-poly(glycerol phosphate) substituted with -L-Rhap at OH-2 of glycerol (60%). Herbidospora cretacea VKM Ac-1997^T contains the chains of 1,3-poly(glycerol phosphate) partially substituted with -D-Galp and -D-GalpNAc at C-2 of glycerol. The majority of -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-1994^T contains poly(galactosylglycerol phosphate), with the -Galp-(12)-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.     

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.