Cell Wall Teichoic Acids: Structural Diversity, Species Speci¢City in the Genus Nocardiopsis, and Chemotaxonomic Perspective

FEMS Microbiology Reviews 25 (2001) 269^283 www.fems-microbiology.org

Cell wall teichoic acids: structural diversity, species speci¢city in the genus Nocardiopsis, and chemotaxonomic perspective

Irina B. Naumova a;*, Alexander S. Shashkov b, Elena M. Tul'skaya a, Galina M. Streshinskaya a, Yuliya I. Kozlova a, Nataliya V. Potekhina a, Lyudmila I. Evtushenko c, Erko Stackebrandt d

a School of Biology, Lomonosov Moscow State University, Moscow 119899, Russia b Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky prospect 47, Moscow 117913, Russia c VKM ^ All Russian Collection of Microorganisms, Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino 142292, Moscow Region, Russia d DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 1b, D-38124 Braunschweig, Germany

Received 11 September 2000; received in revised form 10 November 2000; accepted 15 November 2000

Abstract

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 13C 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. ß 2001 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Keywords: Teichoic acid; Taxonomy; Actinomycetes; Nocardiopsis

Contents

1. Introduction ...... 270 2. The genus Nocardiopsis ...... 271 3. Structural types of cell wall teichoic acids ...... 271 3.1. Poly(polyol phosphate) teichoic acids (type I) ...... 272 3.2. Poly(glycosylpolyol phosphate) teichoic acids (type II) ...... 273 3.3. Poly(polyol phosphate-glycosyl phosphate) teichoic acids (type III) ...... 273 3.4. Poly(polyol phosphate-glycosylpolyol phosphate) teichoic acids, mixed structure (type IV)...... 273 4. Determination of teichoic acid structures ...... 273 5. Cell wall teichoic acids of Nocardiopsis species and subspecies ...... 273 5.1. N. dassonvillei subspecies ...... 274 5.2. N. trehalosi ...... 274 5.3. N. tropica ...... 274

* Corresponding author. Fax: +7 (095) 939 43 09; E-mail: [email protected]; [email protected]

Abbreviations: GalNAc, N-acetylgalactosamine; Glc, glucose; GlcNAc, N-acetylglucosamine; Gro, glycerol; GroP, glycerol monophosphate; GroP2, glycerol diphosphate; Pyr, pyruvic acid; Rib-olP, ribitol monophosphate; Rib-olP2, ribitol diphosphate; Rib-olP3, ribitol triphosphate; Succ, succinic acid; TA, teichoic acid; tr, trace; sn, stereospeci¢c numbering

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5.4. The Nocardiopsis alba species group ...... 274 6. Application of wall teichoic acid composition in taxonomy of the genus Nocardiopsis .... 276 6.1. Structures of cell wall teichoic acids and their combinations are chemical markers of intrageneric taxa ...... 276 6.2. Di¡erentiating characteristics of Nocardiopsis species and subspecies based on the products of chemical degradation of cell wall teichoic acids ...... 277 7. 13C NMR spectra of teichoic acids are indicative of intrageneric taxa of the genus Nocar- diopsis ...... 278 7.1. 13C NMR spectra of teichoic acids of the N. dassonvillei subspecies ...... 278 7.2. 13C NMR spectrum of teichoic acid of N. trehalosi ...... 278 7.3. 13C NMR spectrum of teichoic acid of N. tropica ...... 280 7.4. 13C NMR spectra of teichoic acids of the N. alba group ...... 280 8. Conclusion ...... 280

Acknowledgements ...... 281

References ...... 281

1. Introduction more focused identi¢cation of new organisms at di¡erent taxonomic levels. The past decade has witnessed an extraordinary increase Teichoic acids are anionic carbohydrate-containing polin methodological approaches towards bacterial system- ymers, which occur in many Gram-positive bacteria and atics, including the introduction of rapid sequence proto- are associated with the cytoplasmic membrane (lipotei- cols for the analysis of RNA and DNA allowing the de- choic acids) [10] or covalently linked by phosphodiester termination of relationships between prokaryotic bridges to muramic acid residues in peptidoglycan (cell organisms. However, due to di¡erences in the tempo and wall teichoic acids) [11]. The possible application of lipo- mode of gene and protein evolution that exist between teichoic acids in bacterial taxonomy has been discussed organisms, the phylogenetic branching pattern of bacterial previously [10]. The data on the cell wall teichoic acids lineages per se cannot be used to de¢ne hierarchical clas- accumulated by now for Gram-positive bacteria, mainly si¢cations. Additional information is needed to facilitate actinomycetes, have demonstrated their potential for bac- the delineation of genera, species and higher taxa at the terial systematics [12^15]. phenetic level and to meet the demands of both phyloge- The physiological role of teichoic acids is thought to be netic coherence and the requirement for common pheno- in ion exchange and control of the activity of autolytic typic characters necessary for unambiguous description of enzymes that, in turn, are important for growth and divi- bacterial taxa and reliable identi¢cation of their members. sion of the bacterial cell [16]. Teichoic acids may constitute Large molecules and polymers of cells and cell enve- 60% of the cell wall and are essential for the normal func- lopes have contributed signi¢cantly to an improved classi- tioning of the bacterial cell, which spends a signi¢cant ¢cation of the order Actinomycetales embracing a large amount of energy on the synthesis of these polymers. group of Gram-positive bacteria of immense physiological, About 10 genes were shown to be involved in the control biochemical and morphological diversity, which are well of the synthesis of glycosylated poly(glycerol phosphate) known for their capacity to produce antibiotics and other teichoic acid in Bacillus sp. and of an oligomeric unit link- biologically active substances [1]. However, the limited ing this polymer to peptidoglycan [17^19]. The cell dis- number of the traditionally used chemotaxonomic charac- plays a morphological abnormality when the content of teristics based on these cell constituents, i.e., cell wall che- this polymer in the cell wall is below the norm, and mu- motype [2,3], phospholipid type [4], menaquinone [5] and tant bacilli lacking teichoic acids are not found, since they fatty acid composition [6], presence and type of mycolic appear to be unviable [16]. Other functions of these poly- acids [7], often does not provide su¤cient markers for mers are related to phage binding [20] and immunogenicity phenetic delineation of the actinomycete taxa newly de- [21]. scribed or revised mainly on the basis of phylogenetic The involvement of cell wall teichoic acids in vital cell data. There is also a special problem of reliable description functions and their great structural diversity (see below), and di¡erentiation of bacterial species in accordance with expressed by the information coded in various genes, as the current bacterial species de¢nition [8]. On the other well as their wide occurrence in Gram-positives, suppose a hand, many cell polymers and molecules have still not correlation of these polymers with other taxonomic prop- been determined or evaluated for their taxonomic value erties and suggest their applicability to the systematics of [9]. Their characterization from the taxonomic standpoint the respective microorganisms. could provide new tools for bacterial classi¢cation and a The occurrence of cell wall teichoic acids cannot be

FEMSRE 713 1-5-01 I.B. Naumova et al. / FEMS Microbiology Reviews 25 (2001) 269^283 271 predicted from the phylogenetic position of taxa but, if hydrolysates). Subsequently, the genus description was present, they identify phylogenetically related organisms, amended by Grund and Kroppenstedt [36], and only the i.e., all tested strains belonging to the suborder Strepto- organisms with the cell wall chemotype III C, phospholip- sporanginea (including the families Streptosporangiaceae, id type P III (phosphatidylcholine and phosphatidyl-meth- Nocardiopsaceae, and Thermomonosporacea) and most ylethanolamine as characteristic phospholipids), menaqui- organisms of the family Streptomycetaceae [15,22]. The none MK-10 with variable degrees of saturation as the absence of teichoic acids in some phylogenetically de¢ned major isoprenoid quinones, fatty acids of the 3d type, taxa is worth mentioning in the phenotypic description of and a DNA G+C content ranging between 64 and 71 the respective taxa, such as in the suborder Pseudonocar- mol% were proposed to be included in this genus. The dineae, and in the members of the genera Cellulomonas, taxonomic coherence of this generic grouping was con- Promicromonospora, Micrococcus, Koccuria, Kytococcus, ¢rmed by 16S rDNA sequence analysis [40,41]. Nesterenkonia, and Dermabacter [23,24], which are char- Members of the genus Nocardiopsis are known mostly acterized by peptidoglycan containing lysine. The structur- as soil and clinical isolates [35,36,42^50]. A number of al elements of cell wall teichoic acids are re£ected in some strains were also obtained from an Antarctic glacier, widely used chemotaxonomic characteristics of the generic dust, the rhizosphere of actinorhizal plants, the gut tract and suprageneric levels, e.g., madurose (3-O-methyl-D-ga- of animals, saltern, etc. [40,45,51^53]. Like other actino- lactose) [15,25] which is the diagnostic component of the mycetes, Nocardiopsis spp. are capable of producing me- cell wall pattern B [2,3]. tabolites with biological activities [45,50,54,55]. It should An observation that the species within a genus may be also mentioned that a number of organisms cited as have structurally dissimilar teichoic acids was reported Nocardiopsis spp. in some previous publications were re- for the ¢rst time for staphylococci [26]. The relevant classi¢ed into other genera [45,56^59]. data published for some actinomycete genera (e.g., Strep- The current genus Nocardiopsis contains 10 validly de- tomyces, Actinomadura, Glycomyces, Nocardioides) have scribed species and eight of them, with the exception of N. demonstrated that the structures of the polymers can be halophila and N. kunsanensis, are veri¢ed by DNA^DNA indicative of the species [15,25,27^34]. relatedness data [37^40,42,53]. These two species, however, However, the taxonomic value of the teichoic acid struc- were shown to represent distinct phyletic lines that can be tures was not estimated for the majority of actinomycete equated with genomic species [53]. At the phenetic level, groups, including the large genus Streptomyces, because the di¡erentiation of Nocardiopsis species is based on the the data on these polymers are insu¤cient and the validity color of the aerial and vegetative mycelium; di¡usible pig- of most species has not yet been veri¢ed phylogenetically, ments, menaquinone composition, and physiological char- in accordance with the current bacterial species de¢nition acteristics, including halotolerance, growth at di¡erent pH [8]. The genus Nocardiopsis [35,36] is an example where and temperatures [36,38^40,42,45,53]. Recent study of the most species have been justi¢ed by DNA^DNA reassoci- cell wall teichoic acids shows that the structure and com- ation data [37^40] and the structures and composition of position of these polymers can be used for di¡erentiation teichoic acids have been identi¢ed in the respective organ- and identi¢cation of Nocardiopsis species as well. isms. In this work, the main structural types and subtypes of the cell wall teichoic acid structures found in bacteria to 3. Structural types of cell wall teichoic acids date are considered and the data on the cell wall teichoic acids of the genus Nocardiopsis are summarized from the Among the cell wall teichoic acids studied, the following taxonomic standpoint. This genus should serve as an ex- four structural types can be determined depending on the ample for the usefulness of teichoic acids in the polyphasic composition of main chains: polymers with the chain con- approach to actinomycete classi¢cation. The material pre- sisting of only polyol phosphate residues or glycosylpolyol sented illustrates that structures, products of chemical deg- phosphate residues, joined by phosphodiester bonds (types radation of the polymers, and NMR spectra are indicative I and II, respectively), and polymers of mixed structures, of species and subspecies of the genus Nocardiopsis, and glycosyl-1-phosphate in addition to polyol phosphate res- these characteristics can be fruitfully used in taxonomic idues (type III), or alternating units of poly(polyol phos- practice. phate) and poly(glycosylpolyol phosphate) (type IV):

2. The genus Nocardiopsis

The genus Nocardiopsis was described by J. Meyer [35] to comprise actinomycetes with fragmenting mycelium and cell wall chemotype III C (with the meso isomer of diaminopimelic acid and no characteristic sugars in whole-cell

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Several factors contribute to the structural diversity of erythritol, or arabitol as a polyol in the main chain, and, teichoic acids: the di¡erent nature of the polyol and sugar respectively, the main chain is poly(glycerol phosphate), (amino sugar) residues that might serve both as an integral poly(ribitol phosphate), poly(mannitol phosphate), poly- part of the main chain and as lateral branches linked to (erythritol phosphate) or poly(arabitol phosphate) chain the polyol residues via glycosyl bonds; di¡erent kinds of (Table 1). Eight subtypes can be distinguished within phosphodiester bonds; varying O-acyl residues. Chains this type, based on variations of the polyols and the local- void of glycosyl and acyl substituents are known. Below, ization of phosphodiester bonds. these main types of teichoic acids and their occurrences Poly(glycerol phosphates) are represented by 1,3-poly- are brie£y considered. (glycerol phosphate) and 2,3-poly(glycerol phosphate) polymers (Table 1). In 1,3-poly(glycerol phosphates), phos- 3.1. Poly(polyol phosphate) teichoic acids (type I) phodiester bonds link C-1 and C-3 of glycerol residues (subtype I-G 1,3) [11,13,19,27,32,60^67] and in 2,3-poly- Polymers of type I contain glycerol, ribitol, mannitol, (glycerol phosphates), C-2 and C-3 residues (subtype I-G

Table 1 Types and subtypes of teichoic acids Subtype Polyol Presence of Position of carbons in polyol or Position of glycosyl Occurrence of teichoic acids of main sugar in main sugar, linked through their OH substituent at C of in bacterial genera chain chain groups by phosphodiester bonds polyola;b Type I, poly(polyol phosphates) I-G 1,3 Glycerol Glycerol, C-1 and C-3 C-2 Arthrobacter [65], Bacillus [11,61], Brevibacterium [13,67], Glycomyces [32], Herbidospora [93], Lactobacillus [11,61], Microbisporad, Nocardiopsis [19,60,74,75], Nonomuraea [63], Planotetraspora [93], Spirilliplanes [93], Staphylococcus [11,61], Streptomyces [19,27,62,64,66], Thermobi¢dad I-G 2,3 Glycerol Glycerol, C-2 and C-3 C-1 Bacillus [68], Glycomyces [32], Planotetraspora [93], Streptomyces [19,27,69] I-R 1,5 Ribitol Ribitol, C-1 and C-5 C-4 Bacillus [11,61], Brevibacterium [13], Lactobacillus [11,61], Listeria [14,70], Nocardioides [34], Nocardiopsis [60,71,74], Staphylococcus [11,61], Streptomyces [15,19,72,73] I-R 3,5 Ribitol Ribitol, C-3 and C-5 No Nocardiopsis [74,75] I-E 1,4 Erythritol Erythritol, C-1 and C-4 C-2 Brachybacterium [76,77], Glycomyces [31] I-M 1,6 Mannitol Mannitol, C-1 and C-6 C-2 Brevibacterium [13,67,78] I-M 4,6 Mannitol Mannitol, C-4 and C-6 No Bi¢dobacterium [79] I-Ac Arabitol Unknown C-4 Agromyces [80]

Type II, poly(glycosylpolyol phosphates) II-GS 3,3 Glycerol Sugar Glycerol, C-3; sugar, C-3 C-1 Actinomadura [29], Bacillus [86], Nocardioides [33] II-GS 3,4 Glycerol Sugar Glycerol, C-3; sugar, C-4 C-1 Streptomyces [85] II-GS 3,6 Glycerol Sugar Glycerol, C-3; sugar, C-6 C-1 or C-2 Actinocorallia [93], Actinomadura [25,30], Actinoplanes [19,84], Bacillus [68,82,83], Lactobacillus [11], Spirilliplanes [93], Streptomyces [15,84] II-RS 5,3 Ribitol Sugar Ribitol, C-5; sugar, C-3 C-1 Agromyces [87] II-RS 5,6 Ribitol Sugar Ribitol, C-5; sugar, C-6 C-1 Streptococcus [88,89]

Type III, poly(polyol phosphate-glycosyl phosphates), mixed structures III-GS Glycerol Sugar Glycerol, C-1 and C-3; No or C-2 Staphylococcus [61,90] sugar, C-1 and C-6

Type IV, poly(polyol phosphate-glycosylpolyol phosphates), mixed structures IV-GS Glycerol Sugar Glycerol, C-1 and C-3; C-2 Nocardiopsis [91,92] glycerol, C-3 and sugar, C-3 or C-4 aThe most common glycosyl substituents are: glucose, galactose, rhamnose and N-acetylated amino sugars. b O-D-Alanyl, O-L-lysyl, O-acetyl, O-succinyl or pyruvate ketal groups can sometimes be substituents at C of polyols. cThe position of the phosphodiester bond is unknown. dN.V. Potekhina, A.S. Shashkov, L.I. Evtushenko and I.B. Naumova, in preparation.

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2,3) [19,27,32,68,69]. Glucose, galactose, and N-acetylated 3.4. Poly(polyol phosphate-glycosylpolyol phosphate) amino sugars are the most common glycosyl substituents teichoic acids, mixed structure (type IV) which are located at C-1 or C-2 of glycerol. O-D-Alanyl, O-L-lysyl or O-acetyl are also found [19]. Poly(glycerol The core of this polymer type consists of alternating phosphates) are the most widely occurring bacterial cell units of poly(glycerol phosphate) and poly(glycosylglycer- wall teichoic acids. ol phosphate) chains. Both glycerol phosphate and L-N- Among poly(ribitol phosphates), the subtype I-R 1,5 pol- acetylgalactosaminylglycerol phosphate residues are ymers (phosphodiester bonds link C-1 and C-5 of ribitol) present within the integral portion of the chain. This struc- are the most frequently found and occur in many Gram- ture has been found exclusively in N. dassonvillei subspe- positives [11,13^15,19,34,60,61,70^74]. The following sub- cies [91,92] and is discussed in detail in Section 5.1. stituents were determined in these polymers: glucose, rhamnose, 3-O-methylrhamnose, N-acetylated amino sugars, both ribitol phosphate and glycerol phosphate side 4. Determination of teichoic acid structures chains, as well as O-L-lysyl, O-D-alanyl, and O-succinyl residues and pyruvate ketal groups [19]. Only the unsub- Cells for teichoic acid analysis may be grown on any stituted 3,5-poly(ribitol phosphate) teichoic acid (subtype suitable liquid medium supplemented with K2HPO4 in- I-R 3,5) has been found to date (in Nocardiopsis alba and volved in the polymer biosynthesis. Usually cells grown Nocardiopsis prasina) [74,75]. in peptone-yeast extract medium (5 g peptone, 3 g yeast

The details of structures of the type I teichoic acids with extract, 5 g glucose and 0.2 g K2HPO4 per liter, pH 7.0) other polyols in the main chain are given in Table 1. These are used [28]. Methods of extraction, puri¢cation and are poly(erythritol phosphate) found in Glycomyces tenuis identi¢cation of cell wall teichoic acids of actinomycetes [31] and Brachybacterium spp. [76,77], poly(mannitol phos- have been described in detail previously [62,91]. For the phate) detected in Brevibacterium spp. [13,67,78] and Bi¢- cell wall teichoic acid isolation, a native cell wall was ob- dobacterium bi¢dum [79], and poly(arabitol phosphate) oc- tained from crude mycelium by fractional centrifugation curring exclusively in Agromyces cerinus subsp. cerinus after preliminary disruption by sonication, and puri¢ed [80]. using 2% sodium dodecyl sulfate to avoid possible con- tamination with membrane compounds, including lipotei- 3.2. Poly(glycosylpolyol phosphate) teichoic acids choic acids. The polymers were extracted with 10% (type II) trichloroacetic acid, puri¢ed by ion exchange chromatog- raphy and identi¢ed using chemical and NMR spectrosco- Five subtypes of poly(glycosylpolyol phosphate) poly- py methods. The chemical methods consist of the analysis mers can be de¢ned, based on the combination of certain of products formed on acid and alkaline degradation of polyols (glycerol or ribitol) and sugars in the main chain the polymers. The molecular mass of the extracted teichoic and the position of phosphodiester bonds between the acids was determined on Sephadex G-50 by comparison polyol and sugar (Table 1). In the known poly(glycosyl- with teichoic acids of known molecular mass [62]. NMR glycerol phosphates), an sn-glycerol-3-phosphate residue is has been used to analyze the 13C and 1H NMR spectra of always involved in the phosphodiester bond [61,81] and the teichoic acids, which provided information on the de- the glycosyl substituents are usually positioned at C-1 of gree of polymer regularity, the type of alditol and sugars, glycerol of the main chain. The polymers with the phos- the number of sugar residues in the repeating unit, their phodiester bond located at C-6 of the sugar substituent ring sizes, the non-carbohydrate substituents, etc. In most (subtype II-GS 3,6) are the most widespread [11,15,19, cases, a combination of chemical and NMR techniques is 25,30,68,82^84]. Other structural variations are character- necessary. For example, chemical degradation data alone ized by the phosphodiester bonds at C-3 or C-4 of the are inadequate for identifying the bonds between the re- sugar (subtypes II-GS 3,3 and II-GS 3,4) [29,33,85,86]. peating units of the teichoic acids of Nocardiopsis dasson- Poly(glycosylribitol phosphates) (subtypes II-RS 5,3 and villei, but this can be done successfully by NMR spectros- II-RS 5,6) are known to contain the glycosyl substituents copy. On the other hand, the chemical approaches permit at C-1 and are found in Agromyces cerinus subsp. nitratus one to identify minor components of the polymers which [87] and streptococci [88,89]. cannot be detected by NMR.

3.3. Poly(polyol phosphate-glycosyl phosphate) teichoic acids (type III) 5. Cell wall teichoic acids of Nocardiopsis species and subspecies A teichoic acid of this type, containing glycerol phosphate and sugar-1-phosphate within the integral portion of Until now, teichoic acid structures have been studied in the chain (Table 1), has been identi¢ed only in staphylo- 27 strains belonging to seven validly described Nocardiop- cocci [61,90]. sis species veri¢ed by data on DNA^DNA relatedness

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[37^40]. These polymer structures, their indicative features ol, glucose, and inorganic phosphate are formed. The and important products of their chemical degradation are polymer chain consists of about 31^33 repeating units summarized in this section. [60].

5.1. N. dassonvillei subspecies 5.3. N. tropica

5.1.1. N. dassonvillei subsp. dassonvillei The cell wall of N. tropica VKM Ac-1457T contains All 18 strains of N. dassonvillei subsp. dassonvillei nearly 45% of the teichoic acid represented by an assort- studied [40], including the type strain VKM Ac-797T, ment of heterogeneous 1,5-poly(ribitol phosphate) chains were found to contain in their cell walls about 20% tei- (subtype I-R 1,5) with lateral glycerol phosphate oligo- choic acid of identical structure and of the subtype IV-GS. mers. The latter are bound to C-3 of the portion of ribitol The core is composed of alternating units of poly(glycerol residues of the main chains (Fig. 1d). The chains di¡er in phosphate) and poly(glycosylglycerol phosphate) chains the number of glycerol phosphate units in the lateral olig- and has about 13 repeating units which consist of both omers and the number of substituted ribitol phosphate glycerol phosphate and L-N-acetylgalactosaminylglycerol units [71]. The polymer yields identical products on acid phosphate residues (Fig. 1a). The polymer of this structure and alkaline hydrolyses: glycerol mono- and diphos- gives unusual products on alkaline hydrolysis, including phates, glycerol, ribitol mono-, di-, and triphosphates, an- phosphodiester E-3, characteristic of N. dassonvillei subsp. hydroribitol phosphate, ribitol, anhydroribitol, and inor- dassonvillei, which contains two glycerol residues, one N- ganic phosphate. The molecular mass of the chains is 5.4^ acetylgalactosaminyl residue, and three phosphate groups 9 kDa. [P-(3/4)-GalNAc-1C2-Gro-3-P-1-Gro-2-P]. On acid hydrolysis, glycerol mono- and diphosphates, galactosamine, 5.4. The Nocardiopsis alba species group glycerol, and inorganic phosphate are formed [91]. The organisms of four species of this group are charac- 5.1.2. N. dassonvillei subsp. albirubida terized by di¡erent combinations of structurally distinct The cell wall of N. dassonvillei subsp. albirubida VKM sets of the following polymers: unsubstituted 3,5-poly(ri- Ac-1882T contains about 18% of the teichoic acid which bitol phosphates) (TA1, subtype I-R 3,5), 1,3-poly(glycerol resembles in its structure the polymers of N. dassonvillei phosphates) substituted with K-N-acetylglucosamine at C- subsp. dassonvillei and belongs to the same structural IV- 2 of glycerol residues (TA2, subtype I-G 1,3), and 1,5- GS subtype. However, the phosphodiester bond is posi- poly(ribitol phosphates) with 2,4-pyruvate ketal groups tioned at C-4 of the amino sugar rather than at the C-3 (TA3, subtype I-R 1,5) [60,74,75]. The structurally analo- characteristic of N. dassonvillei subsp. dassonvillei, and the gous teichoic acids of di¡erent species vary in molecular C-3 position is occupied by an O-succinyl substituent in mass and, accordingly, in chain length. In addition, the most glycosyl residues (Fig. 1b) [92]. These di¡erences re- TA2 polymers di¡er in the proportion of substituted glyc- sult in a di¡erent behavior of the polymer on alkaline erol phosphate units. hydrolysis. In contrast to N. dassonvillei subsp. dassonvil- Both acid and alkaline hydrolysis of the TA1 polymer lei, only trace amounts of ester E-3 are formed from the yield identical phosphoric esters, ribitol mono- and di- polymer of N. dassonvillei subsp. albirubida. The molecular phosphates. Formaldehyde and phosphorus in equimolar mass of the teichoic acid is close to 10 kDa, which is amounts as the products of periodate oxidation of the consistent with the presence of 14^15 repeating units. polymer di¡erentiate this polymer from unsubstituted 1,5-poly(ribitol phosphate). On acid hydrolysis of TA2, 5.2. N. trehalosi glycerol mono- and diphosphates, glucosamine, glycerol, and inorganic phosphate are formed. Alkaline degradation The cell wall of N. trehalosi VKM Ac-942T has about yields the same phosphoric esters, glycerol, and a small 30% of the 1,3-poly(glycerol phosphate) (subtype I-G 1,3) amount of glycerol phosphodiesters containing N-acetyl- substituted with L-glucosyl residues (Fig. 1c). The degree glucosamine. TA3 is resistant both to alkaline hydrolysis of substitution is about 60%. The characteristic feature and to periodate oxidation. Ribitol mono- and diphos- of this polymer is resistance to alkaline degradation. On phates, ribitol, inorganic phosphate, and pyruvic acid are acid hydrolysis, glycerol mono- and diphosphates, glycer- the major products of acid hydrolysis.

C Fig. 1. The repeating units of teichoic acids of the Nocardiopsis species and subspecies. a: Poly(glycerol phosphate-L-N-acetylgalactosaminylglycerol phosphate) (N. dassonvillei subsp. dassonvillei). b: Poly(glycerol phosphate-L-N-acetylgalactosaminylglycerol phosphate) with O-succinyl residues (N. dassonvillei subsp. albirubida). c: 1,3-Poly(glycerol phosphate) with L-glucose (N. trehalosi). d: 1,5-Poly(ribitol phosphate) with side chains of glycerol phosphate oligomers (N. tropica). e: Unsubstituted 3,5-poly(ribitol phosphate), TA1 (N. alba, N. prasina). f: 1,3-Poly(glycerol phosphate) with K-N-acetylglucosamine, TA2 (N. alba, N. listeri, N. prasina). g: 1,5-Poly(ribitol phosphate) with 2,4-pyruvate ketal group, TA3 (N. alba, N. listeri, N. lucentensis).

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5.4.1.. N. alba poly(ribitol phosphate) with each ribitol phosphate unit The cell walls of three strains of N. alba (VKM Ac- substituted by a 2,4-pyruvate ketal group; its molecular 1883T, VKM Ac-1879, VKM Ac-1884) were found to con- mass was 10 kDa (equivalent to approximately 36^37 ri- tain 3^3.5% of phosphorus and all the above polymers, bitol phosphate units) [60]. TA1, TA2, and TA3, in varying amounts [74]. TA1 (unsubstituted 3,5-poly(ribitol phosphate)) of this species con- 5.4.4. N. lucentensis sisted of 26^27 ribitol phosphate units (Fig. 1e). TA2 of N. Strain VKM Ac-1962T possessed only the TA2 polymer alba is a 1,3-poly(glycerol phosphate) with about 10% having a substitution with K-N-acetylglucosamine residues glycerol phosphate units substituted with K-N-acetylglu- close to 50%, which is similar to the analogous polymer of cosamine (Fig. 1f); the polymer chain has about 40 glyc- N. listeri. The cell wall contained about 30% teichoic acid. erol phosphate units. TA3 is a 1,5-poly(ribitol phosphate) The molecular mass has been approximated to 8 kDa with each ribitol phosphate unit carrying a 2,4-pyruvate corresponding to 31^32 glycerol phosphate residues [60]. ketal group (Fig. 1g). The polymer chain consists of about 18 ribitol phosphate units. 6. Application of wall teichoic acid composition in 5.4.2. N. prasina taxonomy of the genus Nocardiopsis The type strain N. prasina VKM Ac-1880T contains two teichoic acids, TA1 with a chain length close to that of the 6.1. Structures of cell wall teichoic acids and their analogous polymer of N. alba, and TA2, where 10% glyc- combinations are chemical markers of intrageneric taxa erol phosphate units were substituted by the residues of K- N-acetylglucosamine and 5% units had O-acetyl groups. As shown above and summarized in Table 2, the No- The molecular mass of the N. prasina TA2 is close to cardiopsis species and subspecies are de¢nable by the com- 6 kDa [75]. position of teichoic acids, and these polymers can be considered a valuable chemotaxonomic marker for the 5.4.3. N. listeri intrageneric taxa of Nocardiopsis. The full identi¢cation In the cell wall of N. listeri VKM Ac-1881T, TA2 and of the polymers by chemical and NMR methods, as de- TA3 were determined. About 50% of poly(glycerol phos- scribed in detail elsewhere [60,71,74,75,91,92], permits one phate) residues in TA2 were substituted with K-N-acetyl- to determine the species and subspecies a¤liations of glucosamine (in comparison with 10% substitution in the strains assigned to the genus by the common phenetic equivalent polymer of N. alba). The molecular mass of the generic criteria or by 16S rDNA analysis [36,40,41]. polymer was 6.6 kDa, which corresponds to approxi- More importantly, some cell wall teichoic acids have never mately 22^23 glycerol phosphate residues. TA3 was 1,5- been reported in other prokaryotes and have been found

Table 2 Teichoic acids of Nocardiopsis species and subspecies Teichoic acid N. dassonvillei N. dassonvillei N. N. N. N. N. N. Typea Sub- subsp. dassonvillei subsp. albirubida trehalosi tropica alba listeri prasina lucentensis typea Poly(glycerol phosphate-L-N- + 3 3 33333 IV IV-GS acetylgalactosaminylglycerol phosphate) Poly(glycerol phosphate-L-N- 3 + 3 33333 IV IV-GS acetylgalactosaminylglycerol phosphate) with O-succinyl residues 1,3-Poly(glycerol phosphate) 33+ 33333 I I-G 1,3 with L-glucose 1,5-Poly(ribitol phosphate) 333+ 3333 I I-R 1,5 with side chains of glycerol phosphate oligomers Unsubstituted 3,5-poly(ribitol 3333+ 3 + 3 I I-R 3,5 phosphate) (TA1) 1,3-Poly(glycerol phosphate) 3333+ + + + I I-G 1,3 with K-N-acetylglucosamine (TA2) 1,5-Poly(ribitol phosphate) 333++333 I I-R 1,5 with 2,4-pyruvate ketal group (TA3) aSee Table 1.

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Table 3 Di¡erentiating characteristics of Nocardiopsis species and subspecies based on products of acid and alkaline degradation of common fractions of cell wall teichoic acidsa Characteristic N. dassonvillei N. dassonvillei N. N. N. N. N. N. subsp. dassonvillei subsp. albirubida trehalosi tropica alba listeri prasina lucentensis Products of acid degradation Glycerol + + + + + + + Ribitol 333+++33 Glucose 33+ 33333 Glucosamine 3333++++ Galactosamine + + 333333 Pyruvate 3333++33 Succinate 3 + 333333

Products of alkali degradation Glycerol tr tr 3 +++++

GroP, GroP2 ++3 +++++ Ribitol 333+trtrtr3

Rib-olP, Rib-olP2 333++3 + 3 Rib-olP3 333+ 3333 E-3b + 3 333333 Gro-phosphodiester 3333++++ a+, positive; 3, negative; tr, trace. bPhosphodiester containing two glycerol residues, one N-acetylgalactosamine, and three phosphate groups: P-(3/4)-GalNAc-1C2-Gro-3-P-1-Gro-2-P [91]. only in certain species of the genus (Table 2): the unsub- additional valuable argument for elevation of the strain N. stituted 3,5-poly(ribitol phosphate) (TA1), pyruvilated 1,5- dassonvillei VKM Ac-1882T to separate subspecies level poly(ribitol phosphate) (TA3), 1,5-poly(ribitol phosphate) and reclassi¢cation as N. dassonvillei subsp. albirubida [40]. with the lateral glycerol phosphate oligomers, an unusual teichoic acid with the core composed of the alternating 6.2. Di¡erentiating characteristics of Nocardiopsis species units of poly(glycerol phosphate) and poly(glycosylglycer- and subspecies based on the products of chemical ol phosphate) chains in N. dassonvillei subsp. dassonvillei, degradation of cell wall teichoic acids and a similar polymer with glycosyl residues occupied by the O-succinyl residues. Along with the fully identi¢ed structures of cell wall The structures and combinations of teichoic acids in the teichoic acids, the main products of acid and alkali deg- Nocardiopsis spp. under discussion correlated well with the radation of the common fraction of the polymers in each phylogenetic grouping of strains, determined by 16S Nocardiopsis species and subspecies can be used to identify rDNA sequence analysis [40,41]. The organisms of N. das- Nocardiopsis strains at the intrageneric level. As shown in sonvillei with the poly(polyol phosphate-glycosylpolyol Table 3, the species and subspecies of the genus di¡er by phosphate) teichoic acids fall into a common phylogenetic acid hydrolysate patterns composed of various combina- cluster. The members of the Nocardiopsis alba group (N. tions of glycerol, ribitol, glucose, glucosamine, galactos- alba, N. prasina, N. listeri, and N. lucentensis) containing amine, pyruvate, and succinate. All these compounds the di¡erent combinations of TA1, TA2 and TA3 formed can easily be detected using chromatographic methods. the second close phylogenetic cluster. The species N. tro- Determination of some diagnostic products of alkaline pica and N. trehalosi, which possess markedly di¡erent hydrolysates of whole fractions of the polymers (glycerol, teichoic acids (1,5-poly(ribitol phosphate) with lateral ribitol, their mono-, di-, triphosphates and two di¡erent glycerol phosphate oligomers and 1,3-poly(glycerol phos- phosphodiesters) increases the taxonomic resolution of phate) substituted with L-glucosyl residues), show a lower this approach and reliability of identi¢cation of the poly- phylogenetic relationship with both N. dassonvillei subspe- mers and the respective taxa. For instance, N. alba and N. cies and the N. alba species group. listeri, which are characterized by similar pro¢les of acid It is also interesting that the presence of succinic acid degradation products, can be di¡erentiated from each oth- with a free carboxyl group in the teichoic acid of N. das- er by mono- and diphosphates of ribitol in the products of sonvillei subsp. albirubida, in contrast to N. dassonvillei alkaline hydrolysis. While galactosamine in acid hydroly- subsp. dassonvillei, should make its cell wall more anionic sates of teichoic acids is characteristic of both N. dasson- than that of the latter. This fact undoubtedly in£uences villei subspecies, the phosphorus ester E-3 (consisting of the cell wall peculiarities and functions, which are associ- two glycerol residues, one N-acetylgalactosaminyl residue, ated with ion exchange, control of the activity of autolytic and three phosphate groups) in alkaline hydrolysates of enzymes, bacteriophage recognition, etc. This served as an the polymers is a reliable marker of N. dassonvillei subsp.

FEMSRE 713 1-5-01 278 I.B. Naumova et al. / FEMS Microbiology Reviews 25 (2001) 269^283 dassonvillei, the most widely occurring member of the ge- dassonvillei, alternating units of poly(glycerol phosphates) nus. The species N. trehalosi is well recognizable by the and poly(glycosylglycerol phosphates), is typical of a reg- presence of glucose in acid hydrolysate. ular polymer, containing signals with the same integral Some diagnostic components of teichoic acid can be intensities (Fig. 2a, Table 5). Some signals were broadened detected in acid hydrolysates of whole cell walls without or split into doublets or triplets owing to the interaction the extraction of pure polymers. This approach gives a with 31P nuclei. The total number of signals in the spec- simple key for identi¢cation of the members of the genus trum was 14 (taking into account the multiple intensity of Nocardiopsis in microbiological practice. The products of some of them, e.g., at 67.3 ppm). Acetamidopyranose with acid hydrolysates of whole cell walls, i.e., glycerol, ribitol, a L-con¢guration of the glycoside link was identi¢ed in the succinate, or pyruvate, combined with the menaquinone polymer by signals in the characteristic regions of the composition and growth temperatures (Table 4), are ex- spectrum: a peak in the resonance region of anomeric tremely helpful characteristics for identi¢cation of the No- carbons with a chemical shift of 102.15 ppm and a peak cardiopsis species and subspecies discussed. Three other in the resonance region of nitrogen-bound carbons at 52.5 validly described species of the genus, N. halophila, N. ppm, as well as the signals from NHCOCH3 at 176.1 and kunsanensis and N. synnemataformans, in which teichoic 23.7 ppm. The spectrum also contains signals typical of acids have not yet been studied, are clearly distinguished 1,3-poly(glycerol phosphate) (67.3 ppm, CH2 and 70.65 from other species by their resistance to high concentra- ppm, CH) [91]. tions of salt, menaquinone composition, and morpholog- The 13C NMR spectrum of teichoic acids of N. dasson- ical features [39,42,53]. villei subsp. albirubida is complex due to the presence of two sets of chemical shifts of carbon atoms: one arises from the repeating units carrying the O-succinyl residue, 7. 13C NMR spectra of teichoic acids are indicative of the other from the repeating units free of the acid residue intrageneric taxa of the genus Nocardiopsis (Fig. 2b, Table 5). The peculiarity of the 13C NMR spectrum of this subspecies is the presence of signals of suc- Comparison of 13C NMR spectra of teichoic acids com- cinic acid at 31.9 and 32.7 ppm (C-2 and C-3, respec- posing the cell walls of Nocardiopsis species and subspecies tively). The non-stoichiometric amount of this shows that they are signi¢cantly di¡erent (Fig. 2, Table 5), substituent provides a spectrum typical of polymers with and therefore the respective polymers are recognizable a masked regularity of the structure [92]. without chemical analyses. This section demonstrates the individualities of polymer 13C NMR spectra of Nocardiop- 7.2. 13C NMR spectrum of teichoic acid of N. trehalosi sis spp. and o¡ers another way to identify the organisms of the genus based on their cell wall teichoic acid compo- The 13C NMR spectrum of teichoic acids of N. trehalosi sition. was typical of 1,3-poly(glycerol phosphate) partially substituted with a sugar [60]. The signals at 67.4^67.7 ppm are 7.1. 13C NMR spectra of teichoic acids of the characteristic of 1,3-poly(glycerol phosphate) chains. N. dassonvillei subspecies However, the signals are shifted to a higher region of the spectrum when glycerol residues are substituted in po- The spectrum of teichoic acids of N. dassonvillei subsp. sition 2 by sugar residues (66.4^66.0 ppm). There are typ-

Table 4 Di¡erentiating characteristics of Nocardiopsis species and subspecies based on whole cell wall compounds (acid hydrolysis), growth temperature and menaquinone composition Characteristic N. dassonvillei N. dassonvillei N. trehalosi N. tropica N. alba N. listeri N. prasina N. lucentensis subsp. dassonvillei subsp. albirubida Products of acid degradation of whole cell wall Glycerol +a + ++++++ Ribitol 333++++3 Glucose +b 3 + 33333 Pyruvate 3333++33 Succinate 3 + 333333

Other characteristicsc Growth at 45³C 33+ 33333

Major menaquinone MK-10H2 to MK- MK-10 MK-10H4, MK-10H6, MK-10H4, MK-10, MK- MK-10H4, MK-10H6, 10H6 MK-10H6 MK-10H8 MK-10H6 10H2 MK-10H6 MK-10H8 a+, positive; 3, negative. bCompound originated not from cell wall teichoic acid. cData from [36,38^40].

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Table 5 13C NMR spectral data on cell wall teichoic acids of Nocardiopsis species

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7.4. 13C NMR spectra of teichoic acids of the N. alba group

The 13C NMR spectrum of 3,5-poly(ribitol phosphate) (TA1) of N. alba and N. prasina contains ¢ve main signals; some of them have a characteristic splitting at phosphorus atoms. The signal at 78.6 ppm is typical of ^ CHOP^ groups, while the signal at 68.3 ppm is character-

istic of ^CH2OP^ groups (Fig. 2e, Table 5) [74,75]. The 13C NMR spectra of TA2 (N. alba, N. prasina, N. listeri, and N. lucentensis) are typical of 1,3-poly(glycerol phosphate) partially substituted with a sugar. The signals at 67.4^67.7 ppm are characteristic of 1,3-poly(glycerol phosphate) chains. The signals are shifted to a higher region of the spectrum when glycerol residues are substituted in position 2 by sugar residues (66.4^65.7 ppm). There is a typical signal in the resonance region of anomeric carbons at 98.0 ppm and the signal of carbon bearing nitrogen at 55.0 ppm for K-D-N-acetylglucosamine (Fig. 2f, Table 5) [74,75]. The spectrum of the pyruvilated 1,5-poly(ribitol phosphate) (TA3) of N. alba and N. listeri is easily de¢nable by the presence of six main signals, three of them belonging to a pyruvic acid residue and the other three originating from a symmetric substituted ribitol residue with an intensity ratio of 2:2:1 for the signals C-1,5, C-2,4 and C-3, respectively (Fig. 2g, Table 5) [60,74]. Thus, taking into account the speci¢city of 13C NMR spectra, they may be considered ¢ngerprints of species and subspecies and used fruitfully for identi¢cation of new strains of the genus Nocardiopsis. The generation of a database of the NMR spectra of other bacterial groups and development of this approach to bacterial systematics could o¡er a simple tool for identi¢cation of microorganisms in routine taxonomic practice for the future.

8. Conclusion

This review demonstrated that the structures and com- Fig. 2. 13C NMR spectra of teichoic acids found in the Nocardiopsis species and subspecies. binations of the cell wall teichoic acids, as well as products of chemical degradation of the polymers and the 13C NMR spectra, are indicative of the species and subspecies of the genus Nocardiopsis and can be used as taxonomic markers or ¢ngerprints. The data presented and the results ical signals for sugar residues in the resonance region of of studying cell wall teichoic acids in other groups of anomeric carbons (103.1 ppm for L-D-glucopyranose, Fig. Gram-positive bacteria suggest that these polymers can 2c, Table 5). be reliable taxonomic tools for support of phylogenetic delineation of bacterial taxa and for a more focused iden- 7.3. 13C NMR spectrum of teichoic acid of N. tropica ti¢cation of organisms at di¡erent taxonomic levels. Struc- tures and structural elements of cell wall teichoic acids The characteristic of the spectrum of N. tropica is the seem to be especially valuable for phenetic di¡erentiation presence of signals of ribitol phosphates and glycerol and description of species, when the traditional di¡erenti- phosphates simultaneously (Fig. 2d, Table 5), combined ating properties are variable or indistinct. Further compa- with a signal at 78.5 ppm typical of ribitol C-3 bound to rative studies of teichoic acids in the organisms of di¡erent phosphorus, as well as the signal from C-1 of glycerol-3- groups and the development of simple and e¤cient meth- phosphate (63.6^64.1 ppm) [71]. ods for the identi¢cation of the polymers, accompanied by

FEMSRE 713 1-5-01 I.B. Naumova et al. / FEMS Microbiology Reviews 25 (2001) 269^283 281 revision and approval of bacterial species and genera by [14] Fiedler, F., Seger, J., Schettenbrunner, A. and Seeliger, H.P.R. (1984) genomic and phenetic data, will be of great importance in The biochemistry of murein and cell wall teichoic acids in the genus Listeria. Syst. Appl. Microbiol. 5, 360^376. the polyphasic approach to bacterial taxonomy, which [15] Naumova, I.B. (1988) The teichoic acids of actinomycetes. Microbiol. represents the state-of-the-art approach in the unambigu- Sci. 5, 275^279. ous phenetic circumscription of taxa, and will lead to a [16] Baddiley, J. (1988) The function of teichoic acids in walls and mem- more focused identi¢cation of strains in practice. branes of bacteria. In: The Roots of Modern Biochemistry (Klein- kauf, von Dohren and Jaenicke, Eds.), pp. 223^229. Walter de Gruyter, Berlin. [17] Mauel, C., Young, M. and Karamata, D. (1991) Genes concerned Acknowledgements with synthesis of poly(glycerol phosphate), the essential teichoic acid in Bacillus subtilis strain 168, are organized in two divergent tran- This work was supported by grants from INTAS No. scription units. J. Gen. Microbiol. 137, 929^941. 96-1571 and the Russian Foundation for Basic Research [18] Mauel, C., Young, M., Monsutti-Grecescu, A., Marriott, S. and Karamata, D. (1994) Analysis of Bacillus subtilis tag gene expression No. 98-04-49277. using transcriptional fusions. Microbiology (UK) 140, 2279^2288. [19] Naumova, I.B. and Shashkov, A.S. (1997) Anionic polymers in cell walls of gram-positive bacteria. Biochemistry (Moscow) 62, 809^840. [20] Archibald, A.R. (1980) Phage receptors in gram-positive bacteria. In: References Virus Receptors (Receptors and Recognition) Series B (Randall, L.L. and Philipson, L., Eds.), Vol. 7, pp. 5^26. Chapman and Hall, Lon- [1] Goodfellow, M., Williams, S.T. and Mordarski, M. (1984) Introduc- don. tion to and importance of actinomycetes. In: The Biology of Actino- [21] Knox, K.W. and Wicken, A.J. (1973) Immunological properties of mycetes (Goodfellow, M., Mordarski, M. and Williams, S.T., Eds.), teichoic acids. Bacteriol. Rev. 37, 215^257. pp. 1^6. Academic Press, London. [22] Stackebrandt, E., Rainey, F.A. and Ward-Rainey, N.L. (1997) Pro- [2] Lechevalier, M.P. and Lechevalier, H.A. (1970) Composition of posal for a new hierarchic classi¢cation system, Actinibacteria classis whole-cell hydrolysates as a criterion in the classi¢cation of aerobic nov. Int. J. Syst. Bacteriol. 47, 479^491. actinomycetes. In: The Actinomycetales (Prauser, H., Ed.), pp. 311^ [23] Stackebrandt, E., Koch, C., Gvozdiak, O. and Schumann, P. (1995) 316. VEB Gustav Fisher Verlag, Jena. Taxonomic dissection of the genus Micrococcus: Kocuria gen. nov., [3] Goodfellow, M. (1989) Suprageneric classi¢cation of Actinomyce- Nesterenkonia gen. nov., Kytococcus gen. nov., Dermacoccus gen. tales. In: Bergey's Manual of Systematic Bacteriology (Williams, nov., and Micrococcus Cohn 1872 gen. emend. Int. J. Syst. Bacteriol. S.T., Sharpe, M.E. and Holt, J.G., Eds.), Vol. 4, pp. 2333^2339. 45, 682^692. Williams and Wilkins, Baltimore, MD. [24] Stackebrandt, E. and Prauser, H. (1991) The family Cellulomodaceae. [4] Lechevalier, M.P., DeBierve, C. and Lechevalier, H.A. (1977) Che- In: The Prokaryotes (Tru©per, H.G., Dworkin, M., Harder, W. and motaxonomy of aerobic actinomycetes: phospholipid composition. Schleifer, K.-H., Eds.), pp. 1323^1345. Springer Verlag, New Biochem. Syst. Ecol. 5, 249^260. York. [5] Collins, M.D., Pirouz, T., Goodfellow, M. and Minnikin, D.E. (1977) [25] Potekhina, N.V., Shashkov, A.S. and Naumova, I.B. (1996) The cell Distribution of menaquinones in actinomycetes and corynebacteria. wall of Actinomadura madura contains poly(galactosyl-1C2-glycerol J. Gen. Microbiol. 100, 221^230. phosphate) and poly-(3-O-methylgalactosyl-1C2-glycerol phos- [6] Kroppenstedt, R.M. (1985) Fatty acid and menaquinone analysis of phate). Microbiologiya (Moscow) 65, 522^526. actinomycetes and related organisms. Soc. Appl. Bacteriol. Tech. Ser. [26] Davison, A.L. and Baddiley, J. (1963) The distribution of teichoic 20, 173^197. acids in staphylococci. J. Gen. Microbiol. 32, 271^276. [7] Minnikin, D.E., Alshamaony, L. and Goodfellow, M. (1975) Di¡er- [27] Tul'skaya, E.M., Shashkov, A.S., Evtushenko, L.I., Bueva, O.V. and entiation of Mycobacterium, Nocardia and related taxa by thin-layer Naumova, I.B. (1997) Structural identity of teichoic acids from acti- chromatographic analysis of whole-cell methanolysates. J. Gen. Mi- nomycete species Streptomyces hygroscopicus. Biochemistry (Mos- crobiol. 88, 200^204. cow) 62, 289^293. [8] Wayne, L.G., Brenner, D.J., Colwell, R.R., Grimont, P.A.D., Kan- [28] Naumova, I.B., Kuznetsov, V.D., Kudrina, K.S. and Bezzubenkova, dler, O., Krichevsky, M.I., Moore, L.H., Moore, W.E.C., Murrey, A.P. (1980) The occurrence of teichoic acids in Streptomyces. Arch. R.G.E., Stackebrandt, E., Starr, P. and Tru«per, H.G. (1987) Report Microbiol. 126, 71^75. of the Ad Hoc Commitee on Reconciliation of Approaches to Bac- [29] Shashkov, A.S., Potekhina, N.V., Naumova, I.B., Evtushenko, L.I. terial Systematics. Int. J. Syst. Bacteriol. 37, 463^464. and Widmalm, G. (1999) Cell wall teichoic acids of Actinomadura [9] Suzuki, K., Goodfellow, M. and O'Donnel, A.G. (1994) Cell enve- viridis VKM Ac-1315T. Eur. J. Biochem. 262, 688^695. lopes and classi¢cation. In: Handbook of New Bacterial Systematics [30] Potekhina, N.V., Naumova, I.B., Shashkov, A.S. and Terekhova, (Goodfellow, M. and O'Donnel, A.G., Eds.), pp. 195^250. Academic L.P. (1991) Structural features of cell wall teichoic acid and peptido- Press, London. glycan of Actinomadura cremea INA 292. Eur. J. Biochem. 199, 313^ [10] Sutcli¡e, I.C. (1994) The lipoteichoic acids and lipoglycans of gram- 316. positive bacteria: a chemotaxonomic perspective. Syst. Appl. Micro- [31] Potekhina, N.V., Tul'skaya, E.M., Naumova, I.B., Shashkov, A.S. biol. 17, 467^480. and Evtushenko, L.I. (1993) Erythritolteichoic acid in the cell wall [11] Baddiley, J. (1972) Teichoic acids in cell walls and membranes of of Glycomyces tenuis VKM Ac-1250. Eur. J. Biochem. 218, 371^ bacteria. Essays Biochem. 8, 35^77. 375. [12] Easmon, C.S.F. and Goodfellow, M. (1990) Staphylococcus and Mi- [32] Potekhina, N.V., Tul'skaya, E.M., Shashkov, A.S., Taran, V.V., Ev- crococcus. In: Topley and Wilson's Principles of Bacteriology, Virol- tushenko, L.I. and Naumova, I.B. (1998) Species speci¢city of tei- ogy and Immunology, 8th edn. (Parker, T.M. and Duerden, B.I., choic acids in the actinomycete genus Glycomyces. Microbiologiya Eds.), pp. 161^186. Edward Arnold, London. (Moscow) 67, 330^334. [13] Fiedler, F., Scha¥er, M.J. and Stackebrandt, E. (1981) Biochemical [33] Shashkov, A.S., Tul'skaya, E.M., Evtushenko, L.I. and Naumova, and nucleic acid hybridisation studies on Brevibacterium linens and I.B. (1999) Cell wall teichoic acid of Nocardioides albus VKM Ac- related strains. Arch. Microbiol. 129, 85^93. 805. Biochemistry (Moscow) 64, 1544^1549.

FEMSRE 713 1-5-01 282 I.B. Naumova et al. / FEMS Microbiology Reviews 25 (2001) 269^283

[34] Shashkov, A.S., Tul'skaya, E.M., Evtushenko, L.I., Gratchev, A.A. actinomycete isolated from a saltern. Int. J. Syst. Evol. Microbiol. 50, and Naumova, I.B. (2000) Structure of a teichoic acid of Nocar- 1909^1913. dioides luteus VKM Ac-1246T cell wall. Biochemistry (Moscow) 65, [54] Mordarska, H., Smogov, W. and Gamian, A. (1985) Immunogenic 509^514. properties of glycolipids of Nocardiopsis dassonvillei. Arch. Immunol. [35] Meyer, J. (1976) Nocardiopsis, a new genus of the order Actinomyce- Ther. Exp. (Warsaw) 33, 523^530. tales. Int. J. Syst. Bacteriol. 26, 487^493. [55] Tsujibo, H., Sakamoto, T., Miyamoto, K., Kusano, G., Ogura, M., [36] Grund, E. and Kroppenstedt, R.M. (1990) Chemotaxonomy and nu- Hasegawa, T. and Inamori, Y. (1990) Isolation of cytotoxic sub- merical taxonomy of the genus Nocardiopsis Meyer 1976. Int. J. Syst. stance, kalafungin from an alkalophilic actinomycete, Nocardiopsis Bacteriol. 40, 5^11. dassonvillei subsp. prasina. Chem. Pharm. Bull. (Tokyo) 38, 2299^ [37] Fischer, A., Kroppenstedt, R.M. and Stackebrandt, E. (1983) Molec- 2300. ular-genetic and chemotaxonomic studies on Actinomadura and No- [56] Terekhova, L.P., Galatenko, O.A., Alferova, I.V. and Preobrazhen- cardiopsis. J. Gen. Microbiol. 129, 3433^3446. skaia, T.P. (1991) Comparative evaluation of various bacterial [38] Yassin, A.F., Galinski, E.A., Wohlfarth, A., Jahnke, K.-D., Schaal, growth inhibitors as selective agents for isolation of soil Actinomyces. K.P. and Tru«per, H.G. (1993) A new actinomycetes species, Nocar- Antibiot. Khimioter. (Moscow) 36, 5^8. diopsis lucentensis sp. nov.. Int. J. Syst. Bacteriol. 43, 266^271. [57] Kim, Y.S., Sagara, J. and Kawai, A. (1995) Studies on the antiviral [39] Yassin, A.F., Rainey, F.A., Burhardt, J., Gierth, D., Ungerechts, J., activity of protein kinase inhibitors against the replication of vesicu- Lux, I., Seifert, P., Bal, C. and Schaal, K.P. (1997) Description of lar stomatitis virus. Biol. Pharm. Bull. 18, 895^899. Nocardiopsis synnemataformans sp. nov., elevation of Nocardiopsis [58] Chiba, S., Suzuki, M. and Ando, K. (1999) Taxonomic re-evaluation alba subsp. prasina to Nocardiopsis prasina comb. nov., and designa- of Nocardiopsis sp. K-252T ( = NRRL 15532T): a proposal to trans- tion of Nocardiopsis antarctica and Nocardiopsis alborubida as later fer this strain to the genus Nonomuraea as Nonomuraea longicatena subjective synonyms of Nocardiopsis dassonvillei. Int. J. Syst. Bacter- sp. nov. J. Syst. Bacteriol. 49, 1623^1630. iol. 47, 983^988. [59] Newton, G.L., Arnold, K., Price, M.S., Sherrill, C., Delcardayre, [40] Evtushenko, L.I., Taran, V.V., Akimov, V.N., Kroppenstedt, R., S.B., Aharonowitz, Y., Cohen, G., Davies, J., Fahey, R.C. and Da- Tiedje, J.M. and Stackebrandt, E. (2000) Nocardiopsis tropica sp. vis, C. (1996) Distribution of thiols in microorganisms: mycothiol is a nov., Nocardiopsis trehalosei sp. nov., nom. rev., and subspecies No- major thiol in most actinomycetes. J. Bacteriol. 178, 1990^1995. cardiopsis dassonvillei subsp. albirubida subsp. nov., comb. nov.. Int. [60] Streshinskaya, G.M., Tul'skaya, E.M., Shashkov, A.S., Evtushenko, J. Syst. Bacteriol. 50, 73^81. L.I., Taran, V.V. and Naumova, I.B. (1998) Teichoic acids of the cell [41] Rainey, F.A., Ward-Rainey, N., Kroppenstedt, R.M. and Stacke- wall of Nocardiopsis listeri, Nocardiopsis lucentensis, and Nocardiop- brandt, E. (1996) The genus Nocardiopsis represents a phylogeneti- sis tregalosei. Biochemistry (Moscow) 63, 230^234. cally coherent taxon and a distinct actinomycete lineage: proposal of [61] Archibald, A.R. (1974) The structure, biosynthesis and function of Nocardiopsaceae fam. nov. Int. J. Syst. Bacteriol. 46, 1088^1092. teichoic acid. Adv. Microbiol. Physiol. 11, 53^95. [42] Al-Tai, A.M. and Ruan, J.-S. (1994) Nocardiopsis halophila sp. nov., [62] Tul'skaya, E.M., Vylegzhanina, K.S., Streshinskaya, G.M., Shash- a new halophilic actinomycete isolated from soil. Int. J. Syst. Bacter- kov, A.S. and Naumova, I.B. (1991) 1,3-Poly(glycerol phosphate) iol. 44, 474^478. chains in the cell wall of Streptomyces rutgersensis subsp. castelarense [43] Gordon, R.E. and Horan, A.S. (1968) Nocardia dassonvillei, a macro- VKM Ac-238. Biochim. Biophys. Acta 1074, 237^242. scopic replica of Streptomyces griseus. J. Gen. Microbiol. 50, 235^ [63] Streshinskaya, G.M., Naumova, I.B., Shashkov, A.S., Kozlova, 240. Yu.I., Terekhova, L.P. and Galatenko, O.A. (1991) Some structural [44] Miyashita, M., Mikami, Y. and Arai, T. (1984) Alkalophilic actino- peculiarities of cell wall polymers of Actinomadura (Nonomuraea) mycete, Nocardiopsis dassonvillei subsp. prasina subsp. nov. isolated polychroma INA 2755. Biochemistry (Moscow) 56, 2270^2280. from soil. Int. J. Syst. Bacteriol. 34, 405^409. [64] Kozlova, Yu.I., Streshinskaya, G.M., Shashkov, A.S., Evtushenko, [45] Kroppenstedt, R.M. (1992) The genus Nocardiopsis In: The Prokary- L.I. and Naumova, I.B. (1999) Anionic carbohydrate-containing pol- otes, 2nd edn. (Balows, A., Tru«per, H.G., Dworkin, M., Harder, W. ymers of cell walls in two streptoverticille genospecies. Biochemistry and Schleifer, K.-H., Eds.), pp. 1139^1156. Springer Verlag, New (Moscow) 64, 671^677. York. [65] Sadikov, B.M., Naumova, I.B., Streshinskaya, G.M. and Polin, A.N. [46] Brocg-Rousseau, D. (1904) Sur un Streptothrix. Ref. Gen. Bot. 16, (1987) Cell wall polymers containing carbohydrates of Arthrobacter 219^230. species. Microbiology (Moscow) 56, 441^446. [47] Beau, F., Bollet, C., Coton, T., Garnotel, E. and Drancourt, M. [66] Shashkov, A.S., Tul'skaya, E.M., Grachev, A.A., Evtushenko, L.I., (1999) Molecular identi¢cation of a Nocardiopsis dassonvillei blood Bueva, O.A. and Naumova, I.B. (1998) Structure of teichoic acid of isolate. J. Clin. Microbiol. 37, 3366^3368. the cell wall of Streptomyces sparsogenes VKM Ac-1744T. Biochem- [48] Sindhuphak, W., Macdonald, E. and Head, E. (1985) Actinomyceto- istry (Moscow) 63, 1098^1103. ma caused by Nocardiopsis dassonvillei. Arch. Dermatol. 121, 1332^ [67] Fiedler, F. and Bude, A. (1989) Occurrence and chemistry of cell wall 1334. teichoic acids in the genus Brevibacterium. J. Gen. Microbiol. 135, [49] Mordarska, H., Zakrzewska-Czerwinska, J., Pasciak, M., Szponar, B. 2837^2846. and Rowinski, S. (1998) Rare, suppurative pulmonary infection [68] De Boer, W.R., Wouters, J.T.M., Anderson, A.J. and Archibald, caused by Nocardiopsis dassonvillei recognized by glycolipid markers. A.R. (1978) Further evidence for the structure of the teichoic acids FEMS Immunol. Med. Microbiol. 21, 47^55. from Bacillus stearothermophilus B65 and Bacillus subtilis var. niger [50] Dolak, L.A., Castle, T.M. and Laborde, L.A. (1981) Biologically WM. Eur. J. Biochem. 85, 433^436. pure culture of Nocardiopsis trehalosei sp. nov. US Patent 4,306,028. [69] Shashkov, A.S., Zaretskaya, M.Sh., Yarotsky, S.V., Naumova, I.B., [51] Abyzov, S.S., Philipova, S.N. and Kuznetsov, V.D. (1983) Nocardiop- Chizhov, O.S. and Shabarova, Z.A. (1979) On the structure of the sis antarcticus, a new species of actinomycetes, isolated from the ice teichoic acid from the cell wall of Streptomyces antibioticus 39. Local- sheet of the central antarctic glacier. Izv. Akad. Nauk SSSR Ser. ization of the phosphodiester linkages and elucidation of the mono- Biol. 4, 559^568. meric units structure by means of 13C NMR spectroscopy. Eur. J. [52] Andersson, A.M., Weiss, N., Rainey, F. and Salkinoja-Salonen, M.S. Biochem. 102, 477^481. (1999) Dust-borne bacteria in animal sheds, schools and children's [70] Uchikawa, K., Sekikawa, I. and Azuma, I. (1986) Structural studies day care centres. J. Appl. Microbiol. 86, 622^634. on teichoic acids in cell walls of several serotypes of Listeria mono- [53] Chun, J., Bae, K.S., Moon, E.Y., Jung, S.O., Lee, H.K. and Kim, cytogenes. J. Biochemistry 99, 315^327. S.J. (2000) Nocardiopsis kunsanensis sp. nov., a moderately halophilic [71] Streshinskaya, G.M., Kozlova, Yu.I., Evtushenko, L.I., Taran, V.V.,

FEMSRE 713 1-5-01 I.B. Naumova et al. / FEMS Microbiology Reviews 25 (2001) 269^283 283

Shashkov, A.S. and Naumova, I.B. (1996) Cell wall teichoic acid of phosphate) teichoic acid in the wall of Bacillus stearothermophilus Nocardiopsis subsp. VKM Ac-1457. Biochemistry (Moscow) 61, 285^ B65. Biochem. J. 151, 115^120. 288. [83] Kojima, N., Uchikawa, K., Araki, Y. and Ito, E. (1986) Structural [72] Streshinskaya, G.M., Shashkov, A.S. and Naumova, I.B. (1995) Het- studies on the minor teichoic acid of Bacillus coagulans AHU 1631. erogeneity of the chains of teichoic acids from the cell wall of Strep- Eur. J. Biochem. 155, 521^526. tomyces chrysomallus VKM Ac-628. Biochemistry (Moscow) 60, 963^ [84] Kozlova, Yu.I., Streshinskaya, G.M., Shashkov, A.S., Evtushenko, 969. L.I., Gavrish, E.Yu. and Naumova, I.B. (2000) Structure of carbohy- [73] Potekhina, N.V., Naumova, I.B., Shashkov, A.S. and Kuznetsov, drate-containing cell wall polymers in several representatives of the V.D. (1992) 3-O-Methylrhamnose within the composition of the tei- order Actinomycetales. Biochemistry (Moscow) 65, 1432^1439. choic acid of the Streptomyces roseolus ISP 5174 cell wall. Biochem- [85] Kozlova, Yu.I., Streshinskaya, G.M., Shashkov, A.S., Evtushenko, istry (Moscow) 57, 1206^1214. L.I. and Naumova, I.B. (1996) Poly(glucosylglycerol phosphate) in [74] Tul'skaya, E.M., Shashkov, A.S., Evtushenko, L.I., Taran, V.V. and cell walls of Streptomyces £avotricini VKM Ac-1277. Biochemistry Naumova, I.B. (1995) Novel cell-wall teichoic acid from Nocardiopsis (Moscow) 61, 1892^1897. albus subsp. albus as a species-speci¢c marker. Microbiology (UK) [86] Schipper, D. (1995) Structural studies of the teichoic acids from Ba- 141, 1851^1856. cillus licheniformis. Carbohydr. Res. 279, 75^82. [75] Tul'skaya, E.M., Shashkov, A.S., Evtushenko, L.I. and Naumova, [87] Shashkov, A.S., Malysheva, V.A., Naumova, I.B., Streshinskaya, I.B. (2000) Cell wall teichoic acids of Nocardiopsis prasina VKM G.M. and Evtushenko, L.I. (1993) Poly(ribofuranosylribitol phos- Ac-1880T. Microbiology (Moscow) 69, 48^50. phate) in cell wall of Agromyces cerinus subsp. nitratus VKM Ac- [76] Schubert, K., Reiml, D., Accolas, J.-P. and Fiedler, F. (1993) A novel 1351. Bioorg. Khim. (Moscow) 19, 433^438. type of meso-diaminopimelic acid-based peptidoglycan and novel [88] Fischer, W., Behr, T., Hartmann, R., Peter-Katalinic, J. and Egge, H. poly(erythritol phosphate) teichoic acids in cell walls of two coryne- (1993) Teichoic acid and lipoteichoic acid of Streptococcus pneumo- form isolates from the surface £ora of French cooked cheeses. Arch. niae possess identical chain structures. A reinvestigation of teichoic Microbiol. 160, 222^228. acid (C polysaccharide). Eur. J. Biochem. 215, 851^857. [77] Schubert, K., Ludwig, W., Springer, N., Kroppenstedt, R.M., Acco- [89] Abeygunawardana, Ch., Bush, C.A. and Cisar, J.O. (1991) Complete las, J.P. and Fiedler, F. (1996) Two coryneform bacteria isolated structure of the cell surface polysaccharide of Streptococcus oralis from the surface of French Gruyere and Beaufort cheeses are new C104: A 600-MHz NMR study. Biochemistry 30, 8568^8577. species of the genus Brachybacterium: Brachybacterium alimentarium [90] Endle, J., Seidle, P.H., Fiedler, F. and Schleifer, K.H. (1983) Chem- sp. nov. and Brachybacterium tyrofermentans sp. nov. Int. J. Syst. ical composition and structure of cell wall teichoic acids of staph- Bacteriol. 46, 81^87. ylococci. Arch. Microbiol. 135, 215^223. [78] Anderton, W.J. and Wilkinson, S.G. (1985) Structural studies of a [91] Tul'skaya, E.M., Streshinskaya, G.M., Naumova, I.B., Shashkov, mannitol teichoic acid from cell wall of bacterium NCTC 9742. Bio- A.S. and Terekhova, L.P. (1993) A new structural type of teichoic chem. J. 226, 587^599. acid and some chemotaxonomic criteria of two species Nocardiopsis [79] Veerkamp, J.H., Hoelen, G.J.M. and Op Den Camp, H.J.M. (1983) dassonvillei and Nocardiopsis antarcticus. Arch. Microbiol. 160, 299^ The structure of a mannitol teichoic acid from Bi¢dobacterium bi¢- 305. dum ssp. pennsylvanicum. Biochim. Biophys. Acta 755, 439^451. [92] Shashkov, A.S., Streshinskaya, G.M., Kozlova, Yu.I., Potekhina, [80] Shashkov, A.S., Streshinskaya, G.M., Gnilozub, V.A., Evtushenko, N.V., Evtushenko, L.I., Taran, V.V. and Naumova, I.B. (1997) Struc- L.I. and Naumova, I.B. (1995) Poly(arabitol phosphate) teichoic acid ture of teichoic acid from cell walls of Nocardiopsis alborubida. Bio- in the cell wall of Agromyces cerinus subsp. cerinus VKM Ac-1340T. chemistry (Moscow) 62, 1135^1139. FEBS Lett. 371, 163^166. [93] Streshinskaya, G.M., Shashkov, A.S., Evtushenko, L.I. and Naumo- [81] Fischer, W. (1988) Physiology of lipoteichoic acids in bacteria. Adv. va, I.B. (2000) Cell wall anionic polymers in rare genera of the order Microb. Physiol. 29, 233^302. Actinomycetales. In: Int. Symp. `Modern Problems of Microbial Bio- [82] Anderson, A.J. and Archibald, A.R. (1975) Poly(glucosylglycerol chemistry and Biotechnology', Pushchino, p. 82.

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