Arch Microbiol (1989) 152:317 -321 Archives ol tlisroIiolory @ Springer-Verlag 1989 Madurose is often present in small amounts only, and frequently also difficult to identifity. We have therefore re- cently developed a HPLC method which permits the analysis of strains with low madurose levels (Yokota and Hasegawa 1988, 1989). Using this method, we have been searching for madurose-containing strains in the genus Streptomyces, and until now we have found five strains in two species of Streptomyces which contain madurose as a whole-cell sugar. The present paper describes the distribution of strains with madurose in the gertus Streptomyces, and also the identification of madurose from Streptomyces platensis. Materials and methods Bacterial strains and culture conditions A11 the strains of actinomycetes used in this study are from the culture collection of this institute (IFO). The type strain is shown with the superscript "T". Strains were cultured in a medium containing 1.o/o yeast extract arrd 1o/o o-glucose (pH 7.0) at 28" C for 4 days with shaking. The cells were washed with distilled water and lyophilized. Detection of madurose in the whole-cell hydrolysate The dried cells (100 mg) were hydrolyzed with 4 M HCI at 100'C for 2h in a screw-capped test tube. After filtration, the hydrolysate was concentrated in vacuo. The residue was dissolved in distilled water and neutralized with 5 M NaOH; water was added to make 1 ml. The madurose content in the hydrolysate was determined by HPLC as described in previous papers (Yokota and Hasegawa 1988, 1989). Isolation of madurose The dried cells (3 g) of S. platensis IFO 14008 were hydrolyzed with 4 M HCI at 100'C for 2 h. The hydrolysate was filtered and concentrated in vacuo, then passed through columns of IR 120 (H+) and IRA 400 (OH-), and finally concentrated in vacuo. The residue was chromatographed on a column of cellulose powder (1.8 x 75 cm) with solvent (i) (see Analytical methods). Fractions of 5 ml each were collected; the madurose-containing fractions (No. 30-45) were combined, concentrated in vacuo, and the residue was further purified by chromatography on paper with solvent (i). The madurose-containing band on the paper was ex- Streptomyces species with madurose (3-O-methyl-D-galactose) as a whole-cell sugar A. Yokota, Y. Nakagaito, and T. Hasegawa Institute for Fermentation, Osaka, 17 -85, Juso-honmachi 2-chome, Yodogawa-ku, Osaka 532, Japan Abstract. Madurose, an actinomycete whole-cell sugag was found in the strains of the genus Streptomyces: three strains of S. p latensis, one strain each of S. platezsr,s subsp. malvinus, and S. albus subsp. albus. The sugar was isolated from the hydrolysate of S. platensis IFO 14008 cells, and was ident- ified as madurose (3-O-methyl-o-galactose) by chromatog- raphic analyses, 1H-NMR spectrometry, mass spectrometry as its alditol acetate, and demethylation with boron tri- chloride. Cell walls of the strain contained peptidoglycan and teichoic acids. rr-Diaminopimelic acid, glycine, glu- tamic acid, and alanine were present in the peptidoglycan fraction in molar ratios of 1.0:1.3:7.2:2.3. Madurose was detected in the teichoic acid fraction, which was composed of phosphorus, glycerol, galactose, and madurose in molar ratios of 9.3:8.5:2.9:1.0. Thus, madurose was found in the glycerol teichoic acid moiety of the cell walls of this strain. Key words: Streptomyces - Streptomyces platensis - Madurose - 3-O-Methyl->galactose - Teichoic acid - Chemotaxonomy - Peptidoglycan Cell-wall type and whole-cell sugar pattern have become widely used as chemotaxonomic markers to differentiate distinct genera of actinomycete (Lechevalier and Lechevalier 1965). Until now, twelve genera of aerobic actinomycetes are known to possess the madurose-containing whole-cell sugar pattern (cell-wall type III/B); they include Actinomadur a, Dermatop hilus, Excellospora, " N onomuria", Micr obisp ora, Micr ote tr aspor a, Planobispora, P lanomono- spora, Spirillospora, " Streptomycoides", Streptosporangium and Thermomonospora (Lechevalier and Lechevalier 1980; Hasegawa et al. 1986; Itoh et al. 1987; Goodfellow et al. 1988). Furthermore, madurose-containing actinomycete strains with cell-wall type II/D (Miuomonospora carbonacea stbsp. carbonacea, "M. chalcea sobsp. izumensis" and M. rosaria) (Meyertons et al. 1988) and with cell-wall type IV/ A (Kibdelosporangium aridum) (Shearer et al. 1986) have recently been reported. Offprint requests ro.' A. Yokota Abbreviations: PC, paper chromatography; TLC, thin-layer chro- matography; HPLC, high performance liquid chromatography; GLC, gas-liquid chromatography ; TCA, trichloroacetic acid