2All-Russian Collection of Microorganisms, Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 142292 Pushchino, Moscow Region, Russia; fax: (495) 923-3602
3Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky pr. 47, 119992 Moscow, Russia; fax: (499) 135-5328
4Institute of Ecology and Genetics of Microorganisms, Ural Branch of the Russian Academy of Sciences, ul. Goleva 13, 614081 Perm’, Russia; fax: (342) 244-6711
* To whom correspondence should be addressed.
Received July 11, 2008; Revision received October 20, 2008
Cell walls of Bacillus subtilis VKM B-760 and VKM B-764 are characterized by heterogeneous composition of teichoic acids. Polymer I with structure -6)-β-D-Galp-(1→1)-sn-Gro-(3-P-, polymer II with structure -6)-α-D-Glcp-(1→1)-sn-Gro-(3-P-, and a small amount of unsubstituted 1,3-poly(glycerol phosphate) were detected in strain VKM B-760. Strain VKM B-764 contains an analogous set of teichoic acids, but a characteristic feature of polymer II is the presence of disubstituted glycerol residue with α-glucopyranose localization in the integral chain at C-1 hydroxyl and β-glucopyranose as a side branch at C-2 hydroxyl (polymer III): -6)-α-D-Glcp-(1→1)-[β-D-Glcp-(1→2)]-sn-Gro-(3-P-. The structures of polymer I in bacilli and polymer III in Gram-positive bacteria are described for the first time. Teichoic acids were studied by chemical methods and on the basis of combined analysis of one-dimensional 1H-, 13C-, and 31P-NMR spectra, homonuclear two-dimensional 1H/1H COSY, TOCSY, and ROESY, and heteronuclear two-dimensional 1H/13C gHSQC- and HMQC-TOCSY experiments. Simultaneous presence of several different structure teichoic acids in the bacillus cell walls as well as chemotaxonomical perspectives of the application of these polymers as species-specific markers for members of the Bacillus genus is discussed.
KEY WORDS: Bacillus subtilis, cell wall, teichoic acids, NMR spectroscopy