Teichoic acid from the walls of Staphylococcus aureus H. 2. Location of phosphate and alanine residues

Abstract

The structure of the ribitol teichoic acid from the walls of Staphylococcus aureus H was established by periodate oxidation and alkali hydrolysis. Periodate oxidation after removal of alanine together with determination of formaldehyde produced indicates that the 4-O-(N-acetylglucosaminyl)-D-ribitol units are joined through 1,5-phosphodiester linkages. Titra-tion confirms that eight units are present. Periodate oxidation before and after removal of D-alanine ester residues establishes that these are attached through hydroxyl groups at positions 2 or 3 on ribitol. Alkali hydrolysis gave a mixture of 4-O-glucosaminylribitol 1- and 2-phosphates possessing both [alpha]- and [beta]-glycosidic linkages. Ion-exchange chromatography caused separation into [alpha] and [beta] isomers, whereas paper chromatography separated the 2- and 1-phosphate isomers. The isomeric phosphates were degraded to chitose and the respective ribitol phosphates (without phosphate migration) by treatment with nitrous acid. The proportion of [alpha]- to [beta]-glycosidic linkages varies in preparations from different batches of organism. A compound formed during the prolonged action of hot hydrochloric acid on ribitol derivatives (previously called "dianhydroribitol") is believed to be a 5-chloro derivative of anhydroribitol.