Analysis of lipopolysaccharides of Pasteurella multocida and several Gram-negative bacteria by gas chromatography on a capillary column

Ryu, Hyo-ik; Kim, Chul-joong

Korean Journal of Veterinary Research 2000;40(1):72-80.

Hyo-ik Ryu¹, Chul-joong Kim²

¹Department of Enteric Infections, Walter Reed Army Institute of Research
²College of Veterinary Medicine, Chungnam National University

Gas chromatography의 capillary column을 이용한 Pasteurella multocida 및 기타 그람음성 세균의 lipopolysaccharide 분석

류효익¹, 김철중²

¹Walter Reed 미군 연구소, 장내감염부
²충남대학교 수의과대학

Abstract

Lipopolysaccharides (LPS) of Pasteurella multocida (P multocida) and several Gram-negative bacterial pathogens were analyzed by methanolysis, trifluoroacetylation and gas chromatography (GC) on a fused-silica capillary column. The GC analysis indicated that LPS prepared from a strain of P multocida by phenol-water (PW) or trichloroacetic acid (TCA) extraction were quite different in chemical composition. However, LPS prepared from Salmonella enteritidis by the two extraction methods were very similar. PW-LPS extracts from different Pasteurella strains of a serotype had essentially identical GC patterns. Endotoxic LPS extracted from 16 different serotypes of P multocida by PW or by phenol-chloroform-petroleum ether procedures yielded chromatograms indicating similar composition of the fatty acid moieties but minor differences in carbohydrate content. When the chemical composition of endotoxic LPS extracted from several Gram-negative bacteria (P multocida, Pasteurella hacmolytica, Haemophilus somnus, Actinobacillus ligniersii, Brucella abortus, Treponema hyodysenteriae, Escherichia coli, Bacteriodes fragilis, Salmonella abortus equi and Salmonella enteritidis) were examined, each bacteria showed a unique GC pattern. The carbohydrate constituents in LPS of various Gram-negative bacteria were quite variable not only in the O-specific polysaccharides but also in the core polysaccharides. The LPS of closely related bacteria shared more fatty acid constituents with each other than with unrelated bacteria.

Key Words: lipopolysaccharides, Pasteurella multocida, gas chromatography