Biological and chemical comparison of butanol‐ and phenol‐water extracted lipopolysaccharide from Capnocytophaga sputigena

Lipopolysacharide was isolated form Capnocytophaga sputigena strain 4 by the butanol‐water method, and its release and association with acid (AcP) and alkaline (AIP) phosphatase was compared. Approximately 70% of the AcP and AIP was released with the butanol‐water extracted LPS (BLPS), with one‐third of the released phosp0hatases co‐eluting with it on Sepharose 4B. The BLPS was low in carbohydrate, KDO, lipid, heptose, hexosamine, and higher in phosphate and protein that the phenol‐water (PLPS) extracted LPS, SDS‐PAGE‐periodic acid Schiff staining revealed carbohydreate in both LPS preparations, while Commassie Blue staining revealed peptides only in BLPS, occurring as several minor bands and one major band at 39–41 kilodaltons. Immunoferritin reactions localized the LPS at the surface of intact C. sputigena; both LPSs were, however, immunochemically distinct. Incorporation of 3H‐thymidine into BLPS stimulated whole and T‐depleted C3H/HeJ mouse spleenocytes 7 and 4 fold, respectively higher than with PLPS. A phosphatase‐LPS interaction employing purified molecules was examined.