Kondo S, Hisatsune K
Department of Microbiology, School of Pharmaceutical Sciences, Josai University, Saitama.
Microbiol Immunol. 1988;32(9):907-15. doi: 10.1111/j.1348-0421.1988.tb01452.x.
A simple and rapid method was devised for direct isolation and fractionation of the "degraded polysaccharide" (DPS) fraction of O-antigenic (or endotoxic) lipopolysaccharides (LPS) directly from heat-killed Vibrio cholerae (O1 and non-O1) cells without separating the LPS. Neither phenol-water extraction nor ultracentrifuge is needed in this method. V. cholerae NIH 41 was used as standard. The cells (3-5 g wet weight) were heated in 5% acetic acid at 100 C for 1.5 hr. The acetic acid extract obtained as the supernatant by centrifugation was evaporated to dryness in vacuo, and the resultant residue was dissolved in 10 ml of distilled water. The solution was mixed with 2 volumes of acetone, and the supernatant obtained by centrifugation was mixed with 5 volumes of acetone and centrifuged. Fraction Sed. II was recovered as the precipitate, while the supernatant was evaporated to dryness in vacuo, yielding fraction Sup. III. Sed. II had a sugar composition that was identical, at least qualitatively, to that of DPS isolated from LPS of the corresponding strain except for the absence of a fructose component in the case of V. cholerae NIH 41, while instead Sup. III from V. cholerae NIH 41 contained fructose.
设计了一种简单快速的方法，可直接从热灭活的霍乱弧菌（O1和非O1）细胞中直接分离和分级O抗原（或内毒素）脂多糖（LPS）的“降解多糖”（DPS）部分，而无需分离LPS。该方法既不需要酚水提取，也不需要超速离心。以霍乱弧菌NIH 41作为标准菌株。将细胞（3-5克湿重）在5%乙酸中于100℃加热1.5小时。通过离心获得的作为上清液的乙酸提取物在真空中蒸发至干，所得残渣溶解于10毫升蒸馏水中。将该溶液与2倍体积的丙酮混合，离心得到的上清液再与5倍体积的丙酮混合并离心。沉淀部分回收得到沉降物II，而上清液在真空中蒸发至干，得到上清液III部分。沉降物II的糖组成至少在定性上与从相应菌株的LPS中分离出的DPS相同，只是霍乱弧菌NIH 41的情况除外，其缺少果糖成分，而霍乱弧菌NIH 41的上清液III部分含有果糖。
