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肺炎克雷伯氏菌荚膜多糖的提取纯化及其对细胞免疫活性的影响
引用本文:张燕, 李轻舟, 杜连祥, 戚薇, 陈锦英,.肺炎克雷伯氏菌荚膜多糖的提取纯化及其对细胞免疫活性的影响[J].生物工程学报,2005,21(3):461-465.
作者姓名:张燕  李轻舟  杜连祥  戚薇  陈锦英  
作者单位:1. 天津科技大学生物工程学院,天津,300222
2. 天津医科大学医学微生物学院,天津,300070
摘    要:采用液体深层培养得肺炎克雷伯氏菌Kp9株发酵液,研究确立了菌体快速裂解条件:NP40 1%和胰蛋白酶25 0IUg菌体,50℃作用1h ,然后加入溶菌酶80μg/mL ,56℃作用1h ;裂解液经超滤浓缩和有机溶剂沉淀获得多糖粗品;多糖粗品先后经CTAB吸附分离,DEAE-SepharoseFastFlow离子交换和SephacrylS 30 0HR凝胶过滤纯化,得分子量分布相对均一的多糖纯品,产品得率为0.25 1g/L。采用淋巴细胞转化实验分别探讨了多糖粗品和纯品的免疫活性,研究显示荚膜多糖具有高效的体外细胞免疫活性,并具有典型的双向免疫调节作用。研究结论为肺炎克雷伯氏菌荚膜多糖的开发奠定了前期基础。

关 键 词:肺炎克雷伯氏菌    荚膜多糖    提取    纯化    细胞免疫活性  
文章编号:1000-3061(2005)03-0461-05
修稿时间:2004年11月1日

Extraction and Purification of the Klebsiella pneumoniae Capsular Polysaccharide and the Effection on the Cell Immunoactivity
ZHANG Yan,LI Qing-Zhou,DU Lian-xiang,QI Wei,CHEN Jin-ying.Extraction and Purification of the Klebsiella pneumoniae Capsular Polysaccharide and the Effection on the Cell Immunoactivity[J].Chinese Journal of Biotechnology,2005,21(3):461-465.
Authors:ZHANG Yan  LI Qing-Zhou  DU Lian-xiang  QI Wei  CHEN Jin-ying
Institution:College of Bioengineering, Tianjin University of Science and Technology, Tianjin 300222, China. yzhang@tust.edu.cn
Abstract:Klebsiella pneumoniae was cultured followed by the preparation and immunoactivity elucidating of its polysaccharide(CPS).The lysis of cell is the first key step in the preparation,under the co-action of trypsin, lysozyme and NP-40,the cell lysed within 2h,then the lysate was concentrated by ultrafiltration which serves as concentrating and partial purifying action simultaneously.Crude CPS was got by ethanol precipitation,then purified through the Ion-exchange and gel filtration,the purity of CPS was judged by the gel filtration and agarose gel electrophoresis.The effect of CPS on the cell immunoactivity was studied in detail,the results show that CPS possesses bidirectional immunoregulation on the spleen cells of mice,that is,low concentration of CPS can stimulate the immune response while the high concentration manifests the inhibition significantly.The investigation results will benefit on the exploitation of the CPS.
Keywords:Klebsiella pneumoniae  polysaccharide  extraction  purification  immunoactivity
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