成团泛菌低分子脂多糖的分离纯化和鉴定

目的 :从革兰阴性非致病菌成团泛菌中提取、分离和纯化出细菌脂多糖 ,开发一种新的免疫调节剂 Pantoea agglomeranslipopolysaccharide(L PSp)。方法 :用 westphal热酚水法粗提 ,阴离子交换层析法进行纯化。应用 Tricine- SDS-聚丙稀酰胺凝胶电泳法 (Tricine- SDS- PAGE)、鲎变形细胞溶解物实验法对 L PSp的相对分子质量及活性进行鉴定。同时对 L PSp的化学成分进行分析。结果 :获得相对分子质量为 5 0 0 0的低分子脂多糖 (L PSp)。经鉴定 L PSp为含有 1 8%2 - keto- 3- deoxyoctulosonate (KDO)、总糖量为 6 4 .5 %、总纯度 >99.0 7%、重量比活性较 E.coli O1 1 1 :B4 L PS高 2 .6倍。结论 :用阴离子交换层析法可纯化出低相对分子质量高纯度水溶性的成团泛菌脂多糖 L PSp,与其它细菌 L PS相比 L PSp具有较高的 KDO含量

PURIFICATION AND IDENTIFICATION OF LPS PREPARED FROM PANTOEA AGGLOMERANS

Objective:To explore a novel immune regulator (LPSp) derived from lipopolysaccharide of one avirulent gram negative bacteria Pantoea agglo merans. Methods:The primary LPSp was extracted with hot phenol method, then purified by anion exchange chromatography. The molecular weight and bioactivity of LPSp were identified by Tricine sodium dodecyl sulfate polyacrylamide gel electrophoresis(Tricine SDS PAGE) and limulus amebocyte lysate test respectively. The chemical components of LPSp were also analyzed in our study.Result:The purified LPSp being characterized with a molecular of 5 000,18% of 2 keto 3 deoxyoctulosonate (KDO), and 64% of total sugar, purified >99.07%, 2 6 times of relative biological activities higher than those of LPS from E.coli O111:B4.Conclusion:Solvable LPSp with low molecular weight and high value of KDO can be purified from Pantoea agglomerans efficiently by Q Sepharose FF anion exchange chromatography. The KDO content of LPSp was higher than some other bacteria.