QUANTITATIVE EXTRACTION AND PURIFICATION OF EXOPOLYSACCHARIDES FROM KLEBSIELLA-PNEUMONIAE
QUANTITATIVE EXTRACTION AND PURIFICATION OF EXOPOLYSACCHARIDES FROM KLEBSIELLA-PNEUMONIAE
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DOI：
10.1016/0167-7012(89)90038-9
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发表时间：
1989-03-01
影响因子：
2.2
通讯作者：
CUNHA, BA
中科院分区：
文献类型：
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作者：
DOMENICO, P;DIEDRICH, DL;CUNHA, BA
在柠檬酸盐缓冲液（pH 4.5）中用两性离子去污剂提取肺炎克雷伯菌（O 1：K2）胞外多糖。温和的程序释放了近95%的荚膜多糖和60-70%的脂多糖。用多糖提取了不同量的蛋白质，而在提取物中发现了非常少的核酸。多糖提取物的凝胶过滤色谱法，没有缓冲液交换步骤，导致高回收率纯化的荚膜多糖和脂多糖。提取导致最小的细胞破坏，如通过光学和电子显微镜、通过活力研究和通过质粒DNA分离所示。由于与多糖的相互作用，在去污剂提取之前分离的质粒在琼脂糖凝胶上被看到涂抹，而来自提取的细胞的质粒表现出紧密的带型。离子相互作用和亲水相互作用似乎是K的结构稳定性的原因。肺炎荚膜
Klebsiella pneumoniae (O1:K2) exopolysaccharides were extracted with a zwitterionic detergent in citrate buffer (pH 4.5). The mild procedure released nearly 95% of the capsular polysaccharide and 60-70% of the lipopolysaccharide. Variable amounts of protein were extracted with the polysaccharides, while very little of the nucleic acid was found in the extract. Gel filtration chromatography of the polysaccharide extract, without a buffer exchange step, resulted in high recovery purification of capsular and lipopolysaccharides. The extraction led to minimal cell disruption, as shown by light and electron microscopy, by studies of viability, and by plasmid DNA isolation. Plasmids isolated before detergent extraction were seen to smear on agarose gels, due to interaction with polysaccharides, while plasmids from extracted cells exhibited tight banding patterns. Both ionic and hydrophic interactions appear responsible for the structural stability of the K. pneumoniae capsule.