长双歧杆菌BBMN68胞外多糖生物合成基因的克隆及生物信息学分析

发布时间：2018-08-23 19:41

【摘要】：目的:研究长双歧杆菌BBMN68胞外多糖生物合成途径和调控机制。方法:提取BBMN68胞外多糖,预测eps基因簇各基因编码蛋白功能,克隆关键基因并对其进行进化分析。结果:在CDM和MRSC培养基上,长双歧杆菌BBMN68产生胞外多糖,产量分别为100 mg/L和400 mg/L。本实验室工作人员对其进行全基因组序列测定。根据全基因组序列,分析得到eps基因簇,对该基因簇各基因编码蛋白的功能预测发现,BBMN68_1002编码1个可能具有多糖重复单元转运子功能的蛋白。BBMN68_1003、BBMN68_1006、BBMN68_1007和BMN68_1011与多糖重复单元聚合有关。BBMN68_1004、BBMN68_1008和BBMN68_1009参与糖基转移。BMN68_1010控制多糖分子链长。BBMN68_1012编码胞外多糖合成的关键酶——引导糖基转移酶(priming glycosyltransferase,Cps D),启动合成胞外多糖第1步。由Cps D基因构建的系统发育树,BBMN68与猿猴、狨猴及大猩猩的长双歧杆菌同枝,可能是饮食影响肠道细菌进化的结果。结论:本研究结果为解析长双歧杆菌BBMN68胞外多糖生物合成途径和调控机制奠定了理论基础。
[Abstract]:Objective: to study the biosynthesis pathway and regulation mechanism of BBMN68 exopolysaccharide from Bifidobacterium longii. Methods: extracellular polysaccharides of BBMN68 were extracted, protein function of eps gene cluster was predicted, and key genes were cloned and analyzed. Results: on CDM and MRSC medium, BBMN68 produced extracellular polysaccharides with the yield of 100 mg/L and 400 mg/L., respectively. The whole genome sequence was sequenced by our laboratory staff. According to the whole genome sequence, the eps gene cluster was obtained. It is found that BBMN681002 encodes a protein. BBMN681003 / BBMN681003 / BBMN681006 BBMN686BBMN681007 and BMN68_1011 are associated with polysaccharide repeat unit polymerization. BBMN684BBMN681008 and BBMN68_1009 participate in glycosyl transfer. BMN681010 control polysaccharides. Molecular chain length. BBMN681012 encodes the key enzyme of extracellular polysaccharide synthesis-leading glycosyltransferase (priming glycosyltransferase,Cps D),) to initiate the synthesis of extracellular polysaccharides. BBMN68, a phylogenetic tree constructed from Cps D gene, shares the same branch with bifidobacteria in apes, marmosets and gorillas, which may be the result of diet affecting the evolution of intestinal bacteria. Conclusion: this study provides a theoretical basis for elucidating the biosynthesis pathway and regulation mechanism of BBMN68 exopolysaccharide in Bifidobacterium longiferi.
【作者单位】：中国农业大学食品科学与营养工程学院食品质量与安全北京实验室;
【基金】：中央在京高校重大成果转化项目国家科技支撑计划项目(2011AA100903)
【分类号】：Q811.4;TS201.3

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