应用PCR法检测高产胞外多糖双歧杆菌  

Detection of EPS producing bifidobacterium by polymerase chain reaction

作　　者：石莹[1] 呼鑫荣 李静[1] 郭颖[1] 李阳[1] 旭日花[1] 

机构地区：[1]内蒙古大学生命科学学院,内蒙古呼和浩特010021

出　　处：《生物技术世界》2015年第8期1-3,共3页Biotech World

基　　金：内蒙古自治区自然科学基金博士项目(No.2014BS0307);国家自然科学基金面上项目(No.31271827)

摘　　要：胞外多糖(exopolysaccharide,EPS)是细菌分泌到胞外的一种重要的次级代谢产物,筛选高产EPS的菌株对以后其EPS的理论与应用研究具有重要意义。本文采用传统法对分离自婴儿肠道的若干双歧杆菌是否产EPS进行了检测,并探索了高产EPS双歧杆菌的PCR检测法。PCR法中,引物选择双歧杆菌糖基转移酶(Glycosyltransferase family group 2)基因,结果显示这段序列可能与多糖重复单元的合成与装载有关。另外结果还发现,PCR法能检测出产EPS双歧杆菌,扩增片段为1000 bp左右,序列相似度比对结果与各菌株EPS的产量有着明显的相关性,这种方法比传统的测糖产量的方法更省时省力,而且对检测结果的影响因素比较少,结果较准确,是一种值得进行深入研究探讨的新方法。Exopolysaccharide(EPS) is a sort of important secondary metabolite secreted by bacteria. Bifidobacterium has been accepted as an excellent probiotic with many health promoting functions on the host. And EPS produced from Bifidobacterium also have many important functions on humans.Therefore, exploring a fast and accurate way to detect EPS is essential. In this paper, 5 Bifidobacterium strains from healthy baby's faeces were used to screen EPSproducing strains by polymerase chain reaction and detecting EPS yield. The primers were designed according to the Glycosyltransferase family group 2 sequences of Bifidobacterium longum ATCC 15697, and the amplified fragments about 1000 bp were obtained by PCR from each Bifidobacterium strains. Eps O protein which was translated by the fraction was one of the glycosyltransferase enzymes involved in the synthesis of repeating sugar units onto the lipid carrier. By contrasting gene sequences on NCBI, it turned out that the contrast results have an apparent correlation with the yield of EPS detected by phenol-sulfuric acid method. Maybe the PCR is a new method which can detect the yield of exopolysaccharide by using less time, money and machines than traditional methods. We also find it pose a remarkable effect on reducing errors in the EPS yield detecting. Thus, this new method is credible and worthwhile, but deserving deep explorations.

关 键 词：双歧杆菌 胞外多糖 PCR 检测 

分 类 号：TQ929[轻工技术与工程—发酵工程]