产胞外多糖嗜热链球菌eps基因簇的表达及生物信息学分析
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国家自然科学基金地区科学基金项目(31860448)


Expression of eps Gene Cluster in Streptococcus thermophilus Exopolysaccharide and Bioinformatics Analysis
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    目的:胞外多糖(EPS)作为乳酸菌(LAB)重要的次级代谢产物,在发酵乳制品的生产中发挥着重要作用。采用生物信息学方法分析乳酸菌产EPS的分子机制,为开发利用EPS提供理论依据。方法:采用二代测序技术(Illumina Hiseq 4000)及实时荧光定量PCR技术(RT-qPCR)对分离自蒙古国酸牛奶中一株高产EPS的嗜热链球菌IMAU20756进行基因组测序,并对不同时间点eps基因表达量进行定量分析。结果:该菌株基因组全长为1 838 440 bp,GC含量为38.8%,共编码2 120个基因,包含1 962个功能基因,36个tRNA、515个假基因、1个tmRNA和2个重复单位。其中和EPS生产相关的基因有12个,epsA、epsB是调控基因,epsC、epsD决定多糖链长,eps1E、eps2E、epsG、epsF、epsH、epsI、epsJ是编码糖基转移酶基因,epsK调控多糖聚合及输出。结论:所有和EPS生物合成相关的基因在发酵过程中均能表达,特别是糖基转移酶的基因表达量在发酵6 h时达最高。

    Abstract:

    Objective: Extracellular polysaccharides(EPS), as important secondary metabolites of lactic acid bacteria (LAB) play an important role in the production of fermented dairy products. Therefore, it is necessary to analyze the molecular mechanism of EPS production by lactic acid bacteria by bioinformatics in order to provide theoretical basis for the development and utilization of EPS. Methods:In this experiment, the second generation sequencing technology (Illumina Hiseq 4000) and Real-time fluorescent quantitative PCR(RT-qPCR) were used to sequence the genome of a strain of Streptococcus thermophilus IMAU20756 with high production of EPS from Mongolian yoghurt and quantitative analysis of eps gene expression at different time points. Results: The total genome length of the strain was 1 838 440 bp and GC contained 38.8%. In addition, it encoded 2 120 genes, including 1 962 functional genes, 36 tRNA, 515 pseudogenes, 1 tmRNA and 2 repetitive units. Among them, there are 12 genes related to EPS production. EpsA and epsB are regulatory genes, epsC and epsD determine the chain length of polysaccharides, eps1E, eps2E, epsG, epsF, epsH, epsI and epsJ are genes that encode glycosyltransferase, and epsK regulates the polymerization and output of polysaccharides. Conclusion: All the genes related to EPS biosynthesis could be expressed during fermentation, especially the gene expression of glycosyltransferase was the highest at 6 h of fermentation.

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田佳乐;刘洋;李嘉雯;乔少婷;胡海敏;丹彤.产胞外多糖嗜热链球菌eps基因簇的表达及生物信息学分析[J].中国食品学报,2021,21(11):244-251

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  • 在线发布日期: 2021-12-17
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