枯草芽孢杆菌SNBS-3全基因组测序及其抑菌物质预测分析

Whole Genome Sequencing of Bacillus subtilis SNBS-3 and Prediction of Its Antimicrobial Substances

为了详细解析枯草芽孢杆菌SNBS-3的基因组特征,本研究在前期研究基础上,采用Illumina二代测序技术和第三代高通量Pacbio测序平台,对从传统豆酱中筛选得到的枯草芽孢杆菌SNBS-3进行全基因组测序,获得菌株基因组特征信息、基因功能注释及分类、系统发育进化和次级代谢产物等关键信息。结果表明:SNBS-3的基因组为一条环状闭合DNA,大小为4076387 bp,共有4000个蛋白质编码基因,在直系同源集、基因本体论、京都基因与基因组百科全书、碳水化合物活性酶、抗生素耐药性数据库和致病毒力因子数据库分别注释到3209、2824、2560、147、12个和4个功能基因。应用在线软件AntiSMASH和Bagel4发现其除具有合成Surfactin、Mycosubtilin、Plipastatin、Bacilysin和Bacillaene等多种抑菌物质的相关基因外,还具有一条完整的细菌素Subtilosin A合成基因簇,结合抑菌实验和蛋白酶K实验结果推测枯草芽孢杆菌SNBS-3具有合成细菌素SubtilosinA的能力。综上,枯草芽孢杆菌SNBS-3全基因组测序结果表明其本身可产生多种抑菌物质,是1株具有生防潜力的微生物,相关分析结果可为包括细菌素SubtilosinA在内的多种抑菌物质的进一步开发应用提供理论基础。

As an extension of the previous research,this study aimed to comprehensively characterize the genome of Bacillus subtilis SNBS-3.Illumina second-generation sequencing technology and the third-generation high-throughput Pacbio sequencing platform were used for whole-genome sequencing of B.subtilis SNBS-3 isolated from traditional bean paste to obtain the key information of genome characteristics,gene function annotation and classification,phylogenetic evolution,and secondary metabolites.The results showed that the genome of SNBS-3 was a closed circular DNA of 4076387 bp in length containing 4000 protein-coding genes.A total of 3209,2824,2560,147,12 and 4 functional genes were annotated in the Clusters of Orthologous Groups(COG),Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG),Carbohydrate-Active Enzymes(CAZyme),Comprehensive Antibiotic Resistance Database(CARD)and Virulence Factor Database(VFDB),respectively.Using the online software AntiSMASH and Bagel4,we found that it contained genes related to the synthesis of surfactin,mycosubtilin,plipastatin,bacilysin and bacillaene,as well as a complete gene cluster for the synthesis of the bacteriocin subtilosin A.Based on the results of antimicrobial test and proteinase K test,it was hypothesized that B.subtilis SNBS-3 had the ability to synthesize subtilosin A.In conclusion,the whole genome sequencing results of B.subtilis SNBS-3 show that it can produce a variety of bacteriostatic substances and thus have biocontrol potential.The results from this study provide a theoretical basis for further development and application of various bacteriostatic substances including the bacteriocin subtilosin A.