氨肽酶生产菌株Bacillus subtilis LBJ4-5的全基因组测序及基因功能分析  

Whole-genome Sequencing and Sequence Analysis of an Aminopeptidase-producing Bacillus subtilis LBJ4-5

作　　者：郭宝松 张玉姣 代艺伟 陈映羲 董亮[1] 张素芳 GUO Baosong;ZHANG Yujiao;DAI Yiwei;CHEN Yingxi;DONG Liang;ZHANG Sufang(Collaborative Innovation Center of Seafood Deep Processing,National Engineering Research Center of Seafood,SKL of Marine Food Processing&Safety Control,School of Food Science and Technology,Dalian Polytechnic University,Dalian 116034,China)

机构地区：[1]大连工业大学食品学院,海洋食品加工与安全控制全国重点实验室,国家海洋食品工程技术研究中心,海洋食品精深加工关键技术省部共建协同创新中心,辽宁大连116034

出　　处：《食品工业科技》2024年第24期32-41,共10页Science and Technology of Food Industry

基　　金：国家自然科学基金(32072185);山西省重点研究计划(202202130501011)。

摘　　要：本研究从郫县豆瓣中分离获得一株高产氨肽酶菌株B. subtilis LBJ4-5,通过溶血性试验及抗生素敏感性试验评估了其安全性。同时,采用Illumina二代测序技术和第三代高通量Pacbio测序平台对B. subtilis LBJ4-5进行全基因组测序,获得其基因组特征信息,并通过COG、GO、KEGG、CAZY、CARD和VFDB等数据库进行基因预测及功能注释,以及评价其氨肽酶生产性能和安全性。结果显示,B. subtilis LBJ4-5经48 h摇瓶培养后发酵上清中的氨肽酶活达到179.23 U/mL,菌株无溶血活性且对头孢唑林、万古霉素、青霉素G、头孢氨苄等10种抗生素均高度敏感(直径≥20 mm),显示出良好的安全性。B. subtilis LBJ4-5的基因组由一条环状闭合DNA及一个环状质粒组成,大小为4053926 bp,GC含量为43.66%,共预测到4285个蛋白质编码基因、85个tRNA基因、33个rRNA基因、119个sRNA。在COG、GO、KEGG、CAZY、CARD和VFDB数据库中分别注释到3386、2581、2552、144、256和401个功能基因。进一步的信息挖掘分析表明,B. subtilis LBJ4-5的基因组中含有31个氨肽酶编码基因。而毒力基因与耐药性基因的预测分析,进一步证实了B. subtilis LBJ4-5菌株的安全性。综上,B.subtilis LBJ4-5可用作发酵食品加工的发酵菌剂。In this study,a high-yield aminopeptidase producing strain,Bacillus subtilis LBJ4-5,was isolated from Pixian Douban,and its safety was evaluated through hemolysis and antibiotic sensitivity tests.Additionally,Illumina second-generation sequencing and PacBio third-generation high-throughput sequencing were employed for the whole-genome sequencing of B.subtilis LBJ4-5 to characterize its genomic features.Gene prediction and functional annotation were carried out using databases such as COG,GO,KEGG,CAZY,CARD and VFDB.Furthermore,the aminopeptidase production capacity and safety of the strain were evaluated.The results showed that after 48 hours of shake flask cultivation,the aminopeptidase activity in the fermentation supernatant of B.subtilis LBJ4-5 reached 179.23 U/mL.The strain showed no hemolytic activity and was highly sensitive(diameter≥20 mm)to 10 antibiotics,including cefazolin,vancomycin,penicillin G,and cefadroxil,indicating a favorable safety profile.The genome of B.subtilis LBJ4-5 consisted of a circular closed DNA and a circular plasmid,with a size of 4053926 bp and a GC content of 43.66%.A total of 4285 protein-coding genes,85 tRNA genes,33 rRNA genes,and 119 sRNAs were predicted.Functional gene annotations in COG,GO,KEGG,CAZY,CARD,and VFDB databases identified 3386,2581,2552,144,256 and 401 functional genes,respectively.Further analysis indicated that the genome of B.subtilis LBJ4-5 contains 31 aminopeptidase-encoding genes.Additionally,the prediction of virulence and antibiotic resistance genes further confirmed the safety of the B.subtilis LBJ4-5 strain.In conclusion,B.subtilis LBJ4-5 can be used as a fermentation starter for the processing of fermented foods.

关 键 词：枯草芽孢杆菌 全基因组测序 氨肽酶 基因功能注释 郫县豆瓣 

分 类 号：TS201.3[轻工技术与工程—食品科学]