发酵蔬菜来源具抑菌活性明串珠菌的筛选及其细菌素基因簇挖掘  

Screening of Leuconostoc Strains with Antibacterial Activity from Fermented Vegetables and Mining of Bacteriocin Gene Cluster

作　　者：刘毕琴 陈骏飞 罗义勇 赵勇 万幸 蔡英丽 唐蓉 史巧 李宏 LIU Biqin;CHEN Junfei;LUO Yiyong;ZHAO Yong;WAN Xing;CAI Yingli;TANG Rong;SHI Qiao;LI Hong(Institute of Agro-Product Processing,Yunnan Academy of Agricultural Sciences,Kunming 650223,China;National R&D Center for Freshwater Fish Processing,College of Life Sciences,Jiangxi Normal University,Nanchang 330022,China;Faculty of Agriculture and Forestry,University of Helsinki,Helsinki 00790,Finland;College of Food Science and Technology,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南省农业科学院农产品加工研究所,云南昆明650223 [2]江西师范大学生命科学学院国家淡水鱼加工技术研发专业中心,江西南昌330022 [3]赫尔辛基大学农林学院,芬兰赫尔辛基00790 [4]云南农业大学食品科学技术学院,云南昆明650201

出　　处：《食品工业科技》2024年第11期142-150,共9页Science and Technology of Food Industry

基　　金：云南省科技厅重大科技专项(202002AE320006);科技人才与平台计划(202005AD160015)。

摘　　要：为提高发酵蔬菜的安全性和保藏性,从来自云南传统发酵蔬菜的8株明串珠菌中筛选出1株对食源性致病菌和引起泡菜过酸化的细菌抑制效果好的肠膜明串珠菌AP7。通过排除酸和H2O2影响及蛋白酶敏感性确定AP7的主要抑菌物质,分析其酸稳定性和热稳定性,根据AP7的全基因组序列挖掘潜在的细菌素基因簇。结果表明:排除酸及H2O2的影响后,菌株的发酵上清液仍具有明显抑菌活性,经蛋白酶处理后,抑菌效果明显下降,推测AP7发酵上清液浓缩液中的抑菌物质为细菌素;该细菌素对pH变化敏感,热稳定性高,分子质量在6.51~14.4 kDa;全基因组测序表明,菌株AP7全基因组包含1条染色体(1948310 bp)和2个质粒(37366和20698 bp),GC含量37.7%;存在1个以Enterocin_X_chain_beta细菌素为核心的基因簇,其编码产物预测为带正电的亲水性稳定蛋白,二级结构以α-螺旋为主,三级结构主要由两端松散肽链和中间α-螺旋构成。综上,产细菌素的肠膜明串珠菌AP7具有优良抑菌性能,有潜力应用于酸性食品的发酵和防腐。In order to improve the safety and preservability of fermented vegetables,strain Leuconostoc mesenteroides AP7,with potent inhibitory effects on foodborne pathogens and bacteria causing paocai over-acidification,was selected from 8 Leuconostoc strains derived from traditional fermented vegetables in Yunnan,China.The main antimicrobial substance of AP7 was determined by eliminating the effects of organic acid and H2O2 and through protease sensitivity.The acid stability and thermal stability were analyzed and potential bacteriocin gene clusters were mined from the whole genome sequence of AP7.The results showed that the culture supernatant still had obvious antibacterial activity after excluding the influence of acid and H2O2,and after protease treatment,the antibacterial effect decreased significantly.It was speculated that the antibacterial substance in the concentrated culture supernatant was bacteriocin.The bacteriocin was sensitive to pH changes,had high thermal stability,and had a molecular weight of 6.51~14.4 kDa.Whole genome sequencing showed that the whole genome of AP7 contained 1 chromosome(1948310 bp)and 2 plasmids(37366 and 20698 bp),with a GC content of 37.7%.There was a gene cluster with Enterocin_X_chain_beta bacteriocin as the core.Its encoded product was predicted to be a positively charged hydrophilic stable protein,the secondary structure was dominated byα-helix,and the tertiary structure was mainly composed of loose peptide chains at both ends and anα-helix in the middle.In summary,the bacteriocin-producing Leuconostoc mesenteroides AP7 has excellent antibacterial properties and has the potential to be applied to the fermentation and preservation of acidic foods.

关 键 词：发酵蔬菜 肠膜明串珠菌 全基因组测序 细菌素 

分 类 号：TS201.3[轻工技术与工程—食品科学]