抗猪链球菌植物乳杆菌细菌素生物学特性及其编码基因簇分析  

Biological Characterization of Bacteriocins of Lactiplantibacillus plantarum against Streptococcus suis and Analysis of Their Encoding Gene Clusters

作　　者：邹鸣晓 王云飞 苏惠 李涵诺 许诺 李国江 董文龙 ZOU Mingxiao;WANG Yunfei;SU Hui;LI Hannuo;XU Nuo;LI Guojiang;DONG Wenlong(College of Animal Science and Technology,Jilin Agricultural Science and Technology University,Jilin Jilin 132109,China;College of Animal Science and Technology,Jilin Agricultural University,Jilin Changchun 130118,China;Jilin Province Key Laboratory of Preventive Veterinary Medicine,Jilin Jilin 132109,China;Jilin Province Technology Innovation Center of Pig Ecological Breeding and Disease Prevention and Control,Jilin Jilin 132109,China)

机构地区：[1]吉林农业科技学院动物科技学院,吉林吉林132109 [2]吉林农业大学动物科学技术学院,吉林长春130118 [3]吉林省预防兽医学重点实验室,吉林吉林132109 [4]吉林省猪生态养殖及疫病防控科技创新中心,吉林吉林132109

出　　处：《饲料工业》2025年第6期101-108,共8页Feed Industry

基　　金：吉林省科技创新平台补助项目[YDZJ202302CXJD038]。

摘　　要：研究基于猪链球菌(Streptococcus suis,S.suis)耐药性加剧现状,分析植物乳杆菌细菌素抗菌生物学特性及其编码基因簇,旨在探究植物乳杆菌细菌素作为猪链球菌替抗产品的潜力。通过16S rRNA基因测序方法对分离获得的乳酸菌菌株进行鉴定,建立并分析菌株生长曲线及其发酵上清液抑菌动力学曲线,确定菌株最佳培养时间,并测定发酵上清液蛋白酶稳定性、热稳定性、酸碱稳定性及紫外线稳定性。通过全基因组测序技术分析菌株基因组信息及细菌素编码基因簇。结果显示:分离得到植物乳杆菌(Lactiplantibacillus plantarum,L.plantarum)一株(W-1),其发酵上清液对猪链球菌有较好的抗菌活性。植物乳杆菌W-1的对数生长期为发酵4~6 h,24 h内其最佳培养时间约为18 h,此时其发酵上清液抗菌活性最强。植物乳杆菌W-1发酵上清液对胰蛋白酶敏感性较高,推测其中产生的抗菌活性的物质细菌素。该细菌素具有优异的热稳定性、酸碱稳定性及紫外线稳定性,于40~100℃、pH 2.0~10.0、紫外线处理30~120 min后均能保持较强的抗菌活性。全基因组测序结果表明,植物乳杆菌W-1染色体基因组大小为3297581 bp,GC占比44.47%。细菌素编码基因簇结果中,由结构基因编码参与合成ClassⅡb类细菌素PlnJK、PlnEF的核心肽起重要抗菌作用。植物乳杆菌W-1所产植物乳杆菌素对猪链球菌具有良好且稳定的抗菌效果,有作为猪链球菌替抗品的潜质。The aim of this study was to analyze the antimicrobial biological characterization of Lactiplantiba-cillus plantarum(L.plantarum)bacteriocin and its coding gene cluster in light of the escalating drug resis-tance in Streptococcus suis(S.suis),with the objective of investigating its potential as an antibiotic replace-ment product for S.suis.The 16S rRNA sequenc-ing method was used to identify the isolated Lacto-bacillus strains,to establish and analyze the growth curves of the strains and the bacteriostatic kinetic curves of their fermentation supernatants,to deter-mine the optimal incubation time of the strains,and to measure the protease stability,thermal sta-bility,acid-base stability,and ultraviolet stability of the fermentation supernatants.The genomic informa-tion of the strain and the bacteriocin coding gene cluster were analyzed by whole-genome sequencing.The results showed that a strain of L.plantarum(W-1)was isolated and its fermentation supernatant showed good antibacterial activity against S.suis.The logarithmic growth phase of L.plantarum W-1 occurs between 4 to 6 h of fermentation,with the optimal cultivation time being approximately 18 h within a 24 h period,at which point the antibacterial activity of its fermentation supernatant is the strongest.The fermenta-tion supernatant of L.plantarum W-1 was more sensitive to trypsin,so it was hypothesized that the sub-stance producing antibacterial activity in it was bacteriocin.The bacteriocin had excellent thermal,acid-base and UV stability,and maintained strong antibacterial activity after 40-100℃,pH 2.0-10.0 and UV treatment for 30-120 min.Whole-genome sequencing results indicate that the chromosome genome size of L.plantarum W-1 is 3297581 bp,with a GC content of 44.47%.In the results pertaining to the bacteriocin-encoding gene cluster,core peptides encoded by structural genes that are involved in the syn-thesis of ClassⅡb bacteriocins Pln JK and Pln EF play a significant role in antimicrobial activity.The plan-taricin produced by L.plantarum W-1 h

关 键 词：细菌素 猪链球菌 全基因组测序 基因簇 植物乳杆菌 

分 类 号：S816.7[农业科学—饲料科学]