暹罗芽孢杆菌对多重耐药细菌的抑菌活性及其抗菌次级代谢产物分析  

Analysis of Antibacterial Activity and Antibacterial Secondary Metabolites of Bacillus siamensis Against Multidrug-Resistant Bacteria

作　　者：李海利[1,2] 杨夷平 杨康 雷亚楠 闫祥洲 杨帆[2] 马春江 LI Haili;YANG Yiping;YANG Kang;LEI Yanan;YAN Xiangzhou;YANG Fan;MA Chunjiang(Institute of Animal Husbandry,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;Hami Center for Animal Disease Control and Prevention,Hami 839001,China)

机构地区：[1]河南省农业科学院畜牧研究所,河南郑州450002 [2]哈密市动物疫病预防控制中心,新疆哈密839001

出　　处：《中国兽医杂志》2024年第12期73-80,共8页Chinese Journal of Veterinary Medicine

基　　金：河南省科技攻关项目(242102110053);哈密市科学研究与技术开发计划项目(hmkjjh202311);新疆维吾尔自治区2023年度第二批“2+5”重点人才计划项目(20231058)。

摘　　要：为筛选对多重耐药细菌具有较好抑菌活性的药用植物内生菌,本试验采用平板对峙法和抑菌圈法分离筛选金银花内生菌,通过形态观察、16S rRNA基因PCR扩增和测序对其鉴定,并通过全基因组学分析其抗菌活性物质。结果显示,从金银花根、茎、叶和花中分离、鉴定和筛选出1株抗菌效果较好的革兰阳性内生菌(命名为HNYJ2204B),经鉴定为暹罗芽孢杆菌(Bacillus siamensis)。该菌能有效抑制多重耐药大肠杆菌(E.coli)、胸膜肺炎放线杆菌(APP)、副猪嗜血杆菌(HPS)和巴氏杆菌(PM)。全基因组测序分析结果显示,HNYJ2204B的基因组大小为3929792 bp,G+C含量为46.5%,预测其编码基因个数为3747个。分别有3744、3527、3348、3074、2870和2364个基因被NR、Swiss-Prot、Pfam、COG、GO和KEGG数据库注释,通过antiSMASH软件预测到与次级代谢产物合成相关的8个基因簇;主要次级代谢产物有大环内酯类化合物大环内酰亚胺H、多烯类化合物杆菌烯、脂肽类丰霉素、抗真菌化合物的二肽化合物芽孢菌溶素和多肽类细菌素儿茶酚型杆菌巴汀/嗜铁素类和聚酮类艰难菌素等。从暹罗芽孢杆菌HNYJ2204B的基因组中含有的次级代谢产物可以预测出,该菌具有开发为新药和饲料添加剂的潜力。In order to screen medicinal plant endophytes with good antibacterial activity against multidrug-resistant bacteria,this study employed plate confrontation and inhibition zone methods to isolate and select endophytes from honeysuckle.The isolates were identified through morphological observation,16S rRNA gene PCR amplification,and sequencing,followed by whole-genome analysis of their antimicrobial active substances.The results showed that a Gram-positive endophyte with good antibacterial effects(named HNYJ2204B)was isolated and identified as Bacillus siamensis from the roots,stems,leaves,and flowers of honeysuckle.This strain effectively inhibited multidrug-resistant Escherichia coli,Actinobacillus pleuropneumoniae,Haemophilus parasuis,and Pasteurella multocida.Whole-genome sequencing analysis revealed that the genome size of HNYJ2204B was 3929792 bp,with a G+C content of 46.5%and an estimated number of coding genes of 3747.A total of 3744,3527,3348,3074,2870,and 2364 genes were annotated against the NR,Swiss-Prot,Pfam,COG,GO,and KEGG databases,respectively.The antiSMASH software predicted eight gene clusters associated with secondary metabolite synthesis;the main secondary metabolites included macrolide compound macrolactin H,polyene compound bacillaene,lipopeptide compound fengycin,antifungal compound dipeptide bacilysin,and peptide bacteriocin catechol-type bacillibactin and polyketide difficidin.The secondary metabolites present in the genome of Bacillus siamensis HNYJ2204B indicate its potential for development as a new drug and feed additive.

关 键 词：暹罗芽孢杆菌 金银花 内生菌 抑菌效果 抑抗菌活性物质 

分 类 号：S855.1[农业科学—临床兽医学]