产细菌素贝莱斯芽孢杆菌CM7-4全基因组测序及抑菌活性研究

doi:10.16431/j.cnki.1671-7236.2024.12.001

摘要/Abstract

摘要： 【目的】贝莱斯芽孢杆菌(Bacillus velezensis)CM7-4是从海水中分离的具有良好抗菌活性的芽孢杆菌。本试验旨在探究贝莱斯芽孢杆菌CM7-4的抗菌机理，并对其全基因组序列进行分析，以期挖掘其抗菌等功能特性基因。【方法】通过酸沉淀法得到贝莱斯芽孢杆菌CM7-4的细菌素粗提物，通过固体琼脂打孔法测定蛋白酶K对其抑菌效果的影响；基于MGI DNBSEQ-T7平台对贝莱斯芽孢杆菌CM7-4进行全基因组测序分析，通过序列组装、基因预测和功能注释分析其产细菌素的潜质。【结果】贝莱斯芽孢杆菌CM7-4的细菌素粗提物对大肠杆菌、沙门菌、粪肠球菌、单增李斯特菌等8种不同的革兰阴性和阳性菌具有良好的抑菌效果；与对照组相比，经蛋白酶K处理后细菌素粗提物对8种指示菌的抑菌效果均极显著下降(P＜0.01)；全基因组测序显示，贝莱斯芽孢杆菌CM7-4的基因组大小为3 953 124 bp，GC含量为47.04%；共预测到3 870个蛋白编码基因、9个rRNA、81个tRNA、1个ncRNA；基因组中含有11个与细菌素合成相关的基因、10个与抗氧化活性相关基因，以及4个与调控免疫和炎症信号通路相关的基因。【结论】贝莱斯芽孢杆菌CM7-4是一株潜在的产细菌素菌株，为进一步研究其抗菌特性提供了理论基础。

关键词: 芽孢杆菌; 全基因组测序; 细菌素; 基因功能注释

Abstract: 【Objective】 Bacillus velezensis CM7-4 was isolated from seawater and had been demonstrated to possess good antibacterial activity.This study was to investigate the antimicrobial mechanism of Bacillus velezensis CM7-4 and analyze the whole genome sequence in order to explore its antibacterial and other functional genes.【Method】 The bacteriocin crude extract of Bacillus velezensis CM7-4 was obtained by acid precipitation,and the effect of proteinase K on its antibacterial effect was determined by the solid agar perforation method.Subsequently,the complete genome of Bacillus velezensis CM7-4 was sequenced on the MGI DNBSEQ-T7 platform.Their bacteriocin production potential was analyzed by sequence assembly,gene prediction and functional annotation.【Result】 The bacteriocin crude extract of Bacillus velezensis CM7-4 demonstrated a notable bacteriostatic effect on a diverse range of 8 kinds of Gram-negative and positive bacteria,including Escherichia coli,Salmonella,Enterococcus faecalis,Listeria monocytogenes and others. Compared with control group, the bacteriocin crude extract demonstrated a extremely significantly diminished inhibitory effect on the eight indicator bacteria following treatment with proteinase K (P＜0.01).Whole genome sequencing revealed that the genome size of Bacillus velezensis CM7-4 was 3 953 124 bp with a 47.04% GC content.A total of 3 870 protein-coding genes,9 rRNA, 81 tRNA and 1 ncRNA were identified.And 11 bacteriocin synthesis genes,10 antioxidant activity genes,and 4 immune and inflammatory signalling pathway regulation genes were predicted.【Conclusion】 The results of this experiment indicated that Bacillus velezensis CM7-4 was a potential bacteriocin-producing strain.This study provided a theoretical basis for further study of its antibacterial properties.

Key words: Bacillus; whole genome sequencing; bacteriocin; gene function annotation

中图分类号:

S816.7

马浩天, 李杨, 余梦博, 潘瑞雪, 龙赟而, 赵一宁, 彭金菊, 马驿. 产细菌素贝莱斯芽孢杆菌CM7-4全基因组测序及抑菌活性研究[J]. 中国畜牧兽医, 2024, 51(12): 5117-5127.

MA Haotian, LI Yang, YU Mengbo, PAN Ruixue, LONG Yuner, ZHAO Yining, PENG Jinju, MA Yi. Whole Genome Sequencing of Bacteriocin-producing Bacillus velezensis CM7-4 and Its Bacteriostatic Activity[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(12): 5117-5127.

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