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以猪链球菌的细菌素为研究目标,采用琼脂扩散法筛选出27株产类细菌素猪链球菌菌株,其中CZJYW024菌株的细菌素抑菌活性最强。使用细菌素检测引物(nisin A和scn A)对产类细菌素菌株进行检测,scn A和nisin A的携带率分别为33.3%和14.8%。初步纯化CZJYW024菌株的抑菌物质,提取物对马链球菌兽疫亚种ATCC35246的抑菌环直径达(23.30±0.20)mm,经有机酸、过氧化氢干扰试验及质谱鉴定,证实该抑菌物质属于羊毛硫细菌素家族,对热、pH和蛋白酶稳定,命名为suicin024。抑菌试验证实该基因缺失株抑菌活性消失,进一步分析发现suicin024包含10个开放阅读框,由scz基因簇编码。CZJYW024菌株经BALB/c小鼠模型证实为无毒株。本研究为该细菌素在动物制药和食品原料加工等抗生素替代品的研究和开发上提供理论依据。
Abstract：Bacteriocin is a kind of antimicrobial secretory proteins produced by a variety of microotganisms. In this study,27 of 530 strains of Streptococcus suis exhibited antibacterial activity against pathogenic Gram-positive isolates. Two bacteriocin-related structural genes( scn A and nisin A) in these 27 strains were detected by PCR,showing the prevalence of scn A and nisin A with 33. 3% and 14. 8%,respectively. S.suis CZJYW024 exhibited the strongest antibacterial activities. Then the inhibitory substances produced by this strain were purified,whose inhibitory zone was up to 23. 30±0. 20 mm against Streptococcus equi ssp. zooepidemicus ATCC35246. After eliminating the interference of organic acids and hydrogen peroxide,the modified amino acids sequencing of the purified bacteriocin obtained by Q-Exactive suggested a lantibiotic. The bacteriocin,named suicin024,was heat,pH and protease resistant. The putative gene cluster involved in suicin024 produced by S. Suis CZJYW024 was amplified by PCR and sequence analysis revealed the presence of 10 open reading frames named scz gene cluster. To assess the contribution of the gene cluster to antibacterial activity,the crucial scz cluster was deleted to generate the mutant strain △scz. The△scz mutant strain lost the ability to produce bacteriocin compared with its parental strain,suggesting that suicin024 was encoded by scz gene cluster. Given that S. suis CZJYW024 can not cause infections in a mouse model,the bacteriocin produced by this bacterial strain is regarded as a potential alternative for conventional antibiotics,with promising applications in both the food and pharmaceutical industries.
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基本信息:
DOI：
中图分类号:S852.611
引用信息:
[1]马烨,张越,姚火春.产细菌素猪链球菌的筛选及其基因簇功能分析[J].畜牧与兽医,2017,49(05):89-97.
基金信息:
农业部行业公益性专项项目(201303041)