一种基于培养技术的豆瓣酱合成微生物群落结构检测方法的建立及应用  

Development and application of a culture-dependent method for quantifying composition of a synthetic broad bean paste microbial community

作　　者：易峰 刘春凤[1] 李宛格 钮成拓 李崎[1] 郑飞云[1] 王金晶 YI Feng;LIU Chunfeng;LI Wange;NIU Chengtuo;LI Qi;ZHENG Feiyun;WANG Jinjing(Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China;School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]工业生物技术教育部重点实验室(江南大学),江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与发酵工业》2024年第24期304-312,330,I0001,共11页Food and Fermentation Industries

基　　金：国家自然科学基金项目(32272282)。

摘　　要：该研究旨在通过药敏试验筛选出具有选择性的抗生素组合,以建立能够量化豆瓣酱合成微生物群落中任一菌株绝对丰度的选择性培养基。采用微量稀释法分析了42种抗生素对肉葡萄球菌、枯草芽孢杆菌、解淀粉芽孢杆菌、嗜盐四联球菌和融合魏斯氏菌的抑菌活性,并筛选出能通过培养将这些特定细菌有效区分开的抗生素。通过组合搭配能抑制细菌的氨苄西林钠、萘夫西林钠、盐酸金霉素、恶喹酸和林可霉素,以及能抑制真菌的制霉菌素,设计了一系列选择性培养基。在仅接种目标菌株与接种混合菌悬液的选择性平板上,目标菌株的菌落总数无显著差异(P>0.05)。这一系列选择性培养基能够精确量化由鲁氏接合酵母、肉葡萄球菌、枯草芽孢杆菌、解淀粉芽孢杆菌、嗜盐四联球菌和融合魏斯氏菌组成的合成微生物群落中任一菌株的绝对丰度。通过应用该方法,成功分析了豆瓣酱合成微生物群落的结构以及群落成员间的相互作用。该研究为成员数量较少的合成微生物群落菌群结构检测方法的建立提供了一种简便、准确、低成本的解决方案。This study aimed to screen for selective antibiotic combinations through antimicrobial susceptibility tests,establishing selective media that can quantify the absolute abundance of any strain in a synthetic broad bean paste microbial community.The antimicrobial activity of 42 antibiotics against Staphylococcus carnosus,Bacillus subtilis,Bacillus amyloliquefaciens,Tetragenococcus halophilus,and Weissella confusa was analyzed using microdilution method,and antibiotics that could effectively distinguish these specific bacteria through cultivation were screened.Selective media were designed by combining ampicillin sodium,nafcillin sodium,chlortetracycline hydrochloride,oxolinic acid,lincomycin,and antifungal nystatin.The CFU number of the target strain on selective plates inoculated with the target strain and mixed microbial suspension did not show a significant difference(P>0.05).These selective media could accurately quantify the absolute abundance of any strain in the synthetic microbial community composed of Zygosaccharomyces rouxii,Staphylococcus carnosus,Bacillus subtilis,Bacillus amyloliquefaciens,Tetragenococcus halophilus,and Weissella confusa.By applying this method,the composition of a synthetic broad bean paste microbial community and the interactions between community members were successfully analyzed.This study provides a simple,accurate and low-cost solution for the development of methods for quantifying the composition of synthetic microbial communities with fewer members.

关 键 词：豆瓣酱 合成微生物群落 抗生素 抑菌率 微生物群落结构检测方法 

分 类 号：TS264.24[轻工技术与工程—发酵工程]