基于宏基因组测序和纯培养技术解析胀袋生抽酱油的微生物类群

Analysis of microbial groups in aerogenesis soy sauce based on metagenome sequencing and pure culture technology

该研究采用宏基因组测序技术对胀袋生抽酱油微生物类群进行了解析,对其微生物菌株进行了分离鉴定,并对分离株是否可导致酱油胀气进行了验证。宏基因组结果表明,胀袋生抽酱油样品中平均相对含量大于1.0%的微生物主要由嗜盐四联球菌(Tetragenococcus halophilus)、酸鱼联合乳杆菌(Ligilactobacillus acidipiscis)、植物乳植物杆菌(Lactiplantibacillus plantarum)、耐酸乳杆菌(Lactobacillus acetotolerans)、棒状腐败乳杆菌(Loigolactobacillus coryniformis)、短促生乳杆菌(Levilactobacillus brevis)和屎肠球菌(Enterococcus faecium)构成,平均相对含量分别为28.01%、25.50%、9.45%、2.49%、1.83%、1.53%和1.33%;采用纯培养技术,分离出的49株细菌被鉴定为6个属下的11个种,其中枯草芽孢杆菌(Bacillus subtilis)、破布子联合乳杆菌(Ligilactobacillus pobuzihii)、蛋白原酶腐败乳杆菌(Loigolactobacillus rennini)和L.acidipiscis分离株占总分离株数的39.22%、19.61%、11.76%和9.80%,分离出的2株酵母菌均被鉴定为扣囊复膜酵母(Saccharomycopsis fibuligera);产气验证实验发现,所有分离株均不能单独引起生抽酱油的胀袋现象。由此可见,胀袋生抽酱油具有较高的微生物多样性,在后续研究引入培养组学策略以获得更多的分离株,亦或考虑多菌株的协同作用,对酱油涨袋这一产业问题的解决可能具有积极的意义。

In this study,the metagenome sequencing technology was used to analyze the microbial community of aerogenesis soy sauce,isolate and identify its microbial strains,and verify whether the isolated strains can cause the swelling of the soy sauce.The results of metagenomics showed that the microorganisms with an average relative content of more than 1.0%were mainly composed of Tetragenococcus halophilus,Ligilactobacillus acidipiscis,Lactiplantibacillus plantarum,Lactobacillus acetotolerans,Loigolactobacillus coryniformis,Levilactobacillus brevis and Enterococcus faecium,with average relative contents of 28.01%,25.50%,9.45%,2.49%,1.83%,1.53%and 1.33%respectively;Using pure culture technology,49 bacterial strains isolated were identified as 11 species under 6 genera,including Bacillus subtilis,Ligilactobacillus pobuzihii,Loigolactobacillus rennini and L.acidipiscis accounted for 39.22%,19.61%,11.76%and 9.80%of the total isolated strains,and two strains of yeast were identified as Saccharomyces cerevisiae.The gas production verification experiments revealed that all isolates could not cause the aerogenesis phenomenon of soy sauce alone.It is evident that aerogenesis soy sauce has a high microbial diversity,and the introduction of a culture-omics strategy to obtain more isolates or the consideration of synergistic effects of multiple strains in subsequent studies may have positive implications for the solution of the industrial problem of aerogenesis soy sauce.