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【目的】筛选一株可转化大豆苷元为S-雌马酚的微生物菌株,并对该菌株进行鉴定。【方法】在厌氧条件下采用抗生素抑制非目标菌生长并结合稀释涂平板法进行菌株分离,分离可转化大豆苷元生成S-雌马酚的肠道细菌,并对产物进行结构鉴定。之后通过16S rDNA序列分析,构建该菌系统进化树,结合菌体形态及菌落特征,确立该菌系统发育学地位。【结果】从大鼠肠道内筛选分离到一株可以将大豆苷元转化为S-雌马酚的革兰氏阴性兼性厌氧菌株LH-52(JN861767),16S rDNA序列测序结果 BLAST比对表明该菌株与奇异变形杆菌(Proteus mirabilis)相似度达到了99%,结合形态特征和生理生化实验结果鉴定该菌为奇异变形杆菌。根据HPLC保留时间、质谱、核磁共振等波谱数据分析确定产物为S-雌马酚。【结论】菌株P.mirabilis LH-52为首次筛选到的可转化大豆苷元为S-雌马酚的兼性厌氧菌,相对于文献报道的严格厌氧菌更适合于工业化生产。
【Objective】 A strain of microorganism that can transform daidzein to S-equol was screened and the strain was identified. 【Method】 Antibacterial agents were used to inhibit the growth of non-target bacteria under anaerobic conditions. The bacteria were isolated by dilution plate method. The intestinal bacteria that could transform daidzein to produce S-equol were isolated and their structures were identified. After 16S rDNA sequence analysis, the phylogenetic tree of the bacteria was constructed, and the morphological and colony characteristics of the bacteria were combined to establish the phylogenetic position of the bacteria. 【Result】 A Gram-negative facultative anaerobic strain LH-52 (JN861767) that could convert daidzein to S-equol was isolated from the intestine of rats. The BLAST results of 16S rDNA sequencing The results showed that the similarity of the strain with Proteus mirabilis reached 99%. The morphological and physiological and biochemical experiments showed that the strain was Proteus mirabilis. According to HPLC retention time, mass spectrometry, nuclear magnetic resonance and other spectroscopic data analysis to determine the product is S-equol. 【Conclusion】 The strain P. mirabilis LH-52 was the facultative anaerobic bacterium that could be transformed into S-equol for the first time. Compared with the strict anaerobic bacteria reported in the literature, it is more suitable for industrial production.