克隆文库分析镇江香醋发酵过程中乙偶姻合成基因的多样性  

Clone library analysis of enzyme genes in metabolic pathway for acetoin production in the fermentation of Zhenjiang aromatic vinegar

作　　者：刘娜[1] 陆震鸣[1] 王丽娟[1] 余永健[4] 李国权[4] 史劲松[1,2] 许正宏[1,2,3] 

机构地区：[1]江南大学药学院,工业生物技术教育部重点实验室,无锡214122 [2]国家固态酿造工程技术研究中心,泸州646000 [3]中国科学院天津工业生物技术研究所,天津市工业生物系统与过程工程重点实验室,天津300308 [4]江苏恒顺醋业股份有限公司,镇江212043

出　　处：《应用与环境生物学报》2017年第1期134-139,共6页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(31271922,31530 055);国家高技术研究发展计划(863计划)项目(2012AA021301,2013AA102106,2014A A021501)资助~~

摘　　要：乙偶姻是传统发酵食醋重要的风味物质,也是食醋中活性物质川芎嗪形成的重要前体.采用克隆文库分析方法对镇江香醋醋醅中乙偶姻合成途径的两个关键酶(乙酰乳酸合成酶ALS,乙酰乳酸脱羧酶ALDC)的基因多样性进行分析.结果表明,ALS基因克隆文库中相对丰度最大为Lactobacillus buchneri(65.7%),其次为Acetobacter pasteurianus(21.3%)、L.reuteri(10.2%)、L.casei(1.9%)、L.brevis(0.9%);而ALDC基因克隆文库中A.pasteurianus(91.2%)占绝对优势,且呈现丰富的种内基因多样性,另外还有少量L.fermentum(8.8%).因此,Acetobacter和Lactobacillus可能是醋醅微生物群落中利用丙酮酸合成乙偶姻的主要参与者;本研究可为解析多菌种醋酸发酵过程中乙偶姻微生物合成机理奠定研究基础,同时为其他微生物群落功能的研究提供参考.This research aimed to study the gene diversity of two key enzymes in the acetoin pathway in the vinegar Pei and to lay foundation for the functional microbial community reconstruction. Degenerate primers were designed, validated and used for the amplification of gene coding for acetolactate synthase（ALS） and acetolactate decarboxylase（ALDC） in vinegar Pei. Then two gene clone libraries were established respectively and analyzed by rarefaction curve, cluster analysis and phylogenetic analysis. Two pairs of degenerate primers were validated to be of high specificity and used for the clone libraries construction. Sequences in ALS clone library were clustered to 6 Operational Taxonomic Units（OTUs） and the most abundant species was proved to be Lactobacillus buchneri（65.74%）. Sequences in ALDC clone library were clustered to 16 OTUs and the most abundant species was Acetobacter pasteurianus（91.23%）. The research found that Lactobacillus and Acetobacter might be the main functional genera for the production of ALS and ALDC in the vinegar Pei.

关 键 词：镇江香醋 乙偶姻 基因多样性 克隆文库 微生物群落 乙酰乳酸合成酶 乙酰乳酸脱羧酶 

分 类 号：TS264.22[轻工技术与工程—发酵工程]