本文采用16S rDNA PCR- DGGE（变性梯度凝胶电泳）指纹图谱和系统发育分析方法，以高盐稀醪发酵工艺生产的广东酱油为研究对象，揭示了酱油生产发酵过程中细菌群落结构的多样性及动态变化。从样品中提取总细菌DNA，用降落PCR扩增16S rDNA V3片段序列，再通过分析DGGE图谱选择特异性条带，进行割胶回收、测序及Blast分析。DGGE图谱表明，发酵初期样品具有丰富的微生物群落，但之后只有少数种类细菌存活，整个酱油发酵过程微生物群落结构的演变规律是由复杂到简单，这也说明酱油发酵环境具有抑制微生物生长的作用。测序结果表明，代表最相似菌为魏斯菌属（Weissella cibaria）和非培养的肠杆菌属（Uncultured Enterobacter sp.），其次是嗜盐四联球菌（Tetragenococcus halophilus）、类肠膜魏斯氏菌（Weissella paramesenteroides）、弗氏柠檬酸杆菌（Citrobacter freundii）、产气肠杆菌（Enterobacter aerogenes）和肠杆菌属（Enterobacter sp. BF1-8）。

Techniques such as 16S rDNA polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting and phylogenetic analysis were used to reveal the dynamic changes in bacterial community structure during a high-salt-diluted fermentation of guangdong soy sauce. Total bacterial DNA was extracted from the samples and touchdown PCR was performed to amplify the V3 region of 16S rDNA. Subsequently, based on DGGE fingerprints, the specific gel bands were excised, sequenced, and analyzed using Basic Local Alignment Search Tool (BLAST). The DGGE fingerprints showed that various types of bacterial communities existed in the initial fermentation period, but only few survived in the later stages. The evolutionary pattern of the microbial community structure during soy sauce fermentation progressed from complex to simple, which demonstrated that the growth of microorganisms was inhibited by the conditions of fermentation. The sequencing identified the bacterial strains as Weissella cibaria and uncultured Enterobacter sp. with highest similarity, followed by Tetragenococcus halophilus, Weissella paramesenteroides, Citrobacter freundii, Enterobacter aerogenes, and Enterobacter sp. BF1-8.

广东省科技计划项目（2010B020316004）