摘要：
High performance liquid chromatography (HPLC) was used to analyze the contents of organic acids and its changing trend during Chinese rice wine fermentation.To identify acid-producing bacteria in Chinese rice wine fermentation broth,a method unified the modern molecular biology and the traditional microbiology was used.The analysis on fermentation characteristics was also carried out.The results indicated that lactic acid,acetic acid and citric acid were the major organic acids accounting for more than 55％ of the content of total organic acids.The content of organic acids showed a upward trend along with fermentation process,wherein citric acid,oxalic acid,tartaric acid,lactic acid,malic acid,pyruvic acid and acetic acid increased largely,while succinic acid,fumaric acid,α-ketoglutaric acid did not increased significantly.The results from denatured gradient gel electrophoresis (DGGE) showed that it contained Lactobacillus,Saccharopolyspora,Staphylococcus,Bacillus licheniformis,Streptomyces sp.and Uncultured bacterium during fermentation.The acid-producing bacteria identified by comparison on 16S rDNA sequence and fermentation characteristics were Lactobacillus,especially Lactobacillus plantarum,which belonged to heterofermentation lactic acid bacteria.%采用高效液相色谱法(high performance liquid chromatography,HPLC)分析了绍兴黄酒发酵过程中有机酸组成及动态变化情况,运用变性梯度凝胶电泳技术(denatured gradient gel electrophoresis,DGGE)分析了细菌群落结构变化,以纯培养结合分子生物学的方法分离、鉴定了产酸细菌,并对其发酵特性进行了研究.结果表明,乳酸、乙酸和柠檬酸是黄酒发酵过程中产生的主要有机酸,其含量占总有机酸的55％以上.有机酸的含量随发酵的进行呈递增趋势,柠檬酸、草酸、酒石酸、乳酸、苹果酸、丙酮酸和乙酸含量增幅较大(90％ ～385％),琥珀酸、富马酸、α-酮戊二酸增幅较小(28％ ～34％).DGGE分离及测序结果表明,黄酒发酵过程中主要的细菌有乳酸杆菌(Lactobacillus)、糖多孢菌(Saccharopolyspora)、葡萄球菌(Staphylococcus)、地衣芽孢杆菌(Bacillus lichenifor-mis)、链霉菌(Streptomyces sp.)以及不可培养细菌(Uncultured bacterium)等.经16S rDNA序列分析鉴定及发酵特性研究,产酸细菌主要为乳酸杆菌,其中植物乳杆菌占70％,属于异型发酵乳酸菌.
