The Specified Identification Of New Species In The Pit Mud Of Chinese Liquors And The Research On The Metabolism Of Carbon Sources

As one of the world sixdistilled liquor, Chinese liquorhas a long history and has been favored by people in daily lives. The strong aroma style Chinese liquor shares about 60% of Chinese wine marketin the 2014. The bacteria in the cellar mud produced important fragrance substance or flavor producors, such as acid, alcohol or esterthat, which are important for the liquor flavor. Therefore, the main research method in this thesis is to separate advantagebacteria of 10 years old pit muds. Two new species of bacteria isolated from the cellar mud were analyzed in morphological, physiological, biochemical and molecular biology. The carbon sourcemetabolic diversity of different kinds of isolated strains was analyzed followed by Biolog carbon analyzer system. Fermentation experiment was carried out using the selected strains that showedstrong metabolic intensity. The forms and composition of metabolic of isolated strains were detectedby GC-MS.The results showed that 120 bacteria isolated form 10 years old cellar mud were divided into seven genera:Bacillus, Brevibacillus, Clostridium, Domibacillus, Lactobacillus, Rummellibacillus and Sporolactobacillus. Clostridium, Bacillus and Lactobacillus were primary genera and accounted about 75.8% of all isolates. In addition, most bacteria were isolated by anaerobic way. In these strains, there are two new species of bacteria were named by FW431 and FW731. The 16S rRNA similarity of the two strains were highest closed to the type strainClostridium ljungdahlii DSM 13528T, similarities showed 95.0% and 94.9% respectively. Microscopy assay showed that strains FW431 and FW731were both swollen rod end. Stain FW431 had flagella. FW431 and FW731 were named Clostridium luticellarii and Clostridium cellarii. Followed by Biolog carbon method, the metabolic diversity of isolated strains was analyzed. The results showed that these strains all showed abundant metabolic diversity. According to the results of fermentation, we found strains RCM09 and MRS61 all had wider ability to generate small molecule acid. Strain RCM09 could produce acetic acid, propionic acid, butyric acid and butyl glycol. Strain MRS61 could produce acetic acid and propionic acid.The main flavor substances of the strong aroma style Chinese liquor were ethyl butyrate, ethyl hexanoate, ethyl acetate and lactic acid ethyl ester. These strains produced acids liquor wine quality has an important influence. Those acid substance produced by the bacteria in the cellar mud are important for the liquor’s flavor. Therefore, this study not only enrich microbial resources and promotes the advancement of traditional brewing technology, but also shows important practical significance and theoretical value to further clarify the mud of the cellar bacterial metabolism and fermentation diversity.