Screening And Application Of Pyrazine-based Bacillus Sp

Moutai-flavor liquor get the majority of consumers because of its outstanding sauce, fragrant and tire, elegant and delicate, mellow wine, soft cotton rich, full flavor long, empty cup fragrant lasting features. Cui Sha Moutai-flavorliquor belonging to Moutai-flavor liquor, compared with the traditional Moutai-flavor liquor, the production of Cui Sha Moutai-flavor liquor had the following advantages: short fermentation time, high liquor yield, short storage period, rapid fund turnover, and cheap product price and high liquor quality. However, its quality was still inferior to Moutai-flavor liquor. This thesis is mainly of isolated and purified the bacteria from Jiang Xiang high-temperature Daqu, screening of high yield of pyrazines function Bacillus by simulating the fermentation process of Moutai-flavor liquor production,and then applicating the functional bacillus to the Cui Sha Moutai-flavor r liquor production. The main results are as follows:1)By the methods of Dilution plate coating from Jiang Xiang Daqu,19 bacte rial strains were isolateed and screening. Combined the morphology, physiology of strain with biochemical and molecular biology, and comprehensive analysis of the situation,FBKL1.0185, FBKL1.0187, FBKL1.0188, FBKL1.0194, FBKL1.0195,FBKL1.0197,FBKL1.0198, FBKL1.0202 belongs to the Bacillus amyloliquefaciens,FBKL1.0196,FBKL1.0203 belongs to Bacillus subtilis, FBKL1.0189, FBKL1.0190 belongs to Bacillus methylotrophic, belongs to FBKL1.0186, FBKL1.0191,FBKL1.0192, FBKL1.0193, FBKL1.0199, FBKL1.0200, FBKL1.0201 belongs to Bacillus licheniformis.2)By simulating the fermentation process of Maotai flavor liquor production,and taking sensory evaluation and GC-MS analysis of their fermentation products, we found that producing relatively rich sauce flavor strain their material content pyrazinees were higher especially tetramethyl pyazine. Wherein the two strains FBKL1.0199 and FBKL1.0201 fermentation products have the highest content of the pyrazine subst-ances, so choose FBKL1.0199 and FBKL1.0201 as functional strains which has the ability of high-yielding pyrazine substances. By using different proportion combina-VIII tion of The FBKL1.0199 and FBKL1.0201 to simulated the solid state fermentation experiment, the results showed that FBKL1.0199 and FBKL1.0201 ratio was 1:1, the effection was the best.3)Using solid phase microextraction(HS-SPME)combined with GC-MS for the qualitative analysis of tetramethyl pyrazine which was found in Maotai-flavor liquor and fermenting grains. And determined the tetramethylpyazine peak time in the chromatographic conditions. By headspace solid phase micro extraction(HS- SPME)optimized conditions, obtained tetramethylpyazine optimal extraction conditions: extraction temperature 60?, extraction time 40 min, Na Cl dosage 1.5g. Quantitative method for bran Maotai-flavor liquor and fermenting grains of tetramethyl pyrazine was established. The relative standard deviation of the method was 7.74%, the recovery was between 89.2% ~ 106.4%, met the requirements of the analysis of the sample, this method high reliability, it can be used to analyze and determine the content of tetramethylpyrazine in fermented grains and wine samples.4)The functional strains FBKL1.0199 and FBKL1.0201 was add to the Cui Sha Maotai-flavor liquor production. Fermentation conditions were optimized by single factor experiment and orthogonal experiment, to study when the fermentation finished,the tetramethylpyazine content of Cui Sha Maotai-flavor liquor and fermenting grains and wine-like taste. The results showed that the optimum fermentation conditions of FBKL1.0199 and FBKL1.0201 was that inoculation ratio is 5%, the grains ratio is 1:5,and the fermented p H value of 6.0.The distilled from wine samples in tetramethyl pyrazine content was the highest, reaching 6.34mg/L.By compared with blank group which was not add the functional Bacillus, the tetramethyl pyrazine content of experimental group which was added the functional Bacillus and optimizated the fermentation conditions, was significantly increased and the wine quality clear transparent, wine body coordination, less bitter, Maotai flavor prominent, the feeling is mach better.5)Through the determination of protein content in the fermented grains before and after fermentation, we found that after fermentation, protein content in the fermented grains were lower than before the fermentation.And through add thefunctional Bacillus in the experimental group, after the fermentation fermented grains protein content was lower than which did not join the functional Bacillus to fermented.The reasons maybe as follows: First, the most proteins are used by microorganism to maintain its growth and metabolism. Second, a few of proteins was transformed to another substances and some pyrazine through the metabolic activities of functional bacteria.