细菌型豆豉后发酵期间菌相研究及产蛋白酶菌种的筛选-study on bacterial phase and screening of protease-producing strains during post-fermentation of bacterial douchi.docxVIP

细菌型豆豉后发酵期间菌相分析及蛋白酶菌种的筛选 细菌型豆豉后发酵期间菌相分析及蛋白酶菌种的筛选 4 4 The composition of the bacteria during the post-fermentation of Douchi were analyzed by using microscopy and colony morphology. The results showed that, the different types of bacteria occured in different periods, and the proportion of each specie had significantly changed at different times. Meanwhile, forty five different bacteria were isolated and analyzed by restriction fragment length polymorphism analysis (ARDRA). The six types were checked out from those forty five strains, the 16S rDNA sequences of representative strains of the various types were amplified and aligned. The results showed that, the proportion of Bacillus subtilis was 89%, and Bacillus amyloliquefaciens was 4%. There were Micrococcus luteus and Staphylococcus gallinarum during the post-fermentation of Douchi. According to the enzyme activity and the content of free nitrogen of starter of Lobster Sauce, five high-yield protease-producing strains were screened by casein plate and re-screened by starter-making from forty five strains. The five superior strains were used to ferment Lobster Sauce respectively. According to the sensory tests, physical and chemical indexes of five products, the Lobster Sauce fermented by D2 strain had a good flavor and the product was very delicious. By physiological characterization and the sequence analysis of 16S ribosomal DNA genes, the strain D2 was preliminarily identified as Bacillus subtilis. The strain was got from Lobster Sauce and it was safe, so it had further studying and developing value. Key words: Bacterial Douchi, DGGE, T-RFLP, Bacterial flora, High-yield protease-producing bacterial strains 关于学位论文原创性和使用授权的声明 本人所呈交的学位论文，是在导师指导下，独立进行科学研究所 取得的成果。对在论文研究期间给予指导、帮助和做出重要贡献的个 人或集体，均在文中明确说明。本声明的法律责任由本人承担。 本人完全了解山东农业大学有关保留和使用学位论文的规定，同 意学校保留和按要求向国家有关部门或机构送交论文纸质本和电子 版，允许论文被查阅和借阅。本人授权山东农业大学可以将本学位论 文的全部或部分内容编入有关数据库进行检索，可以采用影印、缩印 或其他复制手段保存论文和汇编本学位论文。 保密论文在解密后应遵守此规定。 论文作者签名： 导 师 签 名： 日 期： 山东农业大学硕士学位论文 山东农业大学