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Tempeh originated in China, is one of China's ancient fermented soybean products, which is based on black beans or soybeans as raw material, the use of microbial fermentation of a traditional non-staple food seasoning, soy fragrant attractive, unique flavor and are popular among consumers parties are welcome. Tempeh has a high nutritional value, rich in soluble sugar, soluble nitrogen, vitamin A, vitamin E, vitamin B group-containing nutrients; also contains soy isoflavones, soy oligosaccharides, soybean saponin, soy peptides, tempeh fibrinolytic enzymes and a variety functional factors. Tempeh not only nutritional value, but also has of detoxification, Chufan Xuan Yu efficacy of treatment seasons fever, headache, chills and fever, chest boredom, imaginary trouble, insomnia, loss of appetite, indigestion, etc.. Tempeh can also prevent Alzheimer's disease, osteoporosis, high blood pressure, diabetes and other age frequently-occurring disease. The Fibrinolytic Enzyme is a hot spot for domestic research, because it is a new type plasminogen can be found in the tempeh directly fibrinolytic, and has a high fibrinolytic activity, the use of safe, reliable, so at home and abroad the attention of scholars. Domestic tempeh Sichuan Yongchuan tempeh, Hunan Liuyang tempeh, the Shandong Linyi tempeh famous. About few Yunnan tempeh, Yunnan is China's largest minority provinces, each nation has retained the traditional process of fermented foods, tempeh more types. In this study, the collection of strains of the four cities of Yunnan Province (Kunming, Yuxi, Zhaotong, Yuanyang) tempeh, tempeh microbial species for a preliminary study, preliminary analysis by SDS-Zymography experiments, tempeh contains protease, and fibrinolytic activity in the tempeh to make a preliminary determination. This study was designed by a preliminary study on the microbial diversity of different selective solid medium and biochemical experiments tempeh, a preliminary identification of the the tempeh samples of microbial. Establish an effective DNA extraction system, the application 16SrDNA universal primers for PCR amplification, sequence alignment sample microbial species identification and the establishment of a phylogenetic tree, to explore and analyze the microbial species diversity in Yunnan Province, tempeh . Tempeh protease activity in the preliminary study, the application of protease activity assay and SDS-PAGE-were speculating that the difference in crude experiments, analysis proteases in the sample and the level of activity, summed up the different parts of tempeh the type and activity of the protease. Different regions of Yunnan Province, sample collection and experimental summary of the tempeh Microbial Diversity in different regions to discuss the protease activity and types in the sample, which found useful microbial resources. Applications such as SDS-Zymography experimental protease tempeh contains preliminary analysis, based on the number of plastic sheet on the strip and brightness can be initially identified the type of protease in the sample, the results of Kunming strain, higher protease activity, secretion The protease has the largest number of molecules, approximately nine species of varying molecular weight protease, a molecular weight of about 14KDa-116KDa uniformly distributed, the inference may contain a variety of proteases, such as gelatinase, collagenase, tempeh plasminogen; Yuanyang 11 (1) No. strains Kunming 9 similar, the largest number of molecules secreted protease, about 11 species of varying molecular weight protease; of Yuxi 1,3 and No. 9 6 strains secreted protease largest number of molecules, about 5 molecular weight of not and other proteases; Zhaotong 25 strains protease secreted by the number of molecules up to about three kinds of varying molecular weight protease. In summary, the analysis contained in the four regions of strain type and activity of protease, especially Kunming strain 9 and $ Yang strain 11 (1) the most prominent, two strains on the secretion of protease molecules, and protease activity were the highest. Application of improved agar - fibrin plate experiment, the calculated size of the soluble protein circle, and according to the experimental results of SDS-fibrin-PAGE and ninhydrin determination of protease activity is obtained, wherein the Yuxi YX-1 strain protease activity 96.22U/mL YX-11 strain protease activity of 93.88U/mL YX-18 strain protease activity of 96.66U/mL Zhaotong ZT-13 strain protease activity of 97.09U/mL, ZT-15 strain protease activity 94.61U/mL , which speculate the Fibrinolytic Enzyme activity Yuxi tempeh sample is the highest among the four regions, followed Zhaotong. The experimental results for future in-depth study has laid a solid foundation for high plasmin activity tempeh samples.
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