Isolation And Identification Of Actinomycetes In The Fermentation Bed And Optimization Of Fermentation Condition Of 8F3

With the development of breeding industry, the problem of animal fecal pollution becomes more and more serious. But the traditional treatments of drove’s feces cost too much manpower and material resources, we urgently need to find a new culture model with the characteristic of less input and high output. Fermentation bed is a new culture model which emerged in recent years, has zero discharge, zero pollution, less energy consumption. It becomes more and more important in people’s graces by its advantages which can save energy, improve the quality of meat, and improve economic benefit. In fact, fermentation bed culture model is a degradation process of microorganisms. So the study of microorganisms in the fermentation bed can help us find the essence of the fermentation bed and manufacture new biological agents. In this research, we used the method which combined routine isolation and culture method with modern molecular biology technology PCR-DGGE to isolate and identify the dominant actinomycetes of the fermentation bed. We obtained the following results:（1） By DNA extraction, PCR-DGGE, gene clone and sequencing, we got 11 bands, including 2 dominant bands, 3 secondary dominant bands. And we got 9 sequencing results by BLAST in NCBI, they were Mycobacterium fallax （ AF480600 ） , Corynebacterium xerosis （AM233487） , Actinocorallia aurantiaca （AJ293701）、Janibacter sp. DE08 （ DQ227675 ） known strains, 4 strains were uncultured microorganisms, 1 strain was micrococcus.（2） We obtained 72 strains of medium-temperature actinomycetes. And identified 20 typical strains in colony characteristics, micromorphology characteristics, physiological and biochemical characteristics.（3） In this research we studyed the method of inhibit bacteria contamination, and the result indicated that: sterilized 1h at 120℃, meanwhile added 100mg/L K₂Cr₂O₇ in the medium can effectively prevent the pollution of bacteria and fungi.（4） We obtained 4 strains which produced proteinase, and one of them had stronger ability of producing proteinase; 1 strain produced tyrosinase; 4 strains can decompose cellulose.（5） We got one strain which had stronger ability of producing proteinase, we named it 8F3. The results showed that the most favorable conditions for 8F3 to product proteinase were initial pH value in medium was 7.0, 40mL medium in 250mL flask, and seed inoculation rate was 3%, fermentation for 72 hours at 37℃.