The Study About The Silage Additives And The Environmental Microorganism Produced Cellulase

Due to the advantages of low cost, high value of nutrition, unrestricted climateand season, and so on, the silage is widely used in the livestock breeding. But theproduction process of silage involves the probiotic action such as metabolism andmaterial degradation, so the probiotic is usually used as an additive of silage toimprove silage fermentation, anticorrosive and enhance its palatability. According toits effect, the silage additives can be divided into three kinds: facilitative additive,inhibitive additive and the nutrient additive. Among of them, the cellulose enzymepreparations often as a nutritional silage additive which can effectively improve thenutritional value of the silage become a hot spot in current study of animal nutrition.In this study, the microbial diversity of the silage in different stages which weretreated with four kinds of silage additives (Numbers for A, B, C, D, institute ofmicrobiology, Chinese academy of sciences) was analyzed respectively by using themethod of16S rRNA clone library, and the effect of four additives in silage process tomaintain stability and oneness silage flora was discussed preliminary. We haveimproved the indirect method to extract DNA, and successfully constructed the16SrDNA clone library, tested the silage samples treated by four different additivesamong1to6months, we analyzed the four samples’ flora diversity in1to6monthand established the phylogenetic tree. The results showed that, after1month, all ofthe samples, the Lactic acid bacteria became the dominant fungi, and after4month,the flora diversity in the four samples is relatively single, and the additive C was thebest one to maintain the flora oneness.During the silage process, the microbial fermentation will cause the temperaturerise and pH drop of silage, so it may be bring certain influence on the additive enzymepreparations. that is why it has been in urgent need of finding the suitable functionalgenetic resources. This study screened a bacteria strain which can produce cellulase from the hot spring water samples in Chenzhou, Hunan Province. This was agram-positive bacteria strain, and according the16S rRNA sequence analysis, it wasbelong to the Paenibacillus sp., and was named CZWQ-32.The research about theculture conditions influenced the enzyme production showed that: the optimumtemperature for cellulase production was50℃, the optimum medium initial pH was5.0, and the optimum carbon resource and nitrogen source were bran and tryptone.The optimum temperature and optimum pH of the cellulase produced by CZWQ-32were60℃and5.0respectively. In addition, the enzyme activity of cellulase wasstable for4hours under the conditions of pH5.0-10.0,60℃, which can be used insilage as the additives.