一株高产壳聚糖酶菌株的筛选和鉴定
首发时间:2017-09-29
摘要:本课题壳聚糖酶这一问题,针对产壳聚糖酶菌株的分离筛选和鉴定进行了研究。首先采集虾蟹壳丰富的海边土壤样品,以胶体壳聚糖为唯一碳源,利用富集培养和透明圈法根据菌株产生的透明圈与菌落直径的比值(D/d)对产壳聚糖酶高产菌株进行初步筛选;最后利用DNS法对所产壳聚糖酶的菌株进行复筛,结合六株菌的持续酶活力测定曲线,最终确定了C4为最佳壳聚糖酶产生菌株,其D/d 值及相应酶活力分别为 4.10±0.22 和 6.235±0.025U/mL。根据形态特征观察菌落形态呈乳白色、圆形、隆起,表面光滑、湿润、边缘平整,周围有透明圈。通过生理生化和分子生物学鉴定,初步鉴定该菌株为Mitsuaria sp.。目前看来,获得高产量、高活性的壳聚糖酶产生菌株为工业化生产提供新酶源的意义重大。
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Screening and identification of a strain with high yield of Chitosan
Abstract:In this study, the isolation, screening and identification of chitosanase-producing strains were studied.Firstly,collecting soil samples from shrimp and crab shell rich, using colloidal chitosan as the only carbon source, by enrichment culture and transparent circle method according to the ratio of strains produce transparent circle and colony diameter (D/d) were used for preliminary screening of strains;secondly, screening strains by DNS method combined with enzyme activity, and ultimately determining the C4 strain is the best, the D/d value and the corresponding enzyme activity is 4.10 + 0.22 and 6.235 + 0.025U/mL.According to the morphological characteristics, the morphology of the colonies was milky white, round and raised, the surface was smooth and moist, the edge was smooth, and there was a transparent circle around it.With studying phenotypical, genotypical and phylogenetical, based on sequence analysis of 16SrDNA genes, and finally identitied the strain as Mitsuaria sp.so far,providing a new source of high yield chitosanase-producing strains for industrial production is great significance. ?????
Keywords: chitosan sification and identification Mitsuaria sp.
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