一株海洋细菌Renibacterium+sp.QD1中的壳聚糖酶及产酶条件的研究.pdf

到的纯壳聚糖酶进行了蛋白质N 端测序，获得了Csn-A 的N 端 15 个氨基酸的序 列。对该段氨基酸序列进行比对和分析发现，其与Renibacterium salmoninarum （鲑鱼肾杆菌）中的一个壳聚糖酶具有很高的同源性，而且 16srDNA 序列也显 示Renibacterium sp. QD1 与Renibacterium salmoninarum 的种属同源性达99% 以上， 推测两株菌的产酶序列上较为接近。因此我们利用 Renibacterium salmoninarum 已知的产壳聚糖酶基因序列设计引物进行PCR 试验，结果得到了 一段 944 碱基的序列，与 Renibacterium salmoninarum 产酶基因序列相似度达 95%，与已经报道的Streptomyces N174 产酶基因序列相似度达67%，并且该序 列为糖苷水解酶46 家族。对csn-A 基因的克隆与分析，无疑丰富了壳聚糖酶基 因的资源，推动了壳聚糖酶构效关系的研究。 关键词：壳聚糖酶；分离纯化；酶学性质；基因克隆；发酵优化 2 Screening, identification and research on enzyme production of the chitosanase produced from marine bacteria Renibacterium sp. QD1 Abstract Chitosan, an important marine polysaccharide, is limited to be used due to its low solubility, high molecular weight, and absorption rate. Chitooligosaccharides can be prepared through chemical method and physical method. However, the hydrolysis product is not homogenous and the hydrolyzing reaction is uncontrollable. Those methods also causing serious environmental pollution. Enzymatic degradation of chitosan should be a promising alternative to chemical method and physical method with high specificity and under mild conditions. Especially for it’s low cost and no pollution, It is significante to do research of chitosanase . Currently, it was found a wide range of chitosanase. But because most of the chitosanase activity is low, and poor degradation efficiency, it is difficult to be applied to the industry for the preparation of chitooligosaccharides. Therefore, finding and screening yield novel and high activity chitosanase strains become a hot topic in the field of chitosan research.. Renibacterium sp. QD1, a bacteria strain capable of hydrolyzing chitosan