（毕业设计论文）《产壳聚糖酶海洋微生物菌株的筛选、鉴定与产酶条件研究》.doc

目录 中文摘要 I Abstract II 引言 1 1 实验材料和方法 3 1.1 主要仪器与设备 3 1.2 实验药品和试剂 3 1.3 实验方法 4 1.4 实验步骤 5 2 结果与分析 9 2.1 富集培养与平板筛选 9 2.2 菌株KJT-A鉴定和分析 9 2.3 菌株KJT-D鉴定与分析 12 2.4 菌株KJT-A的生长情况 14 2.5 菌株KJT-D的生长情况 15 2.6 假单胞菌株KJT-A壳聚糖酶的分离 16 2.7 GF1-4的SDS检测 17 3 小结与讨论 19 3.1 产壳聚糖酶海洋微生物菌株的筛选 19 3.2 产壳聚糖酶菌株的鉴定 19 3.3 温度与盐度对产壳聚糖酶菌株生长的影响 20 3.4 壳聚糖粗酶的制备和纯化 20 4 结论 22 参考文献： 23 附录Ⅰ 26 附录Ⅱ 28 附译文 30 附译文原文 34 致 谢 38 产壳聚糖酶海洋微生物菌株的筛选、鉴定与产酶条件研究 [摘要] 采用不同培养基对海水养殖环境样品进行壳聚糖酶产生菌的分离，结果表明海水培养基对于菌株筛选有显著的促进作用，同时甲壳类外壳是壳聚糖酶产生菌的良好来源。KJT-A和KJT-D是从海水养殖场蟹壳表面分离到的两株壳聚糖酶产生菌株，结合形态学观察、生理生化反应和分子生物学鉴定，初步确定菌株KJT-A和KJT-D分别为假单胞菌属（Pseudomonas）和青霉属（Penicillium）分析不同温度、盐度对两株菌株生长的影响[关键词] 壳聚糖酶；鉴定；16S rDNA；假单胞菌；青霉 Screening and Identification of Chitosan-Hydrolytic Microorganisms From Marine Environment and Study on the Optimal Conditions of Chitosanase Formation [Abstract] In this paper the chitosanase-producing microbes from the marine environment were studied. The result showed that sea water medium can significantly improve isolation of strain,and the surface of crab shell was best source of chitonsanase-producing bacterial strain. KJT-A and KJT-D, which were isolated from the crab shell surface of xuwang mariculture zone in zhoushan island, were classified into Pseudomonas and Penicillium based on morphology, physiology, biochemistry and molecular biology methods.Exploring the growth of two strains in different temperature and salinity ,we found that KJT-A is a moderately halophilic bacteria,the optimum temperature is 19℃,achieving the maximum absorbance when cultured 48h.KJT-D is a widesalt-resistant strains,the optimum temperature is 28℃,achieving the maximum absorbance at 72h.The two strains can grow well in the normal salinity condition,this is helpful to the sea strains fermentation,and have a great effect to save the freshwater resources.Chitosanase of the above two strains were isolated and purified,which help us have a better understanding of its molecular structure and salinity adaptation,and provide a theoreti