菌株C3212的系统分类及噬琼胶菌D1326所产琼胶酶研究.pdf

录 摘要 ···························································································· 1 Abstract ························································································ 3 第一章 前言 ················································································· 6 第二章 Leucothrix sargassi C3212 的分类鉴定 ······································ 9 第三章 Agarivorans sp. D1326 的基因组学分析 ···································· 27 第四章 Agarivorans sp. D1326 所产琼胶酶的异源表达及酶学性质 ············ 40 总结 ··························································································· 79 参考文献 ····················································································· 81 综述：琼胶酶的研究进展 ································································ 85 攻读学位期间发表文章情况 ····························································· 96 致谢 ··························································································· 97 个人简历 ····················································································· 98 新乡医学院硕士学位论文 摘 要 背景 海洋生境中蕴含着丰富的具有特殊功能的微生物资源和酶资源。琼胶酶就是一类 主要由海洋微生物产生的糖苷水解酶。由于琼胶酶可水解琼脂糖产生具有特殊生物活 性的琼胶寡糖，使其在食品、医药、化妆品、日用化工和科学研究等领域极具应用潜 力。而现有琼胶酶水解效率低、稳定性差，极大限制了其应用。 目的 为增加琼胶酶种类，丰富现有琼胶酶资源，本论文从海洋环境中分离纯化可培养 细菌，筛选产琼胶酶菌株，采用基因组学和分泌组学相结合的方法最大化挖掘琼胶酶 阳性菌株所产琼胶酶资源。 方法 1. 选用 4 种海洋微生物培养基，分离、纯化黄海近岸海水、海藻样品的可培养 细菌。通过 16S rRNA 基因序列分析确定候选新物种，多相分类学方法确定部分候选 新物种的分类地位。使用卢戈氏碘液染色法及观察琼脂平板降解现象，筛选产琼胶酶 菌株。 2. 采用基因组学和分泌组学相结合的方法挖掘产琼胶酶菌株 D1326 所产琼胶酶， 并利用 DNAMAN 、DNAstar 、SignalP-5.0 Server 等生物信息学软件和服务器对琼胶 酶编码基因序列进行分析，选取代表性的新型琼胶酶作进一步研究。 3. 运用分子生