flavobacterium johnsoniae 昆布多糖酶的分离纯化及其催化性质.pdf

微生物学通报 JUN 20, 2011, 38(6): 839−846 Microbiology China © 2011 by Institute of Microbiology, CAS tongbao@ 研究报告 Flavobacterium johnsoniae 昆布多糖酶的分离 纯化及其催化性质 周亮 朱永涛 陈冠军 刘巍峰* (山东大学 微生物技术国家重点实验室 山东 济南 250100) 摘 要: 约氏黄杆菌Flavobacterium johnsoniae 具有分泌裂解酵母细胞壁酶系的能力, 经初步分析 发现其发酵液中具有葡聚糖酶、几丁质酶和蛋白酶等活性。通过离子交换层析、疏水层析和凝胶 过滤层析, 从该菌发酵液中分离纯化到一种昆布多糖酶。该酶分子量为 35 kD 左右, 其最适反应 温度为50 °C, 最适反应pH 为5.0。以昆布多糖和昆布寡糖为底物的反应表明, 该酶以内切酶作用 模式进行催化水解。 关键词: 约氏黄杆菌, 葡聚糖, 酵母细胞壁, 昆布多糖酶, 纯化 Purification and catalytic properties of a laminarinase from Flavobacterium johnsoniae * ZHOU Liang ZHU Yong-Tao CHEN Guan-Jun LIU Wei-Feng (The State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong 250100, China) Abstract: Flavobacterium johnsoniae is capable of secreting enzymes which can efficiently hydrolyze yeast cell wall. Preliminary analysis revealed the presence of glucanase, chintinase and protease activi- ties in its culture supernatant. A laminarinase was purified from the extracellular components of F. johnsoniae through several isolation steps including ion exchange, hydrophobic interaction and gel ex- clusion chromatography. The molecular weight of the purified laminarinase is about 35 kD. The opti- mum temperature and pH of its catalyzed hydrolysis