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OALib Journal期刊
ISSN: 2333-9721
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产碱杆菌dn25中降氰酶的分离纯化及生化特性
DOI: 10.3724/SP.J.1145.2012.00108, PP. 108-114
Keywords: 产碱杆菌,降氰酶,分离纯化,酶特性,氰化物
Abstract:
以一株可降氰的产碱杆菌dn25为酶来源，通过超滤、30mg/ml硫酸鱼精蛋白沉淀、30%~70%硫酸铵盐析和phenyl-toyopearl650m疏水层析等步骤，获得比活力为44u/mg的纯化酶制剂.在确定酶浓度、反应时间等氰降解活力测定条件后开展酶学性质研究，试图为将来氰降解代谢机理的深入研究和菌株的基因工程改造提供理论基础.研究结果表明，此纯化酶催化氰化物水解的最适ph值为8.0，最适温度为30℃.该酶在ph7.0~8.0区域稳定，而在ph>9时会很快失活；在30℃保存10h，酶活力保持稳定，高于60℃，酶快速失活.加入甘氨酸稳定剂，在60℃下保存20min酶活仍可保留19.6%.酶促反应动力学符合米氏双曲线方程，测得米氏常数km为3.11mmol/l，最大反应速率vmax为0.23mmoll-1min-1.图11表1参23
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