羟基乙腈水解酶菌株的筛选及其催化特性.PDF

微生物学通报 JUN 20, 2011, 38(6): 825−831 Microbiology China © 2011 by Institute of Microbiology, CAS tongbao@ 研究报告 羟基乙腈水解酶菌株的筛选及其催化特性 * 王利群 陈萦慈 孙晓慧 蔡志强 汪庆 (常州大学制药与生命科学学院 江苏 常州 213164) 摘 要: 以羟基乙腈为唯一氮源, 从土壤中筛选到一株腈水解酶产生菌 CCZU-12, 经形态观察、 生理生化实验和 16S rDNA 序列分析, 鉴定该菌为假单胞菌属(Pseudomonas sp.) 。对菌株CCZU-12 产腈水解酶的培养条件及催化反应条件进行优化, 最适产酶培养条件为: 碳源为 10 g/L 乙酸钠, 氮 源为5 g/L 酵母粉, 金属离子为 1.0 mmol/L Mg2+ , 培养温度30 °C, pH 值7.0, 接种量4%, 装液量 50 mL/250 mL; 最适催化反应温度35 °C, pH 值7.0, 反应 120 h, 羟基乙腈转化率达到98.9% 。 关键词: 羟基乙酸, 腈水解酶, 假单胞菌, 优化 Isolation and catalytic characteristics of a glycolonitrile-hydrolyzing strain * WANG Li-Qun CHEN Ying-Ci SUN Xiao-Hui CAI Zhi-Qiang WANG Qing (School of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, Jiangsu 213164, China) Abstract: A strain CCZU-12 with high glycolonitrile-hydrolyzing activity has been isolated using gly- colonitrile as sole nitrogen source. It was identified as Pseudomonas sp. by its morphological, physio- logical properties and 16S rDNA sequence analysis. The preferred carbon/nitrogen sources and metal ions were sodium acetate (10 g/L), yeast extract (5 g/L), and Mg2+ (1.0 mmol/L), respectively. The op- timum culture conditions were the filling volume 50 mL in 250 mL shaking flask, inoculum 4%, tem- perature 30 °C, and initial pH 7.0. After hydrolysis of glycolonitrile for 120