对羟基苯乙腈水解酶产生菌的筛选及产酶条件研究  

Screening for p-hydroxyphenylcyanide nitrilase and study of enzyme production conditions

作　　者：吴明火[1] 蔡谦[1] 郑裕国[1] 沈寅初[1] 

机构地区：[1]浙江工业大学生物工程研究所,杭州310014

出　　处：《生物加工过程》2005年第4期32-35,44,共5页Chinese Journal of Bioprocess Engineering

基　　金：国家重点基础研究发展规划项目国家"973"项目(No.2003CB716005)

摘　　要：以对羟基苯乙腈为唯一氮源,从土壤中筛选到5株腈水解酶产生菌.在初筛的过程中,用薄板层析(TLC)对细胞转化液定性检测,高效液相色谱法(HPLC) 定量分析.考察了培养时间、不同碳源对细胞酶产量的影响及在反应过程中金属离子对细胞酶产量的影响,实验结果发现,培养时间在70 h左右时细胞酶产量最高,以葡萄糖为碳源质量,质量浓度在12.5～15 g/L附近酶产量最高.Cu2+对细胞酶活力具有非常强烈的抑制作用 ,Co2+、Hg2+、Ni+等对细胞酶活力具有明显的抑制,而Ba2+、Mg2+具有较明显的促进作用,但在高浓度下促进作用减弱.Five strains with high activity of nitrilase to hydrolyzing p-hydroxybenzylcyanide have been isolated through screening and selection. The effect of culture time, carbon source, metal ions on the yield of enzyme were studied. When the culture time near 70 h, the glucose as carbon source with the concentration around 12.5～15 g/L, the yield of enzyme was highest. The Cu^2＋ ion inhibited the enzyme activity strongly and the inhibition of Co^2＋, Hg^2＋, Ni^2＋ was also obvious. But Ba^2＋ and Mg^2＋ ions could accelerate the hydrolysis of p-hydroxybenzylcyanide to p-hydroxyphenylacetic acid.

关 键 词：腈水解酶 腈水合酶 对羟基苯乙腈 对羟基苯乙酸 

分 类 号：Q814[生物学—生物工程]