一株产腈水解酶的泛菌及其催化特征  

A New Nitrilase Producing Pantoea sp. and Its Enzymological Properties

作　　者：曹明乐[1] 姜兴林[2] 张海波[2] 咸漠[2] 徐鑫[2] 刘炜[2] 

机构地区：[1]山东大学微生物技术国家重点实验室,济南250100 [2]中国科学院青岛生物能源与过程研究所,青岛266101

出　　处：《应用与环境生物学报》2013年第2期346-350,共5页Chinese Journal of Applied and Environmental Biology

基　　金：山东省自然科学基金项目(Y2008B43);中国科学院知识创新工程工业生物技术领域基础前沿研究专项(Y112131105)资助~~

摘　　要：通过初步活化和分步胁迫富集,采用Berthelot法高通量筛选与高压液相精细筛选获得一株底物广泛性良好的产腈水解酶菌株Pantoea sp.;通过菌株降解乙腈(脂肪腈类)、3-羟基丙腈(羟基腈类)、4-羟基苯乙腈(芳香腈类)等不同种类腈类,表明该菌株具有较好的底物广泛性;通过单因素法优化菌株培养条件,最佳碳源为壳聚糖,最佳氮源为无机氮源硫酸铵,产酶量达到16.45 U/mL.向反应体系中加入终浓度为1 mmol/L的钴、锌、钙离子可以增强酶活.酶学性质研究表明该腈水解酶的最适反应pH为7.0,其中在pH 6.6-7.6之间该菌株酶活性能够保持在最高酶活性的80%以上;最适反应温度为30℃,在22-45℃范围内催化活性大于最大酶活性的80%,反应30 h后离心菌体二次催化几乎不丧失活性.该菌株在医药、食品、农业、环境污染治理等领域具有潜在的工业应用开发前景.Nitrilases can hydrolyze nitrile efficiently under mild conditions. For their low pollution and cost, the enzymatic methods with nitrilases are widely used in agriculture, industry, biomedicine and environment protection. Nitrilases with broad substrate specificity are lacked in the nitrilase market. In this study, Berthelot method and high performance liquid chromatography were used to separate and identify a new strain of nitrilase Pantoea sp. 1-2, which can hydrolyze acetonitrile, 4-hydroxyphenylcyanide and 3-hydroxypropionitrile. The strain produced 16.45 U/mL nitrilase with ammonium sulfate as nitrogen source. The enzyme remained 80% activity at pH 6.6-7.6, temperature 22-45 ℃, staying stable after 30 h incubation. The strain would be of great value in industrial utilization. Fig 7, Ref 21

关 键 词：腈水解酶 泛菌 生物催化 生物治理 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]