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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

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曹明乐 1   姜兴林 2   张海波 2 *   咸漠 2   徐鑫 2   刘炜 2  
文摘 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.
其他语种文摘 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.
来源 应用与环境生物学报 ,2013,19(2):346-350 【核心库】
DOI 10.3724/sp.j.1145.2013.00346
关键词 腈水解酶 ; 泛菌 ; 生物催化 ; 生物治理
地址

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

语种 中文
文献类型 研究性论文
ISSN 1006-687X
学科 生物化学;微生物学
基金 山东省自然科学基金 ;  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
文献收藏号 CSCD:4835908

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引证文献 2

1 龚劲松 腈水解酶在医药中间体生物催化研究中的最新进展 化学进展,2015,27(4):448-458
被引 4

2 王世伟 原核微生物腈转化酶研究进展 中国生物工程杂志,2015,35(10):100-107
被引 1

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