| 一种3-氰基吡啶水解酶产生菌的高通量筛选策略及应用 |
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| 引用本文： | 周琼,林春昕,蔡志强,何玉财,赵希岳,鲍磊,鲍春芬.一种3-氰基吡啶水解酶产生菌的高通量筛选策略及应用[J].生物加工过程,2011,9(6):30-34. |
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| 作者姓名： | 周琼 林春昕 蔡志强 何玉财 赵希岳 鲍磊 鲍春芬 |
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| 作者单位： | 常州大学制药与生命科学学院生物工程研究室,常州,213164 |
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| 基金项目： | 国家自然科学基金资助项目，常州市生物医药科技专项基金资助项目，常州大学人才引进基金资助项目，常州大学大学生创新基金资助项目 |
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| 摘 要： | 利用改进的羟肟酸铁分光光度比色法建立了一种简单、快速、高通量的腈水解酶筛选方法.应用该方法从土壤中筛选获得1株具有3-氰基吡啶水解酶活性的菌株CCZU10 -1,经16S rDNA序列分析,鉴定该菌为红球菌属Rhodococcus sp.；同时确定了最适反应温度、pH和金属离子添加剂分别为30℃、7.0和Ca2+ (0.1 mmol/L).在最适催化反应条件下,催化转化50 mmol/L烟腈36 h,烟酸的产率可达到93.5％.
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| 关 键 词： | 3-氰基吡啶 羟肟酸铁 烟酸 腈水解酶 高通量筛选 |
High-throughput screening for 3-cyanopyridine-hydrolyzing enzyme producing strain and its application |
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| ZHOU Qiong,LIN Chunxin,CAI Zhiqiang,HE Yucai,ZHAO Xiyue,BAO Lei,BAO Chunfen.High-throughput screening for 3-cyanopyridine-hydrolyzing enzyme producing strain and its application[J].Chinese Journal of Bioprocess Engineering,2011,9(6):30-34. |
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| Authors: | ZHOU Qiong LIN Chunxin CAI Zhiqiang HE Yucai ZHAO Xiyue BAO Lei BAO Chunfen |
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| Institution: | ZHOU Qiong,LIN Chunxin,CAI Zhiqiang,HE Yucai,ZHAO Xiyue,BAO Lei,BAO Chunfen(Laboratory of Biochemical Engineering,College of Pharmaceutical and Life Sciences,Changzhou University,Changzhou 213164,China) |
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| Abstract: | A high-throughput screening method for nitrile-hydrolyzing enzyme was used based on a modified ferric hydroxamate spectrophotometry.Using 3-cyanopyridine as a sole nitrogen source,a nitrile-hydrolyzing strain was isolated from soil.It was identified as Rhodococcus sp.by 16S rDNA sequence analysis.Moreover,the optimum reaction temperature,pH and metal ion additive were 30 ℃,7.0 and Ca2＋（0.1 mmol/L）,respectively.After 36 h of biotransformation,the yield of nicotinic acid from the hydrolysis of 3-cyanopyridine（50 mmol/L） reached 93.5%. |
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| Keywords: | 3-cyanopyridine ferric hydroxamate nicotinic acid nitrile-hydrolyzing enzyme high-throughput screening |
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