| 腈水解酶重组菌发酵产酶条件的优化及应用 |
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| 引用本文： | 吴乔,薛亚平,郑裕国.腈水解酶重组菌发酵产酶条件的优化及应用[J].化学与生物工程,2012(3):22-26. |
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| 作者姓名： | 吴乔 薛亚平 郑裕国 |
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| 作者单位： | 浙江工业大学生物工程研究所 |
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| 基金项目： | 国家863计划资助项目(2009AA02Z203) |
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| 摘 要： | 优化了重组菌E.coli BL21(DE3)/pET28b(+)-NIT发酵产腈水解酶的条件,确定最适发酵培养基为:玉米浆粉25g.L-1,酵母浸出汁5g.L-1,NaCl 10g.L-1,Fe3+0.5mmol.L-1,初始pH值7.5;最适诱导产酶条件为:诱导剂乳糖0.5g.L-1,诱导剂加入时间为接种6h后,诱导温度30℃,诱导时间28h,在此条件下产酶量达到6161.46U.L-1。所得菌体用于不对称转化R,S-扁桃腈生成R-(-)-扁桃酸,转化率达92.02%,产物e.e.值达99%。
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| 关 键 词： | R-(-)-扁桃酸 重组腈水解酶 乳糖 不对称催化 |
Optimization of Culture Conditions of Nitrilase from Recombinant Escherichia Coli and Its Applicaiton |
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| WU Qiao,XUE Ya-ping,ZHENG Yu-guo.Optimization of Culture Conditions of Nitrilase from Recombinant Escherichia Coli and Its Applicaiton[J].Chemistry & Bioengineering,2012(3):22-26. |
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| Authors: | WU Qiao XUE Ya-ping ZHENG Yu-guo |
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| Affiliation: | (Institute of Bioengineering,Zhejiang University of Technology,Hangzhou 310014,China) |
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| Abstract: | In this paper,culture conditions of recombinant E.coli BL21(DE3)/pET28b(+)-NIT producing nitrilase were optimized.The optimum culture medium was as follows:corn paste powder 25 g·L-1,yeast extract paste 5 g·L-1,NaCl 10 g·L-1,Fe3+ 0.5 mmol·L-1,initial pH value 7.5.0.5 g·L-1 Lactose was added to the culture after inoculating 6 h.The highest nitrilase production reached 6161.46 U·L-1 after inducting for 28 h at 30 ℃.The bioconversion of mandelonitrile by the recombinant nitrilase was also investigated.The result showed that the conversion rate of R-(-)-mandelic acid against substrate reached 92.02% with e.e.value of 99%. |
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| Keywords: | R-(-)-mandelic acid recombinant nitrilase lactose asymmetric catalysis |
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