| 易错PCR法定向进化D-海因酶的初步研究 |
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| 引用本文： | 孔荣,钮利喜,袁静明.易错PCR法定向进化D-海因酶的初步研究[J].山西大学学报(自然科学版),2006,29(4):425-427. |
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| 作者姓名： | 孔荣 钮利喜 袁静明 |
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| 作者单位： | 山西大学,生物技术研究所,山西,太原,030006 |
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| 摘 要： | D-海因酶是生物转化D-型氨基酸的关键酶,在一定浓度M n2 存在下,经易错PCR法重复扩增,产物构建成pET-HDT重组子,并建立突变体文库.经筛选获得了内源DNA序列变异的变异株.进化酶催化底物海因水解的活性为亲本重组酶的1.3倍,催化底物对羟基苯海因水解的活性为亲本重组酶的2.4倍.
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| 关 键 词： | 海因酶 易错PCR 酶活性 |
| 文章编号： | 0253-2395(2006)04-0425-03 |
| 收稿时间： | 2005-09-12 |
| 修稿时间： | 2005-12-06 |
A Preliminary Study on the Directed Evolution of D-hydantoinase by Error-prone PCR |
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| KONG Rong,NIU Li-xi,YUAN Jing-ming.A Preliminary Study on the Directed Evolution of D-hydantoinase by Error-prone PCR[J].Journal of Shanxi University (Natural Science Edition),2006,29(4):425-427. |
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| Authors: | KONG Rong NIU Li-xi YUAN Jing-ming |
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| Institution: | Institute of Biotechnology , Shanxi University, Taiyuan 030006, China |
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| Abstract: | D-hydantoinase(HDT) plays an important role in the bioconversion of D-amino acids.The enzyme gene was repeatedly amplified by error-prone PCR at a definite concentration of Mn~(2 ) and PCR products were cloned to vector pET-3a to form the recombinant plasmid pET-HDT,followed by generating a mutant molecular library.Positive colony was selected to confirm the gene mutation by DNA sequence analysis.The mutant enzyme showed its activity as 1.3 folds as the original recombinant enzyme with DL-hydantoin as the substrate,whilst as 2.4 times with D-p-hydroxyophenylhydantoin as the substrate. |
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| Keywords: | Hydantoinase error-prone PCR enzymatic activity |
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