Site-saturation mutagenesis of formate dehydrogenase from Candida bodinii creating effective NADP(+)-dependent FDH enzymes
文献类型：期刊论文
| 作者 | Wu, Weihua2; Zhu, Dunming1,2; Hua, Ling2 |
| 刊名 | JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
 |
| 出版日期 | 2009-12-01 |
| 卷号 | 61期号:3-4页码:157-161 |
| 关键词 | Candida bodinii Cofactor specificity Formate dehydrogenase NADP(+)-dependent formate dehydrogenase Site-saturation mutagenesis |
| 英文摘要 | The analysis of previous reported results envisioned that residues Asp195, Tyr196 and Gln197 of formate dehydrogenase from Candida bodinii (CboFDH) might play critical roles in determining the enzyme's cofactor specificity. With the aim to develop novel NADP(+)-dependent formate dehydrogenase enzymes, simultaneous site-saturation mutagenesis of residues Asp195 and Tyr196 of CboFDH coupled with screening resulted in two mutant enzymes, D195Q/Y196R and D195S/Y196P, which showed significant NADP(+) specificity. The overall catalytic efficiencies (k(cat)/K-m) toward NADP(+) were 1.14 x 10(4) and 2.9 x 10(3) M-1 s(-1), respectively, which are higher than the reported mutant CboFDHs obtained by sequential mtltagenesis. The ratio of catalytic efficiencies (k(cat)/K-m)(NADP+)/(k(cat)/K-m)(NAD+) of D195Q/Y196R and D195S/Y196P were 2.1 and 0.2, respectively. Mutation of residue Gln197 of D195Q/Y196R to Asn further increased the enzyme's overall catalytic efficiencies (k(cat)/K-m) toward NADP(+) to 29.1 x 10(3) M-1 s(-1), with (k(cat)/K-m)(NADP+)/(k(cat)/K-m)(NAD+) being 17.1, which are much higher than the reported data for a mutant enzyme of formate dehydrogenase from Pseudomanas sp. 101 (PSeFDH). This study demonstrates that residues 195. 196 and 197 really play critical roles in determining the enzyme's cofactor specificity. (C) 2009 Elsevier B.V. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences |
| 类目[WOS] | Biochemistry & Molecular Biology ; Chemistry, Physical |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Chemistry |
| 关键词[WOS] | HIGH-RESOLUTION STRUCTURES ; NADP-DEPENDENT ENZYMES ; COENZYME SPECIFICITY ; CARBONYL REDUCTASE ; BINDING-SITE ; BOIDINII ; ENANTIOSELECTIVITY ; METHYLICA ; KETONES ; NAD(+) |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000272064600007 |
| 源URL | [http://124.16.173.210/handle/834782/1463]  |
| 专题 | 天津工业生物技术研究所_生物催化与绿色化工 朱敦明_期刊论文 |
| 作者单位 | 1.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, State Engn Lab Ind Enzymes, Tianjin 300308, Peoples R China 2.So Methodist Univ, Dept Chem, Dallas, TX 75275 USA |
| 推荐引用方式 GB/T 7714 | Wu, Weihua,Zhu, Dunming,Hua, Ling. Site-saturation mutagenesis of formate dehydrogenase from Candida bodinii creating effective NADP(+)-dependent FDH enzymes[J]. JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC,2009,61(3-4):157-161. |
| APA | Wu, Weihua,Zhu, Dunming,&Hua, Ling.(2009).Site-saturation mutagenesis of formate dehydrogenase from Candida bodinii creating effective NADP(+)-dependent FDH enzymes.JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC,61(3-4),157-161. |
| MLA | Wu, Weihua,et al."Site-saturation mutagenesis of formate dehydrogenase from Candida bodinii creating effective NADP(+)-dependent FDH enzymes".JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC 61.3-4(2009):157-161. |
入库方式： OAI收割
来源：天津工业生物技术研究所
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