Cavity engineering-driven switching of cofactor specificity in a carbonyl reductase for the synthesis of chiral aryl β-Hydroxy α-amino acid
文献类型：期刊论文
| 作者 | Tang, Jiawei1; Zhong, Jirui2,4; Chen, Hao1; Zhang, Luwen1; Shao, Junxi3; Chen, Haodong1; Zhang, Fuli1; Li, Hongchun5; Chen, Shaoxin1 |
| 刊名 | BIOORGANIC CHEMISTRY
 |
| 出版日期 | 2025-12-01 |
| 卷号 | 167页码:9 |
| 关键词 | Carbonyl reductases Protein engineering Cofactor specificity reversal Chiral aryl beta-hydroxy alpha-amino acids Biocatalysis |
| ISSN号 | 0045-2068 |
| DOI | 10.1016/j.bioorg.2025.109295 |
| 通讯作者 | Li, Hongchun(lihongchun@zidd.ac.cn) ; Chen, Shaoxin(sxzlb@263.net) |
| 英文摘要 | Carbonyl reductases (CRs) are important enzymes with a broad spectrum of industrial applications. However, the CRs reported for use in the asymmetric synthesis of chiral aryl beta-hydroxy alpha-amino acids are exclusively specific to NADPH, posing challenges for cost-effective NADH regeneration. In this study, we focused on the carbonyl reductase (M30) from Exiguobacterium sp. F42 and successfully altered its cofactor preference to favor the more economical NADH using a computer-aided design approach. Through protein engineering, we developed an enhanced mutant M36, S10A/Y15R/E16A/K19L/A32D/R33I, which increased the cofactor specificity of M30 by over 1000-fold. This best combinatorial mutant enabled the highly efficient synthesis of the chloramphenicol intermediate (2S,3R)-5b, achieving 99 % conversion from 50 g/L of substrate 5a with excellent stereoselectivity (ee > 99.9 %, de > 99.9 %), hereby significantly enhancing its industrial application potential. Molecular dynamics (MD) simulations provided atomic-level insights into the mechanistic basis for the altered cofactor affinity and revealed the individual contribution of key mutations. This work establishes an efficient approach for engineering cofactor specificity in CRs and underscores the potential of cavity engineering in advancing biocatalytic applications. |
| WOS关键词 | ASYMMETRIC-SYNTHESIS ; DEHYDROGENASE ; DEPENDENCE |
| 资助项目 | National Key Research and Devel-opment Program of China[2021YFC2102100] ; Shanghai Sailing Pro-gram[23YF1445800] ; Shenzhen Science and Technology Program[JCYJ20220818100804009] ; Guangdong Pearl River Talent Pro-gram[2021QN020121] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Chemistry |
| 语种 | 英语 |
| WOS记录号 | WOS:001631501000001 |
| 出版者 | ACADEMIC PRESS INC ELSEVIER SCIENCE |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/322157]  |
| 专题 | 中国科学院上海药物研究所 |
| 通讯作者 | Li, Hongchun; Chen, Shaoxin |
| 作者单位 | 1.China State Inst Pharmaceut Ind, Shanghai Inst Pharmaceut Ind, Natl Key Lab Lead Druggabil Res, 285 Gebaini Rd, Shanghai 201203, Peoples R China 2.Shenzhen Bay Lab, Inst Syst & Phys Biol, Gaoke Innovat Ctr, Shenzhen 518132, Peoples R China 3.Shandong Normal Univ, 88 East Wenhua Rd, Jinan 250014, Shandong, Peoples R China 4.Westlake Univ, 600 Dunyu Rd, Hangzhou 310030, Zhejiang, Peoples R China 5.Chinese Acad Sci, Zhongshan Inst Drug Discovery, Shanghai Inst Mat Med, Zhongshan 528400, Guangdong, Peoples R China |
| 推荐引用方式 GB/T 7714 | Tang, Jiawei,Zhong, Jirui,Chen, Hao,et al. Cavity engineering-driven switching of cofactor specificity in a carbonyl reductase for the synthesis of chiral aryl β-Hydroxy α-amino acid[J]. BIOORGANIC CHEMISTRY,2025,167:9. |
| APA | Tang, Jiawei.,Zhong, Jirui.,Chen, Hao.,Zhang, Luwen.,Shao, Junxi.,...&Chen, Shaoxin.(2025).Cavity engineering-driven switching of cofactor specificity in a carbonyl reductase for the synthesis of chiral aryl β-Hydroxy α-amino acid.BIOORGANIC CHEMISTRY,167,9. |
| MLA | Tang, Jiawei,et al."Cavity engineering-driven switching of cofactor specificity in a carbonyl reductase for the synthesis of chiral aryl β-Hydroxy α-amino acid".BIOORGANIC CHEMISTRY 167(2025):9. |
入库方式： OAI收割
来源：上海药物研究所
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