2. Academic Validation
  3. Extensive gene mining and facile engineering of a novel carbonyl reductase for asymmetric synthesis of anti-aging (S)-Pro-Xylane from d-xylose

Extensive gene mining and facile engineering of a novel carbonyl reductase for asymmetric synthesis of anti-aging (S)-Pro-Xylane from d-xylose

  • Int J Biol Macromol. 2025 May;305(Pt 2):140976. doi: 10.1016/j.ijbiomac.2025.140976.
Zhe Dou 1,  Pei-Ru Hu 2,  Lin-Lin Song 2,  Yi-Cheng Wang 2,  Hao-Nan Tang 2,  Zhen-Qi Weng 3,  Ya-Jun Wang 4,  Yuan-Qiang Cai 3
Affiliations

Affiliations

  • 1 College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, PR China; Key Laboratory of Pharmaceutical Engineering of Zhejiang Province, Zhejiang University of Technology, Deqing, Zhejiang 313200, PR China; Key Laboratory for Green Pharmaceutical Technology and Equipment (Zhejiang University of Technology) of Ministry of Education, Deqing, Zhejiang 313200, PR China.
  • 2 College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, PR China; Key Laboratory of Pharmaceutical Engineering of Zhejiang Province, Zhejiang University of Technology, Deqing, Zhejiang 313200, PR China; Key Laboratory for Green Pharmaceutical Technology and Equipment (Zhejiang University of Technology) of Ministry of Education, Deqing, Zhejiang 313200, PR China.
  • 3 College of Civil Engineering, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, PR China.
  • 4 College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, PR China; Key Laboratory of Pharmaceutical Engineering of Zhejiang Province, Zhejiang University of Technology, Deqing, Zhejiang 313200, PR China; Key Laboratory for Green Pharmaceutical Technology and Equipment (Zhejiang University of Technology) of Ministry of Education, Deqing, Zhejiang 313200, PR China. Electronic address: wangyj@zjut.edu.cn.
PMID: 39952530 DOI: 10.1016/j.ijbiomac.2025.140976
Abstract

Carbonyl reductases (CRs) are promising biological macromolecules for the asymmetric synthesis of chiral secondary alcohols. A novel CR was evolved for efficient conversion of d-xylose into Anti-aging (S)-Pro-Xylane. Through extensive gene mining, an active carbonyl reductase ZmCR was identified from Zygofabospora marxiana with high reductive activity towards d-xylose-derived 1-C-(β-d-xylopyranosyl)-acetone (XPSA) and in situ cofactor regeneration capability. Using a three-round of concise engineering strategy, a hextuple mutant C85V/V128C/S129A/V161S/Y166H/T217A (M6) was identified from merely 157 mutants with specific activity of 46.00 U·mg-1, about 306 folds of WT. The Shannon-Wiener index of M6 was 2.21, much higher than WT, indicating its extended substrate scope. M6 could efficiently catalyze the asymmetric reduction of 190 g·L-1 XPSA into (S)-XPSP. The kcat/KMXPSA of M6 was 203.84 s-1·mM-1, about 910 folds of WT. More non-bonded interactions were monitored in M6, contributing to the stabilization of reactive catalytic conformation. This study provides a promising biocatalyst for the chemoenzymatic synthesis of value-added Anti-aging (S)-Pro-Xylane from lignocellulosic d-xylose.

Keywords

(S)-Pro-Xylane; Carbonyl reductase; Scanning mutagenesis; d-Xylose; β-C-xyloside.

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