Efficient synthesis of L-phosphinothricin using a novel aminoacylase mined from Stenotrophomonas maltophilia

Cheng-Hao Cao; Feng Cheng; Ya-Ping Xue; Yu-Guo Zheng

doi:10.1016/j.enzmictec.2019.109493

Efficient synthesis of L-phosphinothricin using a novel aminoacylase mined from Stenotrophomonas maltophilia

Enzyme Microb Technol. 2020 Apr:135:109493. doi: 10.1016/j.enzmictec.2019.109493. Epub 2019 Dec 12.

Authors

Cheng-Hao Cao¹, Feng Cheng¹, Ya-Ping Xue², Yu-Guo Zheng¹

Affiliations

¹ Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China; Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou 310014, China; National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou 310014, China.
² Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China; Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou 310014, China; National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou 310014, China. Electronic address: xyp@zjut.edu.cn.

PMID: 32146938
DOI: 10.1016/j.enzmictec.2019.109493

Abstract

L-phosphinothricin (L-PPT) is a competitive and environmentally friendly herbicide. To develop an efficient approach for synthesis of l-PPT, a kinetic resolution route with a novel aminoacylase (SmAcy) mined from Stenotrophomonas maltophilia using N-acetyl-PPT as a substrate was first constructed. This SmAcy exhibited high hydrolytic activity and excellent enantioselectivity (E > 200) toward N-acetyl-PPT. Optically pure l-PPT (> 99.9 % ee_p) was acquired with high conversion (> 49 %) within 4 h by the whole cells. On the basis of the docking analysis, a main reason for high enantioselectivity (E > 200) of SmAcy towards l-enantiomer would be that the D-N-acetyl-PPT cannot interact with the key general acid-base residue and the metal ions. A low-cost and simple preparation process of the substrate from commercially available racemic PPT for production of L-PPT was provided. A chemical racemization method of the unreacted D-enantiomer of substrate was also provided to recycle the unwanted substrate enantiomer. This study provides a potential route for the industrial production of L-PPT.

Keywords: Aminoacylase; Enzyme catalysis; Kinetic resolution; L-phosphinothricin; Racemization.

MeSH terms

Amidohydrolases / chemistry*
Aminobutyrates / chemistry*
Bacterial Proteins / chemistry*
Biocatalysis
Herbicides / chemistry*
Kinetics
Stenotrophomonas maltophilia / chemistry
Stenotrophomonas maltophilia / enzymology*
Stereoisomerism
Substrate Specificity

Substances

Aminobutyrates
Bacterial Proteins
Herbicides
phosphinothricin
Amidohydrolases
aminoacylase I