A multi-enzyme cascade process for the preparation of L-phosphinothricin.
- Author:
Manman WANG
1
;
Yu YANG
1
;
Xianbing SONG
1
;
Xiaolian LI
1
;
Binchun LI
2
;
Ziqiang WANG
1
Author Information
- Publication Type:Journal Article
- Keywords: D-amino acid transaminase; L-phosphinothricin; coenzyme regeneration; glutamate dehydrogenase; multi-enzyme cascade reaction
- MeSH: Escherichia coli/enzymology*; Aminobutyrates/metabolism*; Glutamate Dehydrogenase/biosynthesis*; Glucose 1-Dehydrogenase/biosynthesis*; Herbicides/metabolism*; Multienzyme Complexes/metabolism*; Transaminases/metabolism*; Phosphinic Acids/metabolism*
- From: Chinese Journal of Biotechnology 2025;41(9):3589-3603
- CountryChina
- Language:Chinese
- Abstract: L-phosphinothricin (L-PPT) is an efficient broad-spectrum herbicide. To realize the multi-enzyme catalytic preparation of L-PPT, we constructed an engineered strain Escherichia coli YM-1 for efficient expression of D-amino acid transaminase, which could catalyze the generation of the intermediate 2-oxo-4-[(hydroxymethylphosphonyl)] butyric acid (PPO) from D-phosphinothricin (D-PPT). In addition, E. coli pLS was constructed to co-express glutamate dehydrogenase and glucose dehydrogenase, which not only catalyzed the generation of L-PPT from PPO but also regenerated the coenzyme nicotinamide adenine dinucleotide phosphate (NADPH). A fed-batch fermentation process was then established for E. coli YM-1 and pLS, and the apparent activities of D-amino acid transaminase and glutamate dehydrogenase were increased by 22.68% and 100.82%, respectively, compared with those in shake flasks. The process parameters were optimized for the catalytic preparation of L-PPT by whole-cell cascade of E. coli YM-1 and pLS with D, L-PPT as the substrate. After reaction for 8 h, 91.36% conversion of D-PPT was achieved, and the enantiomeric excess of L-PPT reached 90.22%. The findings underpin the industrial production of L-PPT.
