Production of phenylpyruvic acid by engineered L-amino acid deaminase from Proteus mirabilis.

Jia Liu, Jianmin Liu, Bin Yang, Cong Gao, Wei Song, Guipeng Hu, Liming Liu, Jing Wu

Journal: Biotechnology letters 2022;44(5-6):635-642

PMID: 35429303

Abstract

OBJECTIVES

This study aimed to develop an efficient enzymatic strategy for the industrial production of phenylpyruvate (PPA) from L-phenylpyruvic acid (L-Phe).

RESULTS

L-amino acid deaminase from Proteus mirabilis was expressed in Escherichia coli BL21 (DE3) and modified to release product inhibition by employing conformational dynamics engineering. Based on structural analysis, two residues (E145/L341) were identified for reducing interactions between the product and enzyme and increasing flexibility of the protein, thereby facilitating the product release. The mutant M2 exhibited a 3.84-fold reduction in product inhibition and a 1.35-fold increase in catalytic efficiency in comparison to the wild type. Finally, 81.2 g/L PPA production with a conversion of 99.6% was obtained in a 5-L bioreactor.

CONCLUSIONS

The engineered catalyst can significantly reduce product inhibition and facilitate the effective industrial synthesis of PPA.

Address: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.; International Joint Laboratory on Food Safety, Jiangnan University, Wuxi, 214122, China.; Shandong Huishilai Biotechnology Co., Ltd, Jinan, 250098, China.; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, China.; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, China. [email protected].

DOI: 10.1007/s10529-022-03245-y

Link outs

Subscription / membership required

Full Text Sources:

https://doi.org/10.1007/s10529-022-03245-y

MeSH Terms: Amino Acids, Escherichia coli, Phenylpyruvic Acids, Proteus mirabilis