Production of L-2-aminobutyric acid from L-threonine using a trienzyme cascade.

Yan FU; Junxuan Zhang; Xuerong FU; Yuchen Xie; Hongyu Ren; Jia Liu; Xiulai Chen; Liming Liu

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Production of L-2-aminobutyric acid from L-threonine using a trienzyme cascade.

Author: Yan FU ¹ ; Junxuan ZHANG ¹ ; Xuerong FU ¹ ; Yuchen XIE ¹ ; Hongyu REN ¹ ; Jia LIU ¹ ; Xiulai CHEN ¹ ; Liming LIU ¹
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1. State Key Laboratory of Food Science & Technology, Jiangnan University, Wuxi 214122, Jiangsu, China.
Publication Type:Journal Article
Keywords: L-2-aminobutyric acid; L-threonine; co-expressed recombinant bacteria; trienzyme cascade; whole-cell catalyst
MeSH: Aminobutyrates; chemical synthesis; Bacillus thuringiensis; enzymology; Candida; enzymology; Escherichia coli; enzymology; Formate Dehydrogenases; metabolism; Leucine Dehydrogenase; metabolism; Threonine; metabolism; Threonine Dehydratase; metabolism
From: Chinese Journal of Biotechnology 2020;36(4):782-791
CountryChina
Language:Chinese
Abstract: L-2-aminobutyric acid (L-ABA) is an important chemical raw material and chiral pharmaceutical intermediate. The aim of this study was to develop an efficient method for L-ABA production from L-threonine using a trienzyme cascade route with Threonine deaminase (TD) from Escherichia. coli, Leucine dehydrogenase (LDH) from Bacillus thuringiensis and Formate dehydrogenase (FDH) from Candida boidinii. In order to simplify the production process, the activity ratio of TD, LDH and FDH was 1:1:0.2 after combining different activity ratios in the system in vitro. The above ratio was achieved in the recombinant strain E. coli 3FT+L. Moreover, the transformation conditions were optimized. Finally, we achieved L-ABA production of 68.5 g/L with a conversion rate of 99.0% for 12 h in a 30-L bioreactor by whole-cell catalyst. The environmentally safe and efficient process route represents a promising strategy for large-scale L-ABA production in the future.