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中国生物工程杂志

China Biotechnology
China Biotechnology  2017, Vol. 37 Issue (1): 64-70    DOI: 10.13523/j.cb.20170110
    
One-pot Enzymatic Synthesis of L-2-Aminobutyric Acid Coupling with a NADH Regeneration System Based on Ketoreductase
LIU Shan, LI Yuan, ZHU Jun
Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China
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Abstract  

A process of one-pot enzymatic synthesis of L-2-aminobutyric acid was studied. The ilva gene encoding L-threonine deaminase (L-TD) from E. coli, leudh gene encoding leucine dehydrogenase (LeuDH) from Geobacillus stearothermophilus and kred gene encoding ketoreductase (KRED) from Novosphingobium aromaticivorans were cloned into pET-21a(+), then introduced into E. coli BL21(DE3) respectively. All of the three enzymes were overexpressed in soluble forms. L-2-aminobutyric acid was efficiently synthesized with low-cost L-threonine as substrate by the recombinant L-TD and LeuDH coupling with a NADH regeneration system based on KRED. Effects of pH and the concentration of L-threonine and isopropanol on the yield of L-2-aminobutyric acid were investigated. After 20h at the optimal pH 7.5~8.0, the yield of L-2-aminobutyric acid could achieve 99%, with a concentration of 43g/L, when using 50g/L L-threonine, 5% isopropanol, 0.5g/L NAD+, 0.6g/L L-TD, 2g/L LeuDH and 2g/L KRED. A new thought for the production of L-2-aminobutyric acid was provided.



Key wordsEnzymatic synthesis      L-2-aminobutyric acid      NADH regeneration      One-pot     
Received: 04 July 2016      Published: 25 January 2017
ZTFLH:  Q814.9  
Cite this article:

LIU Shan, LI Yuan, ZHU Jun. One-pot Enzymatic Synthesis of L-2-Aminobutyric Acid Coupling with a NADH Regeneration System Based on Ketoreductase. China Biotechnology, 2017, 37(1): 64-70.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20170110     OR     https://manu60.magtech.com.cn/biotech/Y2017/V37/I1/64

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