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Promotion of the Asymmetric Reduction of Prochiral Ketone with Recombinant E. coli Through Strengthening Intracellular NADPH Supply by Modifying EMP and Introducing NAD Kinase
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Promotion of the Asymmetric Reduction of Prochiral Ketone with Recombinant E. coli Through Strengthening Intracellular NADPH Supply by Modifying EMP and Introducing NAD Kinase
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Journal Article
Promotion of the Asymmetric Reduction of Prochiral Ketone with Recombinant E. coli Through Strengthening Intracellular NADPH Supply by Modifying EMP and Introducing NAD Kinase
2021
Overview
The intracellular NADPH insufficient supply is the main bottleneck to the synthesis of chiral alcohols by asymmetric reduction with whole-cell catalysis. Herein, we provide a novel strategy to strengthen intracellular NADPH supply through introducing an NADP
+
-dependent glyceraldehyde 3-phosphate dehydrogenase (
gap
B from
Bacillus subtilis
168) into the Embden-Meyerhof pathway and a NAD kinase (
yfj
B from
E. coli
MG1655) to further enhance the NADP(H) pool. A recombinant
E. coli
(
E. coli
BL21 (DE3)/pETDuet-1-
gap
B-
yue
D&pET28a-
yfj
B) was constructed to co-express
gap
B and
yfj
B with a carbonyl reductase gene
yue
D together. The result showed that the intracellular NADPH amount increased by 134.4% with the strategy. To the model reaction (asymmetric reduction of acetophenone to
S
-phenyl ethanol), the yield was 3.7-fold with this strategy compared to the control. This provides a technological route for strengthening the intracellular NADPH supply in
E. coli
for biocatalysis and biosynthesis.
Graphic Abstract
Publisher
Springer US,Springer,Springer Nature B.V
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