Loading…
A Novel Thermal Stable Carbonyl Reductase from Bacillus cereus by Gene Mining as Biocatalyst for β-Carbonyl Ester Asymmetric Reduction Reaction
Carbonyl reductase, as biocatalyst, is very important to chiral alcohols production through asymmetric reduction of carbonyl compound. A novel thermal stable carbonyl reductase from Bacillus cereus (BcCR) dependant on NADPH was obtained through a new genome mining strategy proposed in this work. By...
Saved in:
Published in: | Catalysis letters 2019-02, Vol.149 (2), p.610-618 |
---|---|
Main Authors: | , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
Summary: | Carbonyl reductase, as biocatalyst, is very important to chiral alcohols production through asymmetric reduction of carbonyl compound. A novel thermal stable carbonyl reductase from
Bacillus cereus
(BcCR) dependant on NADPH was obtained through a new genome mining strategy proposed in this work. By analyzing its amino acid sequence and structure, the BcCR should be a thermal stability and wide pH tolerance carbonyl reductase. Its gene was cloned by PCR with
B. cereus
genomic DNA as template. Its heterologous expression system,
E. coli
BL21 (DE3) plysS/pET28a-
bccr
, was constructed, and BcCR was successfully expressed. Enzymatic properties show that at 57.5 °C and pH 7.0 it can reach maximum reaction rate. Its K
m
and V
max
to ethyl 4-chloroacetoacetate is 1.85 mmol/L and 0.22 µmol/(min·mg
protein
), respectively. It can catalyze the asymmetric reduction of the β-carbonyl compound, such as ethyl 4-chloroacetoacetate to ethyl
S
-4-chloro-3-hydroxybutyrate. This paper proposes a practical method for discovery of new carbonyl reductases, and provides a novel enzyme as biocatalyst for asymmetric reduction of β-carbonyl compound. |
---|---|
ISSN: | 1011-372X 1572-879X |
DOI: | 10.1007/s10562-018-2645-4 |