Efficient production of recombinant aldehyde reductase and its application for asymmetric reduction of ethyl 4-chloro-3-oxobutanoate to ethyl (R)-4-chloro-3-hydroxybutanoate.
An NADPH-dependent aldehyde reductase (ALR, EC1.1.1.2) gene is cloned from Sporobolomyces salmonicolor ZJUB 105, and inserted into plasmid pQE30 to construct the expression plasmid (pQE30-ALR). A variety of E. coli strains were employed as hosts to obtain transformants with pQE30-ALR, respectively. Among these different types of transformants, the highest enzyme activity of ALR can be produced with E. coli M15 (pQE30-ALR). The bioactivity of ALR could be further improved significantly by the optimization of induction conditions. The results showed that the enzyme activity of ALR reached 6.48 U/mg protein, which is fifteen times higher than that of S. salmonicolor ZJUB 105. This recombinant strain was applied to the asymmetric reduction of ethyl 4-chloro-3-oxobutanoate (COBE) to ethyl (R)-4-chloro-3- hydroxybutanoate (CHBE). The results showed that the yield and optical purity of (R)-CHBE reached 98.5% and 99% e.e. (enantiomeric excess), respectively.
Jing
Keju
K
Institute of Bioengineering, Department of Chemical Engineering and Bioengineering, Zhejiang University, Hangzhou, PR China.
Xu
Zhinan
Z
Liu
Ying
Y
Jiang
Xiaoxia
X
Peng
Li
L
Cen
Peilin
P
D000090:Acetoacetates / Q000378:metabolism*; D000449:Aldehyde Reductase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D001487:Basidiomycota / Q000201:enzymology* / Q000235:genetics; D002087:Butyrates / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D014169:Transformation, Bacterial
D000090:Acetoacetates; D002087:Butyrates; D005656:Fungal Proteins; D011994:Recombinant Proteins; C121244:ethyl 4-chloro-3-hydroxybutanoate; C111982:ethyl 4-chloro-3-oxobutanoate; D000449:Aldehyde Reductase