|
KMID : 0545120100200091300
|
|
Journal of Microbiology and Biotechnology
2010 Volume.20 No. 9 p.1300 ~ p.1306
|
|
|
Enantioselective bioconversion using Escherichia coli cells expressing Saccharomyces cerevisiae reductase and Bacillus subtilis glucose dehydrogenase
|
|
Park Hyun-Joo
Jung Ji-Hye
Choi Hye-Jeong
Uhm Ki-Nam
Kim Hyung-Kwoun
|
|
|
Abstract
|
|
|
|
Ethyl (R, S)-4-chloro-3-hydroxybutanoate (ECHB) is a useful chiral building block for the synthesis of L-carnitine and hypercholesterolemia drugs. The yeast reductase, YOL151W (GenBank locus tag), exhibits an enantioselective reduction activity, converting ethyl-4-chlorooxobutanoate (ECOB) exclusively into (R)-ECHB. YOL151W was generated in Escherichia coli cells and purified via Ni- NTA and desalting column chromatography. It evidenced an optimum temperature of 45oC and an optimum pH of 6.5-7.5. Bacillus subtilis glucose dehydrogenase (GDH) was also expressed in Escherichia coli, and was used for the recycling of NADPH, required for the reduction reaction. Thereafter, Escherichia coli cells co-expressing YOL151W and GDH were constructed. After permeablization treatment, the Escherichia coli whole cells were utilized for ECHB synthesis. Through the use of this system, the 30 mM ECOB substrate could be converted to (R)-ECHB.
|
|
|
KEYWORD
|
|
|
Reductase, Whole cell bioconversion, Chiral intermediate, Enantioselectivity, Coupling reaction
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|