[Cloning, expression and the application of human, rat alcohol dehydrogenase and aldo-keto reductase].
Yao Xue Xue Bao
; 44(7): 778-84, 2009 Jul.
Article
em Zh
| MEDLINE
| ID: mdl-19806920
ABSTRACT
This study is aimed to clone and express human, rat alcohol dehydrogenase (ADH) and aldo-keto reductase. Then the enantioselective metabolism of mandelic acid (MA) was studied. Human alcohol dehydrogenase 2, rat alcohol dehydrogenase 1, human and rat aldo-keto reductase 1A1 were amplified using RT-PCR from human and rat liver samples. Then subcloned into pET-28a (+) and expressed in E. coli BL21 (DE3) stably. The protein was induced with IPTG and purified by affinity chromatography. Then the enzyme activities were measured. MA enantiomers were incubated with rat, human ADH and phenylglyoxylic acid (PGA) with AKR1A1, respectively. The metabolism was analyzed with HPLC. The proper genes were cloned and purified and proteins were obtained. All of the proteins obtained showed good activity. Stereoselective-metabolism of MA was observed in human ADH2, which favors for S-MA metabolism. The expression plasmids are constructed and the recombinant proteins are expressed successfully. The recombinant alcohol dehydrogenase and aldo-keto reductase have been employed to study MA metabolism.
Buscar no Google
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Álcool Desidrogenase
/
Proteínas Recombinantes
/
Oxirredutases do Álcool
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Yao Xue Xue Bao
Ano de publicação:
2009
Tipo de documento:
Article
País de afiliação:
China