Abstract

EXPRESSION, PURIFICATION AND ACTIVITY OF RECOMBINANT HUMAN ACETALDEHYDE DEHYDROGENASE II

Kang-Jia Liu, Zhen-Ru Zhou, Yang Liu, Feng Wang and *Ren-Wang Jiang

ABSTRACT

In order to improve the expression of human acetaldehyde dehydrogenase II (ALDH2) gene in prokaryotes, the ALDH2 gene was constructed on the pET-32a (+) expression vector containing the TrxA fusion protein tag and 6×His tag, and transformed into BL21(DE3)pLysS cells. Induced by isopropylthio-β-D-galactoside (IPTG), the transformed gene was expressed in E. coli. By optimizing the expression conditions, it was determined that the optimal OD= 1.0, the final IPTG concentration = 0.5 mol/L, glucose concentration= 1% and the culture was set at 37 °C, 180 rpm/min for 16 h. The addition of the TrxA fusion protein tag can increase the soluble expression of ALDH2 in E. coli and has no significant effect on the activity of ALDH2 protein.

Keywords: Acetaldehyde dehydrogenase 2, prokaryotic expression, TrxA fusion protein tag.