Affiliations: [a] West China School of Preclinical and Forensic Medicine, Sichuan University, Sichuan, China | [b] Institute of Laboratory Medicine, Beihua University, Jilin, China | [c] School of Chemical Engineering, Sichuan University, Chengdu, Sichuan, China
Correspondence:
[*]
Corresponding author: Xiufeng Gao, West China School of Preclinical and Forensic Medicine, Sichuan University, Sichuan, China. Tel.: +86 28 85503204; Fax: +86 28 85503204;beihuan2010@163.com
Abstract:
BACKGROUND: For its function of catalysing the reaction of aldehyde
to acetic acid, ALDH could be applied to antialcoholismic drug development.
However, both of the production and activity of natural ALDH are too low to
meet the demand.
OBJECTIVE: To empirically explore whether aldh gene in Bacillus halodurans XJU-1 could
be highly expressed in E.coli BL21(DE3) and to investigate the purification of
recombinant protein ALDH.
METHODS: The aldh gene in Bacillus halodurans XJU-1 was cloned into prokaryotic
expression vector pET30b(+), then transformed into E.coli BL21(DE3) to induce
expression of ALDH. Immobilized metal ion affinity chromatography (IMAC) was
applied in the separation and purification for ALDH proteins.
RESULTS: The recombinant vector for aldh gene was constructed and
expressed in BL21(DE3) successfully. The ALDH recombinant protein was
purified by ammonium sulfate precipitation and IMAC, and obtained 30.1%
of the recovery with a purification factor 8.81.
CONCLUSIONS: The value of these findings, as well as wider
implications for increasing the yield and the activity of ALDH, is
meaningful.
