中国医科大学学报2013，Vol.42Issue(1)：52-55,4.
人源醛酮还原酶7A2融合蛋白的纯化及其对苯代谢产物的催化活性
Purification of Recombinant Human AKR7A2 Protein and Identification of the Catalytic Activity of AKR7A2 towards Benzene Metabolites
摘要
Abstract
Objective To investigate the catalytic activity of AKR7A2 towards certain benzene metabolites using an FPLC purified human aldo-keto reductase AKR7A2 fusion protein. Methods The plasmid containing His-AKR7A2 sequence was introduced into BL21 stain. After IPTG induction, His tagged AKR7A2 fusion protein was purified with HiTrap column and G25 Sephadex column using FPLC system. SDS-PAGE and Western blot were carried out to identify the purified AKR7A2 protein. AKR enzyme assay was applied to measure the substrate specificity of purified recombinant AKR7A2 protein towards benzaldehyde,p-benzoquinone and t,t-muconaldehyde. Results The re-combinant His-AKR7A2 fusion protein was successfully purified using two step purification with over 98% purity. The AKR7A2 fusion protein was confirmed by Western blot. Results from enzyme assay showed that AKR7A2 protein displayed mild substrate specificity towards benzaldehyde, high specificity towards t, t-muconaldehyde and low activity towards p-benzoquinone. Conclusion The recombinant human AKR7A2 was successfully purified and showed substrate specificity towards certain benzene metabolites, these findings suggest an important role of aldo-keto reductases in benzene metabolic pathways.关键词
醛酮还原酶/蛋白纯化/底物特异性Key words
aldo-keto reductase/ protein purification/ substrate specificity分类
生物科学引用本文复制引用
李丹,初阳,刘雅茹,张歧山,秦勇..人源醛酮还原酶7A2融合蛋白的纯化及其对苯代谢产物的催化活性[J].中国医科大学学报,2013,42(1):52-55,4.基金项目
留学人员科技活动择优资助项目(人社厅函[2012]258) （人社厅函[2012]258）
辽宁省科学技术计划项目(2012225021) （2012225021）
