[Purification of recombinant AKR7A5 protein and measurement of substrate specificity of AKR7A5 towards naphthoquinone and its derivatives].

AIM:To investigate the substrate specificity of mouse aldo-keto reductase AKR7A5 protein towards naphthoquinone and its derivatives. METHODS:The recombinant His-tagged AKR7A5 fusion protein in E.coli BL21pLysS cell strain was induced by IPTG and purified using FPLC system through HiTrap affinity column. The purified recombinant AKR7A5 protein was confirmed by SDS-PAGE and Western blotting. AKR enzyme assay was applied to measure the substrate specificity of recombinant AKR7A5 protein towards naphthoquinone and its derivatives. RESULTS:Recombinant His-AKR7A5 was successfully purified as confirmed by SDS-PAGE and Western blotting. AKR enzyme assay indicated that the recombinant AKR7A5 protein exhibited mild substrate specificity towards lawsone and low specificity towards juglone and vitamine K3, but no activity towards 1, 4-naphthoquinone. CONCLUSION:AKR7A5 has selective substrate specificity towards naphthoquinone derivatives, suggesting that the aldo-keto reductase could play an important role in metabolism of certain naphthoquinone derivatives.

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