Expression in Escherichia coli and characterization of an aldehyde reductase from a marine bacterium, Vibrio sp. strain XY-214
2008
Umemoto, Y.(Mie Univ., Tsu (Japan)) | Araki, T.
An aldehyde reductase gene (alrA) located between a beta-1,3-xylanase gene (txyA) and a beta-1, 3-xylosidase gene (xloA) was cloned from a marine bacterium Vibrio sp. strain XY-214. The alrA gene consisted of a 975-bp nucleotide sequence encoding a protein of 324 amino acids with a predicted molecular weight of 36,367. The deduced protein (named AlrA) showed similarities to members of the aldo-keto reductase (AKR) superfamily. The intact coding region for AlrA was subcloned into pCold TF DNA vector and expressed in Escherichia coli BL21 (DE3). Recombinant AlrA (rAlrA), which was treated with thrombin protease to remove a Trigger Factor (TF) and a six-His tag at its N-terminal, was purified with HiTrap chelating HP and Resource Q column chromatographies. AlrA showed activity toward P-nitrobenzaldehyde, but no activities were observed toward D-xylose, D-arabinose, D-glucose, D-mannose, D-fructose, and D-maltose (all at 20 mM). These results suggested that AlrA of Vibrio sp. strain XY-214 is an aldehyde reductase of the AKR superfamily.
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