Expression of recombinant human argininosuccinate synthetase in Escherichia coli

Abstract

Escherichia coli strain BL21(DE3)/pET42a/ASS – an efficient producer of recombinant human argininosuccinate synthetase (rhASS) – was constructed, and preparations of purified rhASS were obtained using His-tag affinity chromatography. The effect of specific inhibitor, α-methyl-DL-aspartate, and nitric oxide donor, sodium nitroprusside, on the ASS specific activity was evaluated with purified rhASS protein and in mouse liver lysates. The developed expression platform is a useful tool in search for new ASS inhibitors efficient under in vitro and in vivo conditions.

Keywords

• argininosuccinate synthetase
• arginine biosynthesis
• recombinant protein expression
• affinity purification