EXPRESSION OF RECOMBINANT HUMAN ARGININOSUCCINATE SYNTHETASE IN  ESCHERICHIA COLI

O. I. Vovk; Y. A. Rzhepetskyy; H. G. Pereverzeva; O. V. Stasyk

doi:10.30970/sbi.0503.173

EXPRESSION OF RECOMBINANT HUMAN ARGININOSUCCINATE SYNTHETASE IN ESCHERICHIA COLI

O. I. Vovk, Y. A. Rzhepetskyy, H. G. Pereverzeva, O. V. Stasyk

DOI: http://dx.doi.org/10.30970/sbi.0503.173

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

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References

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