Overexpression of (His)6-tagged human arginase I in Saccharomyces cerevisiae and enzyme purification using metal affinity chromatography

0364854 - ÚMCH 2012 RIV US eng J - Journal Article
Zakalskiy, A. E. - Zakalska, O. M. - Rzhepetskyy, Y. A. - Potocka, N. - Stasyk, O. V. - Horák, Daniel - Gonchar, M. V.
Overexpression of (His)6-tagged human arginase I in Saccharomyces cerevisiae and enzyme purification using metal affinity chromatography.
Protein Expression and Purification. Roč. 81, č. 1 (2012), s. 63-68. ISSN 1046-5928. E-ISSN 1096-0279
R&D Projects: GA ČR GA203/09/1242
Institutional research plan: CEZ:AV0Z40500505
Keywords : human arginase I * (His)6-tag * Saccharomyces cerevisiae
Subject RIV: CD - Macromolecular Chemistry
Impact factor: 1.429, year: 2012

Arginase (EC 3.5.3.1; L-arginine amidinohydrolase) is a key enzyme of the urea cycle that catalyses the conversion of arginine to ornithine and urea, which is the final cytosolic reaction of urea formation in the mammalian liver. The recombinant strain of the yeast Saccharomyces cerevisiae that is capable of overproducing arginase I (rhARG1) from human liver under the control of the efficient copper-inducible promoter CUP1, was constructed. The (His)6-tagged rhARG1 was purified in one step from the cell-free extract of the recombinant strain by metal-affinity chromatography with Ni–NTA agarose. The maximal specific activity of the 40-fold purified enzyme was 1600 μmol min−1 mg−1 protein.
Permanent Link: http://hdl.handle.net/11104/0200227