title: Overexpression of (His)6-tagged human arginase I in Saccharomyces cerevisiae and enzyme purification using metal affinity chromatography.
abstract: 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.
authors.affiliation: Institute of Cell Biology, National Academy of Sciences of Ukraine, 14/16 Drahomanov Str., Lviv 79005, Ukraine. Electronic address: zakalska@yahoo.com.
authors.forename: Andriy E
authors.initials: AE
authors.lastname: Zakalskiy
authors.affiliation: Institute of Cell Biology, National Academy of Sciences of Ukraine, 14/16 Drahomanov Str., Lviv 79005, Ukraine. Electronic address: zakalska@yahoo.com.
authors.forename: Oksana M
authors.initials: OM
authors.lastname: Zakalska
authors.affiliation: Institute of Cell Biology, National Academy of Sciences of Ukraine, 14/16 Drahomanov Str., Lviv 79005, Ukraine. Electronic address: rzhepetskyy@cellbiol.lviv.ua.
authors.forename: Yuriy A
authors.initials: YA
authors.lastname: Rzhepetskyy
authors.affiliation: Branch Campus of the Faculty of Biotechnology, Rzeszów University, ul. Sokolowska 26, PL-36-100 Kolbuszowa, Poland. Electronic address: natpotocka@wp.pl.
authors.forename: Natalia
authors.initials: N
authors.lastname: Potocka
authors.affiliation: Institute of Cell Biology, National Academy of Sciences of Ukraine, 14/16 Drahomanov Str., Lviv 79005, Ukraine. Electronic address: stasyk@cellbiol.lviv.ua.
authors.forename: Oleh V
authors.initials: OV
authors.lastname: Stasyk
authors.affiliation: Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského Sq. 2, 162 06 Prague, Czech Republic. Electronic address: horak@imc.cas.cz.
authors.forename: Daniel
authors.initials: D
authors.lastname: Horak
authors.affiliation: Institute of Cell Biology, National Academy of Sciences of Ukraine, 14/16 Drahomanov Str., Lviv 79005, Ukraine; Branch Campus of the Faculty of Biotechnology, Rzeszów University, ul. Sokolowska 26, PL-36-100 Kolbuszowa, Poland. Electronic address: gonchar@cellbiol.lviv.ua.
authors.forename: Mykhailo V
authors.initials: MV
authors.lastname: Gonchar
mesh_terms: D001119:Arginase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D003001:Cloning, Molecular; D006639:Histidine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000201:enzymology; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives; D009842:Oligopeptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009942:Organometallic Compounds; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*
chemical_list: C471213:His-His-His-His-His-His; D009842:Oligopeptides; D009942:Organometallic Compounds; D011993:Recombinant Fusion Proteins; C088321:nickel nitrilotriacetic acid; D006639:Histidine; D001119:Arginase; D009571:Nitrilotriacetic Acid