Zakalskiy Andriy E, Zakalska Oksana M, Rzhepetskyy Yuriy A, Potocka Natalia, Stasyk Oleh V, Horak Daniel, Gonchar Mykhailo V
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.
Protein Expr Purif. 2012 Jan;81(1):63-68. doi: 10.1016/j.pep.2011.09.001. Epub 2011 Sep 17.
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.
精氨酸酶（EC 3.5.3.1；L-精氨酸脒基水解酶）是尿素循环中的关键酶，催化精氨酸转化为鸟氨酸和尿素，这是哺乳动物肝脏中尿素形成的最后一个胞质反应。构建了能够在高效铜诱导型启动子CUP1的控制下从人肝脏中过量产生精氨酸酶I（rhARG1）的酿酒酵母重组菌株。通过用Ni-NTA琼脂糖进行金属亲和层析，从重组菌株的无细胞提取物中一步纯化出（His）6标记的rhARG1。纯化40倍的酶的最大比活性为1600 μmol min-1 mg-1蛋白质。
