Purification of Alcohol Dehydrogenase Enzyme from Chicken Liver and Immobilization Onto Florisil
Havva Ersoz
Faculty of Art and Science, Department of Chemistry (Biochemistry Division), University of Cukurova, 01330, Balcali, Adana, Turkey.
Nuri Gulesci *
Department of Chemistry and Chemical Processing Technologies, Gumushane Vocational School, Gumushane University, 29100 Gumushane, Turkey.
Ramazan Bilgin
Faculty of Art and Science, Department of Chemistry (Biochemistry Division), University of Cukurova, 01330, Balcali, Adana, Turkey.
*Author to whom correspondence should be addressed.
Abstract
The enzyme alcohol dehydrogenase (ADH) is a dimeric enzyme in which each of its subunits has a Zn2+ metal-containing catalytic domain and a cofactor binding domain. This enzyme converts alcohol into an aldehyde. In this article, the activity of the enzyme was investigated by applying the immobilization process directly to the alcohol dehydrogenase enzyme purified and activated florisil from the chicken liver. For this purpose, homogenization of chicken liver was achieved and its supernatants were separated by applying the ultracentrifugation process to the resulting homogenate. Then, % ammonium precipitation, dialysis, and ion exchange chromatography processes were performed, respectively. As a result of these processes, the hepatic alcohol dehydrogenase was purified 150.3 times compared to the coarse homogenate, and the specific activity of the enzyme was determined to be 0.631 U/mg protein. The activity of the enzyme directly immobilized was found to be 0.034 U/mg protein.
Keywords: Alcohol dehydrogenase, florisil, liver, ion exchange chromatography