title: Immobilization of solubilized UDP-glucuronosyltransferase from rat liver microsomes to Sepharose 4B.
abstract: A method for the covalent binding of rat liver UDP-glucuronosyltransferase to a cyanogen bromide-activated agarose matrix is described. The rat liver microsomal fraction was solubilized with 8 mM 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS); 90% of the microsomal protein was solubilized. Some 50-60% of this protein became bound covalently to the activated agarose matrix. The immobilized UPD-glucuronosyltransferase remained completely active for 50 days when stored at 4 degrees in a 20% (v/v) glycerol buffer (pH 7.4). The immobilized enzyme has a temperature optimum around 37 degrees, and a broad pH optimum (pH 5-7.4). The enzyme displayed linear kinetics over a period of 1 hr; it conjugates a large variety of substrates.
authors.affiliation: Division of Toxicology, University of Leiden, The Netherlands.
authors.forename: R A
authors.initials: RA
authors.lastname: Gilissen
authors.forename: J H
authors.initials: JH
authors.lastname: Meerman
authors.forename: G J
authors.initials: GJ
authors.lastname: Mulder
mesh_terms: D000818:Animals; D002793:Cholic Acids; D003488:Cyanogen Bromide; D004795:Enzyme Stability; D014453:Glucuronosyltransferase / Q000737:chemistry / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D012685:Sepharose; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D002793:Cholic Acids; D012685:Sepharose; D014453:Glucuronosyltransferase; D003488:Cyanogen Bromide; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate