Xanthine oxidase and aldehyde oxidase: a simple procedure for the simultaneous purification from rat liver

Archives of Biochemistry and Biophysics
L Maia, L Mira

Abstract

Aldehyde oxidase (AO) and xanthine oxidase (XO) are cytosolic enzymes that have been involved in some pathological conditions and play an important role in the biotransformation of drugs and xenobiotics. The increasing interest in these enzymes demands for a simple and rapid procedure for their purification. This paper describes for the first time a method that allows simultaneous purification of both enzymes from the same batch of rat livers. It involves few steps, is reproducible and offers high enzyme yields with high specific activities. The rat liver homogenate was fractionated by heat denaturation and by ammonium sulphate precipitation to give a crude extract containing both enzymes. This extract was chromatographed on an Hydroxyapatite column that completely separated AO from XO. Further purification of XO by anion exchange chromatography on a Q-Sepharose Fast Flow column resulted in a highly purified (1200-fold) preparation, with a specific activity of 3.64 U/mg and with a 20% yield. AO was purified about 1000-fold at a yield of 15%, with a specific activity of 3.48 U/mg, by affinity chromatography on Benzamidine-Sepharose 6B. The purified enzymes gave single bands of approximately 300 kDa on a polyacrylamide gel gradie...Continue Reading

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Citations

Mar 7, 2013·Toxicological Sciences : an Official Journal of the Society of Toxicology·Tami L Swenson, John E Casida
Jan 6, 2006·Chemical & Pharmaceutical Bulletin·Saieed PirouzpanahHamidi Aliasghar
Aug 5, 2014·Journal of Enzyme Inhibition and Medicinal Chemistry·Serap Beyaztaş, Oktay Arslan
May 17, 2006·Drug Metabolism and Pharmacokinetics·Shigeyuki KitamuraShigeru Ohta
Mar 14, 2007·Archives of Biochemistry and Biophysics·Tapan Kumar KunduJay L Zweier
Apr 4, 2014·Chemical Reviews·Luisa B Maia, José J G Moura
Dec 9, 2003·FEMS Microbiology Letters·Hiroyuki UchidaTakayuki Uwajima
Apr 19, 2007·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Luisa MaiaLurdes Mira
Nov 16, 2007·Chemical Research in Toxicology·Ryan A DickJohn E Casida
Nov 14, 2003·Toxicology and Applied Pharmacology·H Vasken AposhianMichael L Wollenberg
Feb 22, 2005·Chemical Research in Toxicology·Ryan A DickJohn E Casida
Nov 15, 2011·Poultry Science·A SaengtienchaiM Ishizuka

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