Solubiliztion and Partial Characterization of Alkylglycerol Monooxygenase from the Liver Mkcosomes

Abstract

The enzyme alkylgloycerol monooxygenase wa slubuilized ith 2% of Triton X‐100 and partially pufified approximatelkyt to 83–fold with a 36% yield after a procedure which included 6‐aminohexyl‐Sephaorse and DEAE cellulose column chromatography, followed by a surcros edensity gradient centrifugation. The moleucular weight of the native form was estimated to be appromiately 400000 as judged by Sepharose 6B column chromatography in the presence of Triton X‐100. The enzyme had a pH otimum nbear 8.5 and a Km of 0.66 for 1‐O‐hexadecylglycerol. intiaal velocity excepyt to maintiain it for a much longer longer period of the time. The purified enzyme required abslutely both molecular oxygen and reduced pteridine as an electron donor. Furthermore, reduced glutathoione glutathione an phospholipids wer necessary for expression of full enzyme activity. N‐Ethylmaleimide and p‐Chloromercuribenzoate singnificantly inhibited the enzyme activity, suggesting that alkylycerol monoxxygense is a ‐SH enzyme. Copyright © 1983, Wiley Blackwell. All rights reserved