Vitamin A and carotenoids. The enzymic conversion of beta-carotene into retinal in hog intestinal mucosa.
The conversion of beta-carotene into retinal was studied in vitro with enzyme preparations from homogenates of hog intestinal mucosa. The hog mucosal enzyme was purified about 27-fold by precipitation with ammonium sulphate, chromatography on DEAE-Sephadex and gel filtration on Sephadex G-200. The reaction displayed a narrow optimum pH range (approx. 7.8-8.2). The enzyme was stimulated strongly by the addition of thiols, and was inhibited by thiol inhibitors and by the chelating agents alphaalpha'-bipyridyl and o-phenanthroline. The reaction required the addition of an appropriate detergent (or bile salt); maximal activity was obtained by addition of an appropriate combination of detergents and lipid (specifically Tween 40, sodium glycocholate and sphingomyelin). The reaction displayed Michaelis kinetics with K(m)1.3x10(-6)m and V(max.)1.1nmole of retinal formed/hr. (for 0.7mg. of enzyme protein). The properties of the hog enzyme are similar to those previously reported for a less purified rat enzyme preparation.
D000818:Animals; D001647:Bile Acids and Salts; D002338:Carotenoids / Q000378:metabolism*; D002845:Chromatography; D002850:Chromatography, Gel; D003902:Detergents; D004791:Enzyme Inhibitors; D004798:Enzymes / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007413:Intestinal Mucosa / Q000201:enzymology / Q000378:metabolism*; D007700:Kinetics; D013438:Sulfhydryl Compounds; D013552:Swine; D014801:Vitamin A / Q000096:biosynthesis*
D001647:Bile Acids and Salts; D003902:Detergents; D004791:Enzyme Inhibitors; D004798:Enzymes; D013438:Sulfhydryl Compounds; D014801:Vitamin A; D002338:Carotenoids