The solubilization of a glucuronyltransferase involved in pea (Pisum sativum var. Alaska) glucuronoxylan synthesis.
A glucuronyltransferase involved in glucuronoxylan biosynthesis was obtained from the epicotyls of 1-week-old etiolated pea (Pisum sativum var. Alaska) seedlings and was solubilized in Triton X-100, a non-ionic detergent. The enzyme was inactivated by SDS and inhibited by Derriphat 160 and cholic acid. The enzyme was active in the presence of NN-dimethyldodecylanium-N-oxide, but was not solubilized by it. The stimulatory effect of UDP-D-xylose on the particulate and solubilized enzymes was the same, but the optimum Mn2+ concentration was lower for the solubilized enzyme, and the product formed by the solubilized enzyme has altered structure and solubility properties. Gel filtration of the solubilized enzyme on Sepharose CL-6B permitted partial separation of the stimulatory effect of UDP-D-xylose from the activity in the absence of UDP-D-xylose. The solubilized enzyme was more stable than the particulate enzyme and could be stored for 2 weeks at -20 degrees C without loss of activity.
Waldron
K W
KW
AFRC Institute of Food Research, Norwich, U.K.
Baydoun
E A
EA
Brett
C T
CT
D002250:Carbon Radioisotopes; D002850:Chromatography, Gel; D003902:Detergents / Q000494:pharmacology; D007887:Fabaceae / Q000201:enzymology*; D014453:Glucuronosyltransferase / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D017830:Octoxynol; D010946:Plants, Medicinal; D011092:Polyethylene Glycols / Q000494:pharmacology; D011134:Polysaccharides / Q000096:biosynthesis*; D011865:Radioisotope Dilution Technique; D014535:Uridine Diphosphate Glucuronic Acid / Q000378:metabolism; D014540:Uridine Diphosphate Xylose / Q000378:metabolism; D014990:Xylans / Q000096:biosynthesis*
D002250:Carbon Radioisotopes; D003902:Detergents; D011134:Polysaccharides; D014990:Xylans; D014535:Uridine Diphosphate Glucuronic Acid; D014540:Uridine Diphosphate Xylose; C038910:glucuronoxylan; D011092:Polyethylene Glycols; D008345:Manganese; D017830:Octoxynol; D014453:Glucuronosyltransferase