Partial purification of (1,3)-beta-glucan synthase from Candida albicans.
(1,3)-beta-Glucan synthase from Candida albicans was solubilized from microsomal membranes using the detergent 3-[(3-cholamidopropyl) dimethylammonio]-1-propane sulfonate (Chaps). Effective solubilization was dependent upon the strain and the method used to detect enzyme activity. The solubilized enzyme was purified over 765-fold using a modified product entrapment technique. Bovine serum albumin, an activator of glucan synthase, precipitated proteins during product entrapment and was replaced with BSA immobilized on agarose beads. SDS-PAGE analysis revealed a prominent 187-kDa band present in the product entrapped pellet as well as several additional polypeptides at 227, and 187, 182 and 39 kDa which were not prevalent in crude preparations.
Frost
D
D
Department 47M, Abbott Laboratories, Abbott Park, IL 60064-3500, USA. david.frost@abbott.com
Brandt
K
K
Estill
C
C
Goldman
R
R
D002176:Candida albicans / Q000201:enzymology*; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D005964:Glucosyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D008565:Membrane Proteins; D008861:Microsomes / Q000201:enzymology; D029702:Schizosaccharomyces pombe Proteins; D012995:Solubility
D003902:Detergents; D008565:Membrane Proteins; D029702:Schizosaccharomyces pombe Proteins; D005964:Glucosyltransferases; C020570:1,3-beta-glucan synthase