Formation, separation and characterization of three beta-1,3-glucanases from Sclerotium glucanicum.
The appearance of beta-1,3-glucanases in supernatants of Sclerotium glucanicum cultures was followed by SDS-PAGE and shown to be dependent on cultivation time. Three beta-1,3-glucanases were isolated and purified. Glucanase I and III appeared homogeneous on SDS-PAGE with molecular masses of 85 and 33.5 kDa, respectively. Enzyme I was an endo-splitting beta-1,3-glucanase. In hydrolyzing laminarin it released glucose, laminaritriose and laminaribiose as major endproducts and smaller amounts of higher oligosaccharides. Enzyme III was an exo-beta-1,3-glucanase removing glucose from laminarin and gentiobiose and glucose from scleroglucan. For laminarin as substrate the Km of enzyme I and III was 2.5 and 3.33 mg/ml, respectively. Enzyme II was only partially purified and found to be also an exo-beta-1,3-glucanase, releasing glucose as the only hydrolysis product from laminarin. It did not attack scleroglucan. Its molecular weight was determined to be 78 kDa. Optimum pH and temperature of the three enzymes were determined. The three activities were significantly inhibited by 1 mM Hg2+.
Rapp
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Institut für Biochemie und Biotechnologie, Technischen Universität Braunschweig, Germany.
D004591:Electrophoresis, Polyacrylamide Gel; D005658:Fungi / Q000201:enzymology*; D043326:Glucan 1,3-beta-Glucosidase; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D005936:Glucans / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D011134:Polysaccharides / Q000378:metabolism; D013379:Substrate Specificity; D001617:beta-Glucosidase / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*
D005936:Glucans; D011134:Polysaccharides; C045938:scleroglucan; C008247:laminaran; D001617:beta-Glucosidase; D004693:Glucan Endo-1,3-beta-D-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase