Partial purification and characterization of alpha-glucosidase from Pseudomonas fluorescens W.
The alpha-glucosidase (alpha-D-glucoside glucohydrolase, EC 3.2.1.20) of Pseudomonas fluorescens W was partially purified by (NH4)2SO4 fractionation, Sephadex G-200 and DEAE-cellulose column chromatography. The enzyme showed great specificity for maltose hydrolysis, with very little action against polymeric forms. Sucrose, isomaltose, alpha-methylglucoside, and maltobionic acid were not hydrolyzed. Turanose was a strong competitive inhibitor, and glucose a weaker one. Tris (2-amino-2-hydroxymethylpropan-1:3-diol) inhibited enzyme activity significantly only at alkaline pH. Mercuric, cupric, and silver cations strongly inhibited, and EDTA (ethylenediaminetetraacetate) weakly inhibited the enzyme. The isolated enzyme was rather unstable even at 4 degrees C, and was destroyed by freezing and lyophilization. Inositol and albumin had a slightly protective effect. Sulfhydryl-binding reagents strongly inhibited the enzyme.
D004355:Drug Stability; D004492:Edetic Acid / Q000494:pharmacology; D005959:Glucosidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D007477:Ions / Q000494:pharmacology; D007700:Kinetics; D008320:Maltose; D009844:Oligosaccharides; D011551:Pseudomonas fluorescens / Q000201:enzymology*; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013696:Temperature; D014325:Tromethamine
D007477:Ions; D009844:Oligosaccharides; D013439:Sulfhydryl Reagents; D014325:Tromethamine; D008320:Maltose; D004492:Edetic Acid; D005959:Glucosidases