title: Purification and characterization of three extracellular protopectinases with polygalacturonase activities from Trichosporon penicillatum.
abstract: In a culture filtrate of Trichosporan penicillatum B2, which is a gamma-ray irradiation mutant induced from T. penicillatum SNO3, we found three kinds of pectin-releasing enzymes, protopectinases SE1, SE2, and SE3, that have endo-polygalacturonase activity. These enzymes were purified to homogeneity with cation-exchange and size exclusion chromatographies. The major PPase in the culture filtrate was PPase SE1, which accounted for 75% of total activities in the culture filtrate, and the two others were 0.15% (PPase SE2) and 0.007% (PPase SE3). Their molecular masses were approximately 41, 41, and 42 kDa on SDS-PAGE, respectively. They had similar enzymatic properties but different PPase activity and pH- and thermo-stability. Antibody against PPase S, which is produced by strain SNO3, inhibited the activities of PPase SE1, SE2, and SE3. However PPase SE1 was completely inhibited by treatment with the anti-PPase S antibody, but the activities of PPases SE2 and SE3 remained at 20 and 50% of the original activity, respectively.
authors.affiliation: Department of Applied Biochemistry, College of Agriculture, Osaka Prefecture University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Iguchi
authors.forename: M
authors.initials: M
authors.lastname: Kishida
authors.forename: T
authors.initials: T
authors.lastname: Sakai
mesh_terms: D000595:Amino Acid Sequence; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D007527:Isoenzymes / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D011096:Polygalacturonase / Q000302:isolation & purification*; D014250:Trichosporon / Q000201:enzymology*
chemical_list: D007527:Isoenzymes; D006026:Glycoside Hydrolases; C083413:protopectinase; D011096:Polygalacturonase