title: Penicillium solitum produces a polygalacturonase isozyme in decayed Anjou pear fruit capable of macerating host tissue in vitro.
abstract: A polygalacturonase (PG) isozyme was isolated from Penicillium solitum-decayed Anjou pear fruit and purified to homogeneity with a multistep process. Both gel filtration and cation exchange chromatography revealed a single PG activity peak, and analysis of the purified protein showed a single band with a molecular mass of 43 kDa, which is of fungal origin. The purified enzyme was active from pH 3.5-6, with an optimum at pH 4.5. PG activity was detectable 0-70 C with 50 C maximum. The purified isozyme was inhibited by the divalent cations Ca(2+), Mg(2+), Mn(2+) and Fe(2+) and analysis of enzymatic hydrolysis products revealed polygalacturonic acid monomers and oligomers. The purified enzyme has an isoelectric point of 5.3 and is not associated with a glycosylated protein. The PG isozyme macerated fruit tissue plugs in vitro and produced ~1.2-fold more soluble polyuronides from pear than from apple tissue, which further substantiates the role of PG in postharvest decay. Data from this study show for the first time that the purified PG produced in decayed Anjou pear by P. solitum, a weakly virulent fungus, is different from that PG produced by the same fungus in decayed apple.
authors.affiliation: USDA-ARS, Food Quality Laboratory, Beltsville, MD 20705, USA. wayne.jurick@ars.usda.gov
authors.forename: Wayne M
authors.initials: WM
authors.lastname: Jurick
authors.forename: Ivana
authors.initials: I
authors.lastname: Vico
authors.forename: Verneta L
authors.initials: VL
authors.lastname: Gaskins
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Whitaker
authors.forename: Wesley M
authors.initials: WM
authors.lastname: Garrett
authors.forename: Wojciech J
authors.initials: WJ
authors.lastname: Janisiewicz
authors.forename: William S
authors.initials: WS
authors.lastname: Conway
mesh_terms: D002413:Cations, Divalent / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005638:Fruit / Q000382:microbiology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007526:Isoelectric Point; D007527:Isoenzymes; D007700:Kinetics; D027845:Malus / Q000382:microbiology*; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology*; D010935:Plant Diseases / Q000382:microbiology*; D011096:Polygalacturonase / Q000737:chemistry / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism; D031989:Pyrus / Q000382:microbiology*; D013696:Temperature; D014574:Uronic Acids / Q000032:analysis / Q000378:metabolism
chemical_list: D002413:Cations, Divalent; D007527:Isoenzymes; D014574:Uronic Acids; C018402:traumacel; D011096:Polygalacturonase