title: Properties of a polygalacturonase-inhibiting protein isolated from 'Oroblanco' grapefruit.
abstract: Polygalacturonase inhibiting protein (PGIP) was extracted from 'Oroblanco' grapefruit type (triploid pummelo-grapefruit) albedo tissue, purified and partially characterized. Extraction was carried out at 4 degrees C with a high ionic strength extraction buffer. After dialysis and concentration by ultrafiltration the extract was chromatographed on concanavalin A-Sepharose. The PGIP activity was bound by the lectin and then eluted using 250 mM alpha-methyl mannopyranoside, resulting in a 17-fold purification of the PGIP and demonstrating its glycoprotein nature. The anion-exchange and size-exclusion chromatography steps that followed gave a PGIP that was 857-fold purified relative to the initial tissue extract, and having a 44 kDa molecular weight, as estimated by SDS-PAGE electrophoresis. PGIP inhibition activity was tested with endo-polygalacturonase (EC 3.2.1.15) produced by Penicillium italicum and Botrytis cinerea. The radial diffusion and reducing sugar assays showed that P. italicum and B. cinerea endo-PGs were affected by PGIP, whereas no endo-PG activity was detected in the culture filtrate of P. digitatum. In vitro tests revealed that PGIP inhibited P. italicum and B. cinerea growth. By contrast, the influence of PGIP on P. digitatum, growth was negligible, perhaps because this fungus does not produce endo-PG. Following heating for 10 min at 65 degrees C the inhibitory activity of PGIP was reduced by 43%. PGIP activity decreased further as heating temperature increased, and was completely suppressed after heating at 100 degrees C for 10 min.
authors.affiliation: C.N.R. - Istituto di Scienze delle Produzioni Alimentari, Sezione di Sassari, Italy.
authors.forename: Guy
authors.initials: G
authors.lastname: D'hallewin
authors.forename: Mario
authors.initials: M
authors.lastname: Schirra
authors.forename: Ann L. T.
authors.initials: AL
authors.lastname: Powell
authors.forename: L. Carl
authors.initials: LC
authors.lastname: Greve
authors.forename: John M.
authors.initials: JM
authors.lastname: Labavitch