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Literature summary extracted from

  • Benedetti, M.; Andreani, F.; Leggio, C.; Galantini, L.; Di Matteo, A.; Pavel, N.V.; De Lorenzo, G.; Cervone, F.; Federici, L.; Sicilia, F.
    A single amino-acid substitution allows endo-polygalacturonase of Fusarium verticillioides to acquire recognition by PGIP2 from Phaseolus vulgaris (2013), PLoS ONE, 8, e80610.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.15 cloning in Escherichia coli strain TOP10F, expression in Pichia pastoris strain X-33 and secretion to the culture medium Fusarium verticillioides
3.2.1.15 recombinant enzyme expression Fusarium phyllophilum

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.15 A274T site-directed mutagenesis, the mutation causes a marked loss of inhibition by PvPGIP2 of 150fold Fusarium phyllophilum
3.2.1.15 K116E site-directed mutagenesis, the mutant shows no inhibition by PvPGIP2 Fusarium phyllophilum
3.2.1.15 K310T site-directed mutagenesis Fusarium phyllophilum
3.2.1.15 L303E site-directed mutagenesis Fusarium verticillioides
3.2.1.15 L303E site-directed mutagenesis, the mutant enzyme is inhibited by Inhibitor mutant PvPGIP2.Q224K in contrast to the wild-type enzyme Fusarium phyllophilum
3.2.1.15 N121K site-directed mutagenesis, the mutation only slightly affects inhibition by PvPGIP2 Fusarium phyllophilum
3.2.1.15 Q124P site-directed mutagenesis, the mutation only slightly affects inhibition by PvPGIP2 Fusarium phyllophilum
3.2.1.15 S120N site-directed mutagenesis, the mutation only slightly affects inhibition by PvPGIP2 Fusarium phyllophilum
3.2.1.15 S120N/N121K/S122D site-directed mutagenesis, the triple mutant is inhibited with a 25fold reduced efficiency by PvPGIP2 Fusarium phyllophilum
3.2.1.15 S122D, site-directed mutagenesis, the mutation only slightly affects inhibition by PvPGIP2 Fusarium phyllophilum
3.2.1.15 S363K site-directed mutagenesis Fusarium phyllophilum
3.2.1.15 T274A site-directed mutagenesis Fusarium verticillioides

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.15 additional information inability of the enzyme from Fusarium phyllophilum to be inhibited by polygalacturonase-inhibiting protein 2, PGIP2, from Phaseolus vulgaris pv. Pinto and recombinatly expressed in Pichia pastoris X33, or any other polygalacturonase-inhibiting protein, small angle X-ray scattering interaction analysis of structural basis, overview Fusarium verticillioides
3.2.1.15 polygalacturonase-inhibiting protein 2 PGIP2, from Phaseolus vulgaris pv. Pinto, recombinantly expressed in Pichia pastoris X33, forms a complex with the enzyme from Colletotrichum lupini hindering substrate binding in the enzyme active site cleft, small angle X-ray scattering interaction analysis, and inhibition kinetics, overview Colletotrichum lupini
3.2.1.15 polygalacturonase-inhibiting protein 2 PGIP2, from Phaseolus vulgaris pv. Pinto, recombinantly expressed in Pichia pastoris X33, forms a complex with the enzyme from Fusarium phyllophilum hindering substrate binding in the enzyme active site cleft, residue A274 plays a key role in the interaction, small angle X-ray scattering interaction analysis, and inhibition kinetics, overview. Residue Q224 of the inhibitor is crucial for the interaction with the enzyme. Inhibitor mutant PvPGIP2.Q224K variant is unable to inhibit the enzyme, while inhibitor mutant PvPGIP2.Q224K retains unaltered inhibitory capability towards the enzyme Fusarium phyllophilum

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.15 extracellular
-
Colletotrichum lupini
-
-

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.15 Colletotrichum lupini
-
-
-
3.2.1.15 Colletotrichum lupini SHK78813
-
-
-
3.2.1.15 Fusarium phyllophilum
-
-
-
3.2.1.15 Fusarium verticillioides
-
-
-
3.2.1.15 Fusarium verticillioides 62264
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.15 native extracellular enzyme from culture medium Colletotrichum lupini
3.2.1.15 recombinant enzyme Fusarium phyllophilum
3.2.1.15 recombinant extracellular enzyme from Pichia pastoris strain X-33 culture medium by ultrafiltration and dialysis, anion and cation exchange chromatography, dialysis, and hydrophobic interaction chromatography Fusarium verticillioides

Synonyms

EC Number Synonyms Comment Organism
3.2.1.15 CluPG1
-
Colletotrichum lupini
3.2.1.15 FgPG
-
Fusarium verticillioides
3.2.1.15 FpPG
-
Fusarium phyllophilum