title: Production of an extracellular trypsin-like protease by the fungal plant pathogen Verticillium dahliae.
abstract: The plant pathogenic fungus Verticillium dahliae produced extracellular alkaline protease activity when grown in liquid medium supplemented with a protein source. A serine protease was purified 80-fold in a single step, using cation-exchange chromatography, from the filtrate of cultures grown with skim milk as a protein source. N-terminal amino acid sequence analysis of the 30-kDa protein (VDP30) that copurified with the serine protease activity suggested that VDP30 is a trypsin-like protein. The purified enzyme hydrolyzed the synthetic substrate N alpha-benzoyl-DL-arginine p-nitroanilide hydrochloride (BAPNA), and the activity on BAPNA was inhibited by leupeptin, further verifying the trypsin-like nature of the enzyme.
authors.affiliation: Pest Management Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada. dobinsonk@em.agr.ca
authors.forename: K F
authors.initials: KF
authors.lastname: Dobinson
authors.forename: N
authors.initials: N
authors.lastname: Lecomte
authors.forename: G
authors.initials: G
authors.lastname: Lazarovits
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001586:Benzoylarginine Nitroanilide / Q000378:metabolism; D018533:Biomass; D002852:Chromatography, Ion Exchange; D003470:Culture Media / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007976:Leupeptins / Q000494:pharmacology; D008892:Milk / Q000378:metabolism; D003904:Mitosporic Fungi / Q000201:enzymology* / Q000378:metabolism*; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013997:Time Factors
chemical_list: D003470:Culture Media; D007976:Leupeptins; D015842:Serine Proteinase Inhibitors; D001586:Benzoylarginine Nitroanilide; D012697:Serine Endopeptidases; C032854:leupeptin