title: Biochemical study of the extracellular aspartyl protease Eap1 from the phytopathogen fungus Sporisorium reilianum.
abstract: In this work, the extracellular protease Eap1 from Sporisorium reilianum was characterized in solid and liquid cultures using different culture media. The results showed that Eap1 was produced in all media and under all culture conditions, with the most activity in solid culture at an acidic pH of 3-5. Following purification, the 41 kDa protease demonstrated aspartyl protease activity. The enzyme was stable at a wide range of temperatures and pH values, but 45°C and pH 3 were optimal. The K(m) and V(max( values obtained were 0.69 mg/mL and 0.66 μmol/min, respectively, with albumin as the substrate. Eap1 degraded hemoglobin as well as proteins obtained from corn germ, roots, stems and slides at pH 3 and also had milk-clotting activity. Sequencing analysis showed that this protein has 100% similarity to the peptide sequence theoretically obtained from the sr11394 gene, which encodes an aspartyl protease secreted by S. reilianum.
authors.affiliation: Universidad Politécnica de Pachuca, Carretera Pachuca-Cd. Sahagún, Km 20, Rancho Luna, Ex-Hacienda de Sta. Bárbara, Municipio de Zempoala, Hidalgo, Mexico.
authors.forename: Virginia
authors.initials: V
authors.lastname: Mandujano-González
authors.forename: Ainhoa
authors.initials: A
authors.lastname: Arana-Cuenca
authors.forename: Miguel Ángel
authors.initials: MÁ
authors.lastname: Anducho-Reyes
authors.forename: Alejandro
authors.initials: A
authors.lastname: Téllez-Jurado
authors.forename: Aldo E
authors.initials: AE
authors.lastname: González-Becerra
authors.forename: Yuridia
authors.initials: Y
authors.lastname: Mercado-Flores
mesh_terms: D000418:Albumins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D057055:Aspartic Acid Proteases / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism; D005110:Extracellular Space; D005656:Fungal Proteins / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D013696:Temperature; D014587:Ustilaginales / Q000737:chemistry / Q000201:enzymology*
chemical_list: D000418:Albumins; D003470:Culture Media; D005656:Fungal Proteins; D057055:Aspartic Acid Proteases
keywords: Aspartyl protease; Phytopathogenic fungus; Sporisorium reilianum