TY - JOUR
T1 - Intracellular Aminopeptidase Activity Determination from the Fungus Sporisorium reilianum
T2 - Purification and Biochemical Characterization of psrAPEi Enzyme
AU - Pérez-Rodríguez, Joany
AU - Téllez-Jurado, Alejandro
AU - Villa-Tanaca, Lourdes
AU - Gómez-Aldapa, Carlos Alberto
AU - Mercado-Flores, Yuridia
N1 - Publisher Copyright:
© 2022, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2022/3
Y1 - 2022/3
N2 - The aims of this study were to, first, determine the intracellular aminopeptidase activity (APEi) and second, purify and biochemically characterize one intracellular aminopeptidase enzyme from the phytopathogen fungus Sporisorium reilianum (psrAPEi), the causal agent of head smut in corn. The fungus produced APEi activity in all media cultures evaluated. The psrAPEi was purified by a procedure that involved ammonium sulfate fractionation and four chromatographic steps using an FPLC system (Fast Protein Liquid Chromatography). Results showed an estimated molecular mass of 52.2 kDa. Enzymatic activity was optimal at pH 7.0 and 35 °C and was inhibited by EDTA-Na2, 1,10-phenanthroline, bestatin, and PMSF. This aminopeptidase showed a preference for leucine, arginine, and lysine at the N-position. The Km and Vmax values were 3.72 μM and 188.0 μmol/min, respectively, for l-lysyl-4-nitroanilide. This is the first study to report on intracellular aminopeptidase activity in S. reilianum and the purification and characterization of an intracellular metallo-serine-aminopeptidase (psrAPEi).
AB - The aims of this study were to, first, determine the intracellular aminopeptidase activity (APEi) and second, purify and biochemically characterize one intracellular aminopeptidase enzyme from the phytopathogen fungus Sporisorium reilianum (psrAPEi), the causal agent of head smut in corn. The fungus produced APEi activity in all media cultures evaluated. The psrAPEi was purified by a procedure that involved ammonium sulfate fractionation and four chromatographic steps using an FPLC system (Fast Protein Liquid Chromatography). Results showed an estimated molecular mass of 52.2 kDa. Enzymatic activity was optimal at pH 7.0 and 35 °C and was inhibited by EDTA-Na2, 1,10-phenanthroline, bestatin, and PMSF. This aminopeptidase showed a preference for leucine, arginine, and lysine at the N-position. The Km and Vmax values were 3.72 μM and 188.0 μmol/min, respectively, for l-lysyl-4-nitroanilide. This is the first study to report on intracellular aminopeptidase activity in S. reilianum and the purification and characterization of an intracellular metallo-serine-aminopeptidase (psrAPEi).
UR - http://www.scopus.com/inward/record.url?scp=85124756025&partnerID=8YFLogxK
U2 - 10.1007/s00284-022-02787-8
DO - 10.1007/s00284-022-02787-8
M3 - Artículo
C2 - 35129692
AN - SCOPUS:85124756025
SN - 0343-8651
VL - 79
JO - Current Microbiology
JF - Current Microbiology
IS - 3
M1 - 90
ER -