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Intracellular Aminopeptidase Activity Determination from the Fungus Sporisorium reilianum: Purification and Biochemical Characterization of psrAPEi Enzyme

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Abstract

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).

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Acknowledgements

We thank to the CONACyT for the scholarship given to Joany Pérez-Rodríguez.

Funding

This work was supported by the Fondo de Ciencia Básica SEP-CONACyT (Project No. 167459).

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Authors and Affiliations

  1. Universidad Politécnica de Pachuca, Zempoala, Hidalgo, Mexico

    Joany Pérez-Rodríguez, Alejandro Téllez-Jurado & Yuridia Mercado-Flores

  2. Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, Mexico

    Lourdes Villa-Tanaca

  3. Área Académica ICBI, Universidad Autónoma del Estado de Hidalgo, Mineral de la Reforma, Hidalgo, Mexico

    Carlos Alberto Gómez-Aldapa

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  1. Joany Pérez-Rodríguez
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  5. Yuridia Mercado-Flores
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Contributions

JPR performed the experiments and the analysis of the data. ATJ assisted in methodology and helped in the analysis of the data. LVT assisted in methodology and helped in the analysis of the data. CAGA helped to determine the thermal properties of the purified enzyme. YMF performed investigation, analysis of the data, project administration, and writing and reviewing of the manuscript. All authors have read the manuscript and their contributions are equal.

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Correspondence to Yuridia Mercado-Flores.

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In this work neither animal was used to the experimental development. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Pérez-Rodríguez, J., Téllez-Jurado, A., Villa-Tanaca, L. et al. Intracellular Aminopeptidase Activity Determination from the Fungus Sporisorium reilianum: Purification and Biochemical Characterization of psrAPEi Enzyme. Curr Microbiol 79, 90 (2022). https://doi.org/10.1007/s00284-022-02787-8

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