Biochemical and thermodynamic characteristics of a new serine protease from Mucor subtilissimus URM 4133

José Erick Galindo Gomes, Isabel Zaparoli Rosa, Talita Camila Evaristo da Silva Nascimento, Cristina Maria de Souza-Motta, Eleni Gomes, Mauricio Boscolo, Keila Aparecida Moreira, Maria Manuela Estevez Pintado, Roberto da Silva*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

A protease from the fungus Mucor subtilissimus URM 4133, capable of producing bioactive peptides from goat casein, was purified. SDS-PAGE and zymography showed a molecular mass of 30 kDa. The enzyme was active and stable in a wide pH range (6.0–10.5) and (5.0–10.5), respectively. Optimum temperature was at 45–50 °C and stability was above 80 % (40 °C/2 h). Activity was not influenced by ions or organic substances (Triton, Tween, SDS and DMSO), but was completely inhibited by PMSF, suggesting that it belongs to the serine protease family. The Km and Vmax were 2.35 mg azocasein.mL-1 and 333.33 U.mg protein-1, respectively. Thermodynamic parameters of irreversible denaturation (40–60 °C) were enthalpy 123.63 – 123.46 kJ.mol-1, entropy 120.24–122.28 kJ.mol-1 and Gibbs free energy 85.97 – 82.45 kJ.mol-1. Any peptide sequences compatible with this protease were found after analysis by MALDI-TOF, which suggests that it is a new serine protease.

Original languageEnglish
Article numbere00552
JournalBiotechnology Reports
Volume28
DOIs
Publication statusPublished - Dec 2020

Keywords

  • Enzymatic characterization
  • Mucor subtilissimus
  • Peptide sequences by MALDI-TOF
  • Serine protease

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