Determination of Specificity and Biochemical Characteristics of Neutral Protease Isolated from Myceliophthora thermophila

Author(s)

Hamin Neto, Youssef A. A. ^[1] ; de Oliveira, Lilian C. G. ^[2] ; de Oliveira, Arthur H. C. ^[3] ; Rosa, Jose C. ^[4] ; Juliano, Maria A. ^[2] ; Juliano, Luiz ^[2] ; Rodrigues, Andre ^[5] ; Cabral, Hamilton ^[1]

Affiliation

^[1] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Dept Pharmaceut Sci, BR-14049 Ribeirao Preto - Brazil

^[2] Univ Fed Sao Paulo, Dept Biophys, Sao Paulo - Brazil

^[3] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Chem, BR-14049 Ribeirao Preto - Brazil

^[4] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Mol Cell Biol & Pathogen Bioagents, BR-14049 Ribeirao Preto - Brazil

^[5] Univ Estadual Paulista, Dept Biochem & Microbiol, Rio Claro - Brazil

• Document type: Journal article
• Source: PROTEIN AND PEPTIDE LETTERS; v. 22, n. 11, p. 972-982, 2015.
• Web of Science Citations: 6

Abstract

Proteases hydrolyze polypeptides to release peptides and/or amino acids. This subclass of enzymes is among those with the most sales worldwide, particularly those produced by microorganisms. Proteases may be applied in the several industries, including the food industry, leather, detergents, and bioremediation. Myceliophthora thermophila protease was produced by a submerged bioprocess and then purified 185-fold by anion exchange and hydrophobic chromatography with a 37% yield. The molecular mass was estimated at 36.2 kDa, and mass spectrometry identified two sequences: GVVANMSLGGSYSASINNAAAALVR and STGNAAITGVPSGTTNR. The isolated protein was characterized biochemically, showed an optimum pH of 6.5 and optimum temperature of 45 degrees C, and stability at wide range of pH and temperatures and in the presence of reducing agents and some surfactants. Kinetic assays for this enzyme showed a greater catalytic efficiency when the substrate had alanine at position P-2'. The protease presented characteristics that may be of interest to many industrial areas. (AU)

• FAPESP's process: 12/50191-4 - Synthesis, kinetic studies and applications of substrates and inhibitors for proteolytic enzymes
• Grantee: Maria Aparecida Juliano
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 12/18278-2 - Bioprocess, purification and characterization of peptidases produced by fungi Eupenicillium javanicum and Myceliophthora sp. and evaluating the production of biologically active peptides
• Grantee: Youssef Ali Abou Hamin Neto
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 11/06986-0 - Determination of the specificity of peptidases isolated from fungi using fluorescence resonance energy transfer (FRET) peptides as substrates
• Grantee: Hamilton Cabral
• Support Opportunities: Regular Research Grants
• FAPESP's process: 12/24703-8 - Proteomics analyses of meso and thermophilic filamentous fungi exposed to physical and chemical factors
• Grantee: Hamilton Cabral
• Support Opportunities: Regular Research Grants