Autor(es):
Gomes, José Erick Galindo [UNESP] ; Rosa, Isabel Zaparoli [UNESP] ; Nascimento, Talita Camila Evaristo da Silva ; Souza-Motta, Cristina Maria de ; Gomes, Eleni [UNESP] ; Boscolo, Mauricio [UNESP] ; Moreira, Keila Aparecida ; Pintado, Maria Manuela Estevez ; da Silva, Roberto [UNESP]
Data: 2021
Identificador Persistente: http://hdl.handle.net/11449/208186
Origem: Oasisbr
Assunto(s): Enzymatic characterization; Mucor subtilissimus; Peptide sequences by MALDI-TOF; Serine protease; Enzymatic characterization; Enzymatic characterization; Mucor subtilissimus; Mucor subtilissimus; Peptide sequences by MALDI-TOF; Peptide sequences by MALDI-TOF; Serine protease; Serine protease
Descrição
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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.
São Paulo State University (UNESP) Institute of Biosciences Humanities and Exact Sciences (IBILCE) Cristovão Colombo, 2265, Jardim Nazareth
Laboratory of Microbiology Enzymatic Technology and Bioproducts Academic Unit of Garanhuns Federal Rural University of Pernambuco (UFRPE), Bom Pastor Avenue
Department of Mycology Center of Biosciences Federal University of Pernambuco (UFPE), Prof. Nelson Chaves Avenue
Centro de Biotecnologia e Química Fina Universidade Católica Portuguesa (UCP) – Escola Superior de Biotecnologia, Rua Arquiteto Lobão Vital, 172
São Paulo State University (UNESP) Institute of Biosciences Humanities and Exact Sciences (IBILCE) Cristovão Colombo, 2265, Jardim Nazareth
FAPESP: 2014/13700-3
FAPESP: 2017/16482-5
CNPq: 426578/2016-3
