TY - JOUR
T1 - Biochemical and thermodynamic characteristics of a new serine protease from Mucor subtilissimus URM 4133
AU - Gomes, José Erick Galindo
AU - Rosa, Isabel Zaparoli
AU - Nascimento, Talita Camila Evaristo da Silva
AU - Souza-Motta, Cristina Maria de
AU - Gomes, Eleni
AU - Boscolo, Mauricio
AU - Moreira, Keila Aparecida
AU - Pintado, Maria Manuela Estevez
AU - Silva, Roberto da
N1 - Funding Information:
This work received financial support from the São Paulo Research Foundation (FAPESP) (grant numbers 2014/13700-3 and 2017/16482-5 ) and the National Council for Scientific and Technological Development (CNPQ) [grant number 426578/2016-3 ]. We would also like to thank the scientific collaboration of CBQF under the FCT project UID/Multi/50016/2013.
Funding Information:
This work received financial support from the São Paulo Research Foundation (FAPESP) (grant numbers 2014/13700-3 and 2017/16482-5) and the National Council for Scientific and Technological Development (CNPQ) [grant number 426578/2016-3]. We would also like to thank the scientific collaboration of CBQF under the FCT project UID/Multi/50016/2013.
Publisher Copyright:
© 2020 The Authors
PY - 2020/12
Y1 - 2020/12
N2 - 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.
AB - 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.
KW - Enzymatic characterization
KW - Mucor subtilissimus
KW - Peptide sequences by MALDI-TOF
KW - Serine protease
UR - http://www.scopus.com/inward/record.url?scp=85096704864&partnerID=8YFLogxK
U2 - 10.1016/j.btre.2020.e00552
DO - 10.1016/j.btre.2020.e00552
M3 - Article
C2 - 33294402
AN - SCOPUS:85096704864
SN - 2215-017X
VL - 28
JO - Biotechnology Reports
JF - Biotechnology Reports
M1 - e00552
ER -