Merheb-Dini, Carolina [UNESP]Cabral, Hamilton [UNESP]Leite, Rodrigo S. R. [UNESP]Zanphorlin, Leticia M. [UNESP]Okamoto, Debora N. [UNESP]Bonilla-Rodriguez, Gustavo Orlando [UNESP]Juliano, LuizArantes, Eliane C.Gomes, Eleni [UNESP]da Silva, Roberto [UNESP]2014-05-202014-05-202009-10-14Journal of Agricultural and Food Chemistry. Washington: Amer Chemical Soc, v. 57, n. 19, p. 9210-9217, 2009.0021-8561http://hdl.handle.net/11449/21609Protease production was carried out in solid state fermentation. The enzyme was purified through precipitation with ethanol at 72% followed by chromatographies in columns of Sephadex G75 and Sephacryl S100. It was purified 80-fold and exhibited recovery of total activity of 0.4%. SDS-PAGE analysis indicated an estimated molecular mass of 24.5 kDa and the N-terminal sequence of the first 22 residues was APYSGYQCSMQLCLTCALMNCA. Purified protease was only inhibited by EDTA (96.7%) and stimulated by Fe(2+) revealing to be a metalloprotease activated by iron. Optimum pH was 5.5, optimum temperature was 75 degrees C, and it was thermostable at 65 degrees C for 1 h maintaining more than 70% of original activity. Through enzyme kinetic studies, protease better hydrolyzed casein than azocasein. The screening of fluorescence resonance energy transfer (FRET) peptide series derived from Abz-KLXSSKQ-EDDnp revealed that the enzyme exhibited preference for Arg in P(1) (k(cat)/K(m) = 30.1 mM(-1) s(-1)).9210-9217engThermoascus aurantiacusSolid state fermentation (SSF)metalloproteaseThermostabilityfluorescent peptidesN-terminal sequenceArtigo10.1021/jf9017977WOS:000270461500067Acesso restrito695525858867213070912417428519209424175688206545