Biochemical and Functional Characterization of a Metalloprotease from the Thermophilic Fungus Thermoascus aurantiacus

dc.contributorUniversidade Estadual Paulista (UNESP)
dc.creatorMerheb-Dini, Carolina
dc.creatorCabral, Hamilton
dc.creatorLeite, Rodrigo S. R.
dc.creatorZanphorlin, Leticia M.
dc.creatorOkamoto, Debora N.
dc.creatorBonilla-Rodriguez, Gustavo Orlando
dc.creatorJuliano, Luiz
dc.creatorArantes, Eliane C.
dc.creatorGomes, Eleni
dc.creatorda Silva, Roberto
dc.date2014-05-20T14:01:08Z
dc.date2016-10-25T17:08:27Z
dc.date2014-05-20T14:01:08Z
dc.date2016-10-25T17:08:27Z
dc.date2009-10-14
dc.date.accessioned2017-04-05T21:24:52Z
dc.date.available2017-04-05T21:24:52Z
dc.identifierJournal of Agricultural and Food Chemistry. Washington: Amer Chemical Soc, v. 57, n. 19, p. 9210-9217, 2009.
dc.identifier0021-8561
dc.identifierhttp://hdl.handle.net/11449/21609
dc.identifierhttp://acervodigital.unesp.br/handle/11449/21609
dc.identifier10.1021/jf9017977
dc.identifierWOS:000270461500067
dc.identifierhttp://dx.doi.org/10.1021/jf9017977
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/867161
dc.descriptionProtease 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)).
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.languageeng
dc.publisherAmer Chemical Soc
dc.relationJournal of Agricultural and Food Chemistry
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectThermoascus aurantiacus
dc.subjectSolid state fermentation (SSF)
dc.subjectmetalloprotease
dc.subjectThermostability
dc.subjectfluorescent peptides
dc.subjectN-terminal sequence
dc.titleBiochemical and Functional Characterization of a Metalloprotease from the Thermophilic Fungus Thermoascus aurantiacus
dc.typeOtro


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