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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Merheb-Dini, Carolina [1] ; Cabral, Hamilton [2] ; Leite, Rodrigo S. R. [1] ; Zanphorlin, Leticia M. [1] ; Okamoto, Debora N. [1] ; Bonilla Rodriguez, Gustavo O. [1] ; Juliano, Luiz [3] ; Arantes, Eliane C. [2] ; Gomes, Eleni [1] ; da Silva, Roberto [1]
Total Authors: 10
[1] Univ Estadual Paulista, Lab Bioquim & Microbiol Aplicada, Inst Biociencias Letras & Ciencias Exatas, BR-15054000 Sao Paulo - Brazil
[2] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, BR-14040903 Sao Paulo - Brazil
[3] Univ Fed Sao Paulo, Dept Biofis, Escola Paulista Med, BR-04044020 Sao Paulo - Brazil
Total Affiliations: 3
Document type: Journal article
Source: Journal of Agricultural and Food Chemistry; v. 57, n. 19, p. 9210-9217, OCT 14 2009.
Web of Science Citations: 18

Protease 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)). (AU)

FAPESP's process: 06/06573-9 - Production, purification and characterization of Rhizomucor sp. and evaluation of its application in mature cheese production
Grantee:Carolina Merheb Dini
Support Opportunities: Scholarships in Brazil - Doctorate