Characterization of arazyme, an exocellular metalloprotease isolated from Serratia proteamaculans culture medium

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dc.contributor.author Bersanetti, P. A.
dc.contributor.author Park, H. Y.
dc.contributor.author Bae, K. S.
dc.contributor.author Son, K. H.
dc.contributor.author Shin, D. H.
dc.contributor.author Hirata, I. Y.
dc.contributor.author Juliano, M. A.
dc.contributor.author Carmona, A. K.
dc.contributor.author Juliano, L.
dc.date.accessioned 2016-01-24T12:38:08Z
dc.date.available 2016-01-24T12:38:08Z
dc.date.issued 2005-11-01
dc.identifier http://dx.doi.org/10.1016/j.enzmictec.2005.01.041
dc.identifier.citation Enzyme and Microbial Technology. New York: Elsevier B.V., v. 37, n. 6, p. 574-581, 2005.
dc.identifier.issn 0141-0229
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/28527
dc.description.abstract We investigated the biochemical properties of a 51.5 kDa metalloprotease (arazyme, 3.4.24.40) secreted into the culture medium by Aranicola proteolyticus, a symbiotic bacterium of the spider Nephila clavata. the enzyme was purified to apparent homogeneity by ion exchange chromatography in a Resource Q column (FPLC system). the substrate specificity requirements of purified arazyme were examined using fluorescence resonance energy transfer (FRET) peptides derived from the lead sequence Abz-KLRFSKQ-EDDnp (Abz = ortho-aminobenzoic acid; EDDnp=ethylenediaminedinitrophenyl). Three series of peptides were assayed to map the S-2, S-1 and S'(1) subsites: Abz-KXRFSKQ-EDDnp, Abz-KLXFSKQ-EDDnp and Abz-KLRXSKQ-EDDnp (X are natural amino acids). the results indicated that S-1 subsite has a broad specificity, being Gly the preferred amino acid for this subsite followed by positively charged residues (Arg and His). the S-2 and S'(1) subsites accommodated better hydrophobic residues with aliphatic or aromatic side chains (Leu, Phe). the pH effect on hydrolysis of Abz-KLFFSKQ-EDDnp indicated that optimal hydrolysis occurred at pH 8.0 or higher. the effect of NaCl on the arazyme activity depends on the substrate, but in general the activity was reduced with this salt. the temperature did not affect the enzyme from 10 to 45 degrees C, after which activity decreased sharply. Arazyme presented high hydrolytic activity on substance P and peptides related to bradykinin. in addition, arazyme activity was resistant to the treatment by pepsin, trypsin and chymotrypsin. in conclusion, arazyme has a broad hydrolytic profile and works in very aggressive conditions, which justify its potential use in therapeutics and biotechnological applications. (c) 2005 Elsevier Inc. All rights reserved. en
dc.format.extent 574-581
dc.language.iso eng
dc.publisher Elsevier B.V.
dc.relation.ispartof Enzyme and Microbial Technology
dc.rights Acesso restrito
dc.subject serralysin en
dc.subject Serratia proteamaculans en
dc.subject fluorescent peptides en
dc.title Characterization of arazyme, an exocellular metalloprotease isolated from Serratia proteamaculans culture medium en
dc.type Artigo
dc.rights.license http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.contributor.institution Korea Res Inst Biosci & Biotechnol
dc.contributor.institution Insect Biotech Co Ltd
dc.description.affiliation UNIFESP, Dept Biophys, Escola Paulista Med, BR-04044020 São Paulo, Brazil
dc.description.affiliation Korea Res Inst Biosci & Biotechnol, Insect Resources Lab, Taejon 305333, South Korea
dc.description.affiliation Insect Biotech Co Ltd, Taejon 305811, South Korea
dc.description.affiliationUnifesp UNIFESP, Dept Biophys, Escola Paulista Med, BR-04044020 São Paulo, Brazil
dc.identifier.doi 10.1016/j.enzmictec.2005.01.041
dc.description.source Web of Science
dc.identifier.wos WOS:000232363400002



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