Analysis of the Specificity and Biochemical Characterization of Metalloproteases Isolated from Eupenicillium javanicum Using Fluorescence Resonance Energy Transfer Peptides

Analysis of the Specificity and Biochemical Characterization of Metalloproteases Isolated from Eupenicillium javanicum Using Fluorescence Resonance Energy Transfer Peptides

Author Hamin Neto, Youssef A. A. Google Scholar
de oliveira, Lilian C. G.[INIFESP] Google Scholar
de oliveira, Juliana R.[INIFESP] Google Scholar
Juliano, Maria A.[INIFESP] Google Scholar
Juliano, Luiz[INIFESP] Google Scholar
Arantes, Eliane C. Google Scholar
Cabral, Hamilton Google Scholar
Abstract Enzymes have important features that may facilitate their application in industrial processes and have been used as alternatives to chemical catalysts. In particular, proteases can be isolated from microorganisms, which provide important sources of advantageous enzymes for industrial processes. For example, Eupenicillium javanicum is a filamentous fungus that has been shown to express industrially applicable enzymes and chemical components, such as antifungal compounds. The biotechnological potential of E. javanicum and proteases made us search a novel protease from this microorganism. The macromolecule was isolated, the main biochemical properties was evaluated, and the specificity of the protease subsites was determined. The protease was produced under solid-state bioprocess with wheat bran and isolated by two chromatography steps with yield of 27.5% and 12.4-fold purification. The molecular mass was estimated at 30 kDa. The N-terminal sequence of the first 20 amino acid residues was AVGAGYNASVALALEKALNN. The enzyme presented higher proteolytic activity at pH 6.0 and 60 degrees C. The protease is stable at wide range of pH values and temperatures and in the presence of surfactants. The primed side of the catalytic site showed the highest catalytic efficiency of the enzyme isolated from E. javanicum. The S'(1) subsite is responsible for catalyzing the protease reaction with substrates with tyrosine in P'(1). These findings provide important insights into the biochemical characterization of a highly active protease from E. javanicum and may facilitate the development of industrial processes involving this protease.
Keywords biochemical characterization
fluorescence resonance energy transfer peptides
microbial enzyme
protease
solid-state fermentation
xmlui.dri2xhtml.METS-1.0.item-coverage Lausanne
Language English
Sponsor Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2012/24703-8, 2011/06986-0]
Grant number FAPESP: 2012/24703-8
FAPESP: 2011/06986-0
Date 2017
Published in Frontiers In Microbiology. Lausanne, v. 7, p. -, 2017.
ISSN 1664-302X (Sherpa/Romeo, impact factor)
Publisher Frontiers Media Sa
Extent -
Origin http://dx.doi.org/10.3389/fmicb.2016.02141
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000391497100001
URI https://repositorio.unifesp.br/handle/11600/55264

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