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Type: Artigo de periódico
Title: Purification, characterization and structural determination of chitinases produced by Moniliophthora perniciosa
Author: Galante, Rafaela S.
Taranto, Alex G.
Koblitz, Maria G. B.
Goes-Neto, Aristoteles
Pirovani, Carlos P.
Cascardo, Julio C. M.
Cruz, Sandra H.
Pereira, Goncalo A. G.
de Assisi, Sandra A.
Abstract: The enzyme chitinase from Moniliophthora perniciosa the causative agent of the witches' broom disease in Theobroma cacao, was partially purified with ammonium sulfate and filtration by Sephacryl S-200 using sodium phosphate as an extraction buffer. Response surface methodology (RSM) was used to determine the optimum pH and temperature conditions. Four different isoenzymes were obtained: ChitMp I, ChitMp II, ChitMp III and ChitMp IV. ChitMp I had an optimum temperature at 44-73 degrees C and an optimum pH at 7.0-8.4. ChitMp II had an optimum temperature at 45-73 degrees C and an optimum pH at 7.0-8.4. ChitMp III had an optimum temperature at 54-67 degrees C and an optimum pH at 7.3-8.8. ChitMp IV had an optimum temperature at 60 degrees C and an optimum pH at 7.0. For the computational biology, the primary sequence was determined in silico from the database of the Genome/Proteome Project of M. perniciosa, yielding a sequence with 564 bp and I 88 amino acids that was used for the three-dimensional design in a comparative modeling methodology. The generated models were submitted to validation using Procheck 3.0 and ANOLEA. The model proposed for the chitinase was subjected to a dynamic analysis over a 1 ns interval, resulting in a model with 91.7% of the residues occupying favorable places on the Ramachandran plot and an RMS of 2.68.
Subject: Chitinase
Moniliophthora perniciosa
kinetic characterization
heat stability
3D structure
comparative modeling
Country: Brazil
Editor: Acad Brasileira De Ciencias
Citation: Anais da Academia Brasileira de Ciencias. Acad Brasileira De Ciencias, v.84, n.2, p.469-486, 2012
Rights: fechado
Date Issue: 2012
Appears in Collections:IB - Artigos e Outros Documentos

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