Advanced search
Start date
Betweenand

Production and characterization of chimeric proteins containing chitinase catalytic domain and modules for chitin binding

Abstract

Chitin, a polymer formed by monomers of ²-1,4-N-acetylglucosamine and second most biopolymer occurring on the planet after cellulose, is degraded by enzymes called chitinases (EC 3.2.2.14) that cleave the chitin chain at sites in a random manner. These enzymes are produced by a wide variety of organisms including bacteria, fungi, insects, plants and animals and have different functions such as nutrition, morphogenesis and defense against pathogens. In addition to the important biological function, chitinases have been used in biocontrol (activities as fungicidal, bactericide, insecticide). In insects chitinases have been related to the chitin remodeling, present in the exoskeleton and in the peritrophic matrix. Insect chitinases have been classified into eight distinct groups based primarily on the architecture of domains such as catalytic and CBM ('carbohydrate-binding module'). As a result of studies conducted in our research group on chitin synthesis and considering the lack of information on chitinase ants, we propose, in this project, the production of different chimeric proteins (containing chitinase catalytic domain and modules for binding in chitin) and the study of its catalytic function against chitin and analysis of the products through HPLC and mass spectrometry. Analysis of sequences deposited in databases has already been started and two pairs of oligonucleotides have been acquired. RNA from A. sexdens will be used to obtain cDNA that will be used as a template molecule in PCR for DNA amplification. The proteins expression of will be done in P. pastoris, a well established system in our Laboratory. It is also the objective of the project to evaluate if the proteins present potential use in biocontrol. To develop these experiments the proteins will be tested against the fungi A. fumigatus and C. albicans, important infectious agents in humans and against Spodoptera frugiperda, the main pest of the corn crop in Brazil. To better understand the protein-substrate interaction studies will be performed using the Nuclear Magnetic Resonance (NMR) technique. With this project, an effective contribution to the construction of protein modules with interesting activities, such as insecticide and fungicide, is expected. (AU)

Articles published in Agência FAPESP Newsletter about the research grant:
More itemsLess items
Articles published in other media outlets ( ):
More itemsLess items
VEICULO: TITULO (DATA)
VEICULO: TITULO (DATA)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DIONIZIO, BRUNA SOARES; SILVA RABELO, CAMILA ABREU B.; RAMOS DE JESUS, HUGO CESAR; AMANCIO VARESCHE, MARIA BERNADETE; FERREIRA DE SOUZA, DULCE HELENA. The Deconstruction of the Lignocellulolytic Structure of Sugarcane Bagasse by Laccases Improves the Production of H-2 and Organic Acids. Applied Biochemistry and Biotechnology, v. 194, n. 7, p. 22-pg., . (18/26470-7, 17/15455-4, 18/06297-9)
SANTOS CORREA, KATIA CELINA; MOREIRA, ARIELE CRISTINA; IBRAHIM, AMR GALAL ABD EL-RAHEEM; RAMOS DE JESUS, HUGO CESAR; MICOCCI, KELLI CRISTINA; CRIZOSTOMO KOCK, FLAVIO VINICIUS; BUENO, ODAIR C.; VENANCIO, TIAGO; HENRIQUE-SILVA, FLAVIO; SOUZA, DULCE HELENA F.. Identification and characterization of a recombinant cysteine peptidase (AsCathL) from leaf-cutting ant Atta sexdens Linnaeus, 1758 (Hymenoptera, Formicidae). Protein Expression and Purification, v. 201, p. 10-pg., . (17/15455-4, 15/21517-7, 14/12169-2, 18/16040-5, 18/06297-9)
MICOCCI, KELLI C.; MOREIRA, ARIELE C.; SANCHEZ, AMANDA D.; PETTINATTI, JESSICA L.; ROCHA, MARINA C.; DIONIZIO, BRUNA S.; CORREA, KATIA C. S.; MALAVAZI, IRAN; WOUTERS, FELIPE C.; BUENO, ODAIR C.; et al. Identification, cloning, and characterization of a novel chitinase from leaf-cutting ant Atta sexdens: An enzyme with antifungal and insecticidal activity. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1867, n. 1, p. 10-pg., . (12/25299-6, 17/06198-8, 17/19694-3, 18/06297-9)

Please report errors in scientific publications list using this form.
X

Report errors in this page


Error details: