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Functional characterization and target discovery of glycoside hydrolases from the digestome of the lower termite Coptotermes gestroi

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Franco Cairo, Joao Paulo L. [1, 2] ; Leonardo, Flavia C. [3, 1] ; Alvarez, Thabata M. [2] ; Ribeiro, Daniela A. [2] ; Buechli, Fernanda [2] ; Costa-Leonardo, Ana M. [4] ; Carazzolle, Marcelo F. [5, 1] ; Costa, Fernando F. [3] ; Paes Leme, Adriana F. [5] ; Pereira, Goncalo A. G. [5, 1] ; Squina, Fabio M. [2]
Número total de Autores: 11
Afiliação do(s) autor(es):
[1] Univ Estadual Campinas, Genet Evolut & Bioagents Dept, LGE, Campinas, SP - Brazil
[2] CNPEM, Lab Nacl Cencia & Tecnol Bioetanol CTBE, Campinas, SP - Brazil
[3] Univ Estadual Campinas, Hematol & Hemotherapy Ctr, Campinas, SP - Brazil
[4] Univ Estadual Paulista UNESP, Inst Biocencias, Dept Biol, Rio Claro - Brazil
[5] CNPEM, Lab Nacl Biociencias LNBio, Campinas, SP - Brazil
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Citações Web of Science: 23
Assunto(s):Celulase   beta-Glucosidase

Background: Lignocellulosic materials have been moved towards the forefront of the biofuel industry as a sustainable resource. However, saccharification and the production of bioproducts derived from plant cell wall biomass are complex and lengthy processes. The understanding of termite gut biology and feeding strategies may improve the current state of biomass conversion technology and bioproduct production. Results: The study herein shows comprehensive functional characterization of crude body extracts from Coptotermes gestroi along with global proteomic analysis of the termite's digestome, targeting the identification of glycoside hydrolases and accessory proteins responsible for plant biomass conversion. The crude protein extract from C. gestroi was enzymatically efficient over a broad pH range on a series of natural polysaccharides, formed by glucose-, xylose-, mannan- and/or arabinose-containing polymers, linked by various types of glycosidic bonds, as well as ramification types. Our proteomic approach successfully identified a large number of relevant polypeptides in the C. gestroi digestome. A total of 55 different proteins were identified and classified into 29 CAZy families. Based on the total number of peptides identified, the majority of components found in the C. gestroi digestome were cellulose-degrading enzymes. Xylanolytic enzymes, mannan-hydrolytic enzymes, pectinases and starch-degrading and debranching enzymes were also identified. Our strategy enabled validation of liquid chromatography with tandem mass spectrometry recognized proteins, by enzymatic functional assays and by following the degradation products of specific 8-amino-1,3,6-pyrenetrisulfonic acid labeled oligosaccharides through capillary zone electrophoresis. Conclusions: Here we describe the first global study on the enzymatic repertoire involved in plant polysaccharide degradation by the lower termite C. gestroi. The biochemical characterization of whole body termite extracts evidenced their ability to cleave all types of glycosidic bonds present in plant polysaccharides. The comprehensive proteomic analysis, revealed a complete collection of hydrolytic enzymes including cellulases (GH1, GH3, GH5, GH7, GH9 and CBM 6), hemicellulases (GH2, GH10, GH11, GH16, GH43 and CBM 27) and pectinases (GH28 and GH29). (AU)

Processo FAPESP: 06/59086-8 - Estudo do cupim coptotermes gestroi: analise da base genetica da diferenciacao de castas e busca de genes de importancia biotecnologica
Beneficiário:Flávia Costa Leonardo
Linha de fomento: Bolsas no Brasil - Doutorado
Processo FAPESP: 08/50114-4 - Estudo do cupim Coptotermes gestroi: análise da base genética da diferenciação de castas, busca de genes de importância biotecnológica e estudo de celulases
Beneficiário:Fernando Ferreira Costa
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 10/11469-1 - Desenvolvimento de uma biblioteca de enzimas a partir de metagenoma de solo
Beneficiário:Thabata Maria Alvarez
Linha de fomento: Bolsas no Brasil - Doutorado Direto
Processo FAPESP: 08/58037-9 - Geração de biblioteca para conversão enzimática de biomassa a partir de metagenoma do solo
Beneficiário:Fábio Márcio Squina
Linha de fomento: Auxílio à Pesquisa - Programa BIOEN - Apoio a Jovens Pesquisadores