Structure and Function of a Novel Cellulase 5 from Sugarcane Soil Metagenome

Autor(es):Mostrar menos -

Alvarez, Thabata M. ^[1] ; Paiva, Joice H. ^[2] ; Ruiz, Diego M. ^[2] ; Cairo, Joao Paulo L. F. ^[1] ; Pereira, Isabela O. ^[1] ; Paixao, Douglas A. A. ^[1] ; de Almeida, Rodrigo F. ^[1] ; Tonoli, Celisa C. C. ^[2] ; Ruller, Roberto ^[1] ; Santos, Camila R. ^[2] ; Squina, Fabio M. ^[1] ; Murakami, Mario T. ^[2]

Afiliação do(s) autor(es)

^[1] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, Campinas, SP - Brazil

^[2] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Biociencias LNBio, Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PLoS One; v. 8, n. 12 DEC 17 2013.
• Citações Web of Science: 21

Resumo

Cellulases play a key role in enzymatic routes for degradation of plant cell-wall polysaccharides into simple and economically-relevant sugars. However, their low performance on complex substrates and reduced stability under industrial conditions remain the main obstacle for the large-scale production of cellulose-derived products and biofuels. Thus, in this study a novel cellulase with unusual catalytic properties from sugarcane soil metagenome (CelE1) was isolated and characterized. The polypeptide deduced from the celE1 gene encodes a unique glycoside hydrolase domain belonging to GH5 family. The recombinant enzyme was active on both carboxymethyl cellulose and beta-glucan with an endo-acting mode according to capillary electrophoretic analysis of cleavage products. CelE1 showed optimum hydrolytic activity at pH 7.0 and 50 degrees C with remarkable activity at alkaline conditions that is attractive for industrial applications in which conventional acidic cellulases are not suitable. Moreover, its three-dimensional structure was determined at 1.8 angstrom resolution that allowed the identification of an insertion of eight residues in the beta 8-alpha 8 loop of the catalytic domain of CelE1, which is not conserved in its psychrophilic orthologs. This 8-residue-long segment is a prominent and distinguishing feature of thermotolerant cellulases 5 suggesting that it might be involved with thermal stability. Based on its unconventional characteristics, CelE1 could be potentially employed in biotechnological processes that require thermotolerant and alkaline cellulases. (AU)

• Processo FAPESP: 11/20977-3 - Investigação de vias de degradação de biomassa lignocelulósica no cupim Coptotermes gestroi complementares a hidrolases glicolíticas visando aplicação na produção de bioprodutos derivados de biomassa lignocelulósica
• Beneficiário: João Paulo Lourenço Franco Cairo
• Linha de fomento: Bolsas no Brasil - Doutorado
• Processo FAPESP: 13/13309-0 - Estudos do comportamento funcional e estrutural de enzimas evolutivamente especializadas na degradação de biomassa vegetal com potenciais aplicações biotecnológicas
• Beneficiário: Mário Tyago Murakami
• 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: 09/08312-6 - Investigações funcionais e estruturais dos mecanismos moleculares envolvidos na sinalização, regulação e ativação das miosinas Classe V
• Beneficiário: Mário Tyago Murakami
• Linha de fomento: Auxílio à Pesquisa - Regular
• 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