Cloning, heterologous expression and biochemical characterization of a non-specific endoglucanase family 12 from Aspergillus terreus NIH2624

Autor(es)

Segato, Fernando ; Dias, Bruno ; Berto, Gabriela L. ; de Oliveira, Dyoni M. ; De Souza, Flavio H. M. ; Citadini, Ana Paula ; Murakami, Mario T. ; Damasio, Andre R. L. ; Squina, Fabio Marcio ; Polikarpov, Igor

• Tipo de documento: Artigo Científico
• Fonte: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS; v. 1865, n. 4, p. 395-403, APR 2017.
• Citações Web of Science: 12

Resumo

The cellulases from Glycoside Hydrolyses family 12 (GH12) play an important role in cellulose degradation and plant cell wall deconstruction being widely used in a number of bioindustrial processes. Aiming to contribute toward better comprehension of these class of the enzymes, here we describe a high-yield secretion of a endoglucanase GH12 from Aspegillus terreus (AtGH12), which was cloned and expressed in Aspergillus nidulans strain A773.The purified protein was used for complete biochemical and functional characterization. The optimal temperature and pH of the enzyme were 55 degrees C and 5.0 respectively, which has high activity against beta-glucan and xyloglucan and also is active toward glucomannan and CMC. The enzyme retained activity up to 60 degrees C. AtGH12 is strongly inhibited by Cu2+, Fe2+, Cd2+, Mn2+, Ca2+, Zn2+ and EDTA, whereas K+, Tween, Cs+, DMSO, Triton X100 and Mg2+ enhanced the enzyme activity. Furthermore, SAXS data reveal that the enzyme has a globular shape and CD analysis demonstrated a prevalence of a beta-strand structure corroborating with typical beta-sheets fold commonly found for other endoglucanases from GH12 family. (C) 2017 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 13/21545-5 - Identificação e estudos moleculares de enzimas produzidas pelos fungos termofílicos Aspergillus niveus e Myceliophtora termophila
• Beneficiário: Fernando Segato
• Linha de fomento: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/18714-2 - Oxidação enzimática do bagaço de cana-de-açúcar: descoberta, caracterização e aplicação de novas enzimas oxidativas de carboidratos, voltadas para o desenvolvimento de um cell factory mais eficiente
• Beneficiário: Fernando Segato
• Linha de fomento: Auxílio à Pesquisa - Programa BIOEN - Apoio a Jovens Pesquisadores
• Processo FAPESP: 09/52840-7 - Centro de Processos Biológicos e Industriais para Biocombustíveis - CeProBIO
• Beneficiário: Igor Polikarpov
• Linha de fomento: Auxílio à Pesquisa - Programa BIOEN - Temático