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

Author(s)

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

• Document type: Journal article
• Source: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS; v. 1865, n. 4, p. 395-403, APR 2017.
• Web of Science Citations: 12

Abstract

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)

• FAPESP's process: 09/52840-7 - Center of Biological and Industrial Processes for Biofuels - CeProBIO
• Grantee: Igor Polikarpov
• Support type: Program for Research on Bioenergy (BIOEN) - Thematic Grants
• FAPESP's process: 13/21545-5 - Identification and molecular studies of enzymes from thermophilic fungi Aspergillus niveus and Myceliophtora thermophila
• Grantee: Fernando Segato
• Support type: Scholarships in Brazil - Post-Doctorate
• FAPESP's process: 14/18714-2 - Enzymatic oxidation of sugarcane bagasse: discovery, characterization and new application of oxidative enzymes active in carbohydrates, applied to the enhancement of a fungal cell factory
• Grantee: Fernando Segato
• Support type: Program for Research on Bioenergy (BIOEN) - Young Investigators Grants