Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture

Author(s):Show less -

Pimentel, Agnes C. ; Ematsu, Gabriela C. G. ; Liberato, Marcelo V. ; Paixao, Douglas A. A. ; Franco Cairo, Joao Paulo L. ; Mandelli, Fernanda ; Tramontina, Robson ; Gandin, Cesar A. ; de Oliveira Neto, Mario ; Squina, Fabio M. ; Alvarez, Thabata M.

• Document type: Journal article
• Source: International Journal of Biological Macromolecules; v. 99, p. 384-393, JUN 2017.
• Web of Science Citations: 5

Abstract

Endoglucanases are key enzymes in the degradation of cellulose, the most abundant polymer on Earth. The aim of this work was to perform the biochemical and biophysical characterization of CelE2, a soil metagenome derived endoglucanase. CelE2 harbors a conserved domain from glycoside hydrolase family 5 (GH5) and a C-terminal domain with identity to Calx-beta domains. The recombinant CelE2 displayed preference for hydrolysis of oat beta-glucan, followed by lichenan and carboxymethyl cellulose. Optimum values of enzymatic activity were observed at 45 degrees C and pH 5.3, and CelE2 exhibited considerable thermal stability at 40 degrees C for up to 360 min. Regarding the cleavage pattern on polysaccharides, the release of oligosaccharides with a wide degree of polymerization indicated a characteristic of endoglucanase activity. Furthermore, the analysis of products generated from the cleavage of cellooligosaccharides suggested that CelE2 exhibited transglycosylation activity. Interestingly, the presence of CaCl2 positively affect CelE2, including in the presence of surfactants. SAXS experiments provided key information on the effect of CaCl2 on the stability of CelE2 and dummy atom and rigid-body models were generated. To the best of our knowledge this is the first biochemical and biophysical characterization of an endoglucanase from family GH5 displaying this unconventional modular organization. (C) 2017 Elsevier B.V. All rights reserved. (AU)

• FAPESP's process: 14/06923-6 - Sugar cane biomass recalcitrance: basic knowledge related to the cell wall construction, pretreatment and enzymatic digestion, applied for the development of innovative biorefinery models
• Grantee: Andre Luis Ferraz
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Thematic Grants
• FAPESP's process: 14/04105-4 - Structural and functional characterization of new cellulases, focusing in the relation between catalytic domains and CBMs
• Grantee: Marcelo Vizoná Liberato
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 16/01926-2 - Influence of an accessory domain in biochemical and structural characteristics of the cellulase CelE2
• Grantee: Agnes Cristina Pimentel
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 10/11469-1 - Library generation for biomass-conversion enzymes from soil metagenome
• Grantee: Thabata Maria Alvarez
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 14/12861-3 - Biochemical characterization of a metagenomic cellulase from family 5 and determination of biotechnological application
• Grantee: Agnes Cristina Pimentel
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 08/58037-9 - Library generation for biomass-conversion enzymes from soil metagenome
• Grantee: Fábio Márcio Squina
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Young Investigators Grants