Enzymatic and biophysical characterization of a novel modular cellulosomal GH5 endoglucanase multifunctional from the anaerobic gut fungus Piromyces finnis

Author(s)

Andrade, Viviane Brito ; Tomazetto, Geizecler ; Almeida, Dnane Vieira ; Tramontina, Robson ; Squina, Fabio Marcio ; Garcia, Wanius

• Document type: Journal article
• Source: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS; v. 1872, n. 1, p. 11-pg., 2024-01-01.

Abstract

Cellulases from anaerobic fungi are enzymes less-studied biochemically and structurally than cellulases from bacteria and aerobic fungi. Currently, only thirteen GH5 cellulases from anaerobic fungi were biochemically characterized and two crystal structures were reported. In this context, here, we report the functional and biophysical characterization of a novel multi-modular cellulosomal GH5 endoglucanase from the anaerobic gut fungus Piromyces finnis (named here PfGH5). Multiple sequences alignments indicate that PfGH5 is composed of a GH5 catalytic domain and a CBM1 carbohydrate-binding module connected through a CBM10 dockerin module. Our results showed that PfGH5 is an endoglucanase from anaerobic fungus with a large spectrum of activity. PfGH5 exhibited preference for hydrolysis of oat beta-glucan, followed by galactomannan, carboxymethyl cellulose, mannan, lichenan and barley beta-glucan, therefore displaying multi-functionality. For oat beta-glucan, PfGH5 reaches its optimum enzymatic activity at 40 degrees C and pH 5.5, with Km of 7.1 mu M. Ion exchange chromatography analyzes revealed the production of oligosaccharides with a wide degree of polymerization indicated that PfGH5 has endoglucanase activity. The ability to bind and cleave different types of carbohydrates evidence the potential of PfGH5 for use in biotechnology and provide a useful basis for future investigation and application of new anaerobic fungi enzymes. (AU)

• FAPESP's process: 20/15595-3 - Functional and structural study of a cellulosomal cellulase from anaerobic fungus and its dockerin-scaffoldin interaction
• Grantee: Wanius José Garcia da Silva
• Support Opportunities: Regular Research Grants
• FAPESP's process: 20/05784-3 - EMU approved in grant 15 / 50590-4: chromatographic system and detectors for analysis of sugars and lignocellulosic monolignols
• Grantee: Fábio Márcio Squina
• Support Opportunities: Multi-user Equipment Program
• FAPESP's process: 21/04254-3 - Development of Biorefinery concepts via coumaric acid conversion into high-value compounds
• Grantee: Robson Tramontina
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 15/23279-6 - Metagenomic and metasecretomic analyses of an enrichment microbial consortium of anaerobic fungi to the degradation of sugarcane bagasse: prospection of cellulosomas and lignocellolytic enzymes
• Grantee: Geizecler Tomazetto
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 22/08958-8 - Development of microorganisms and thermophilic enzymes involved in the degradation and upcycling of fossil plastics
• Grantee: Fábio Márcio Squina
• Support Opportunities: Regular Research Grants
• FAPESP's process: 15/50590-4 - Lignin valorization in cellulosic ethanol plants: biocatalytic conversion via ferulic acid to high value chemicals
• Grantee: Fábio Márcio Squina
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Thematic Grants