Recombinant expression, purification and characterization of an active bacterial feruloyl-CoA synthase with potential for application in vanillin production

Author(s)

dos Santos, Otavio Augusto Leitao ; Goncalves, Thiago Augusto ; Sodre, Victoria ; Vilela, Nathalia ; Tomazetto, Geizecler ; Squina, Fabio M. ; Garcia, Wanius

• Document type: Journal article
• Source: Protein Expression and Purification; v. 197, p. 7-pg., 2022-05-11.

Abstract

The ferulic acid (FA) represents a high-value molecule with applications in the cosmetic and pharmaceutical industries. This aromatic molecule is derived from lignin and can be enzymatically converted in other commercially interesting molecules, such as vanillin and bioplastics. This process starts with a common step of FA activation via CoA-thioesterification, catalyzed by feruloyl-CoA synthetases. Therefore, here, we report the successfully expression, purification as well as the initial structural and biochemical characterization of a stable, correctly folded, and catalytically active bacterial feruloyl-CoA synthase (here named FCS3) isolated from a lignin-degrading microbial consortium. The purification of recombinant FCS3 to near homogeneity was achieved using affinity chromatography. The FCS3 structure is composed of a mixture of alpha and beta secondary structures and most likely forms stable homodimers in solution. The FCS3 presented a notable structural stability at alkaline pH values and it was able to convert FA and coenzyme A (CoA) into feruloyl-CoA complex at room temperature. This study should provide a useful basis for future biotechnological applications of FCS3, especially in the field of conversion of lignin-derived FA into high value compounds. (AU)

• FAPESP's process: 17/16089-1 - Production of lignin chains, metagenomic screening and biocatalytic conversion of lignin to ferulic acid
• Grantee: Thiago Augusto Gonçalves
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• 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
• FAPESP's process: 18/18101-1 - Development of the CRISPR/Cas9 system in Rhodococcus jostii RHA1 for production of high-value chemicals from lignin streams
• Grantee: Victoria Ramos Sodré de Castro
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 17/08166-6 - Genetic engineering of yeast for conversion lignocellulosic material to ferulic acid
• Grantee: Nathália Vilela
• Support Opportunities: Scholarships in Brazil - Doctorate
• 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: 17/17275-3 - Studies of the mode of action of two lytic polysaccharide monooxygenases from insect (order: isoptera): molecular structure, bioinorganic chemistry and biotechnological applications
• Grantee: Wanius José Garcia da Silva
• Support Opportunities: Regular Research Grants