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

• Published in: Protein expression and purification Vol. 197; p. 106109
• Main Authors: dos Santos, Otávio Augusto Leitão, Gonçalves, Thiago Augusto, Sodré, Victoria, Vilela, Nathalia, Tomazetto, Geizecler, Squina, Fabio M., Garcia, Wanius
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.09.2022
• Subjects: Coenzyme A; Ferulic acid; Feruloyl-CoA synthase; Vanillin; Coenzyme A; Vanillin; Feruloyl-CoA synthase; Ferulic acid
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2022.106109

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 α and β 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. •The ferulic acid (FA) can be enzymatically converted in vanillin.•Feruloyl-CoA synthase with potential for application in vanillin production.•Biochemical and structural characterization of a bacterial feruloyl-CoA synthase.•Feruloyl-CoA synthase was able to convert FA and CoA into feruloyl-CoA complex.