An alkaline active feruloyl-CoA synthetase from soil metagenome as a potential key enzyme for lignin valorization strategies

Author(s)

Sodre, Victoria ^{[1, 2]} ; Araujo, Juscemacia Nascimento ^[3] ; Goncalves, Thiago Augusto ^{[1, 4]} ; Vilela, Nathalia ^{[1, 4]} ; Kimus Braz, Antonio Sergio ^[3] ; Franco, Telma Teixeira ^[2] ; Neto, Mario de Oliveira ^[5] ; de Lima Damasio, Andre Ricardo ^[1] ; Garcia, Wanius ^[3] ; Squina, Fabio Marcio ^{[4, 2]}

Affiliation

^[1] Univ Campinas UNICAMP, Inst Biol, Dept Biochem & Tissue Biol, Campinas, SP - Brazil

^[2] Univ Campinas UNICAMP, Fac Chem Engn, Campinas, SP - Brazil

^[3] Univ Fed ABC UFABC, Ctr Ciencias Nat & Humanas, Santo Andre, SP - Brazil

^[4] Univ Sorocaba UNISO, Programa Proc Tecnol & Ambientais, Sorocaba, SP - Brazil

^[5] Univ Estadual Paulista UNESP, Inst Biociencias, Dept Fis & Biofis, Botucatu, SP - Brazil

• Document type: Journal article
• Source: PLoS One; v. 14, n. 2 FEB 25 2019.
• Web of Science Citations: 1

Abstract

Ferulic acid (FA), a low-molecular weight aromatic compound derived from lignin, represents a high-value molecule, used for applications in the cosmetic and pharmaceutical industries. FA can be further enzymatically converted in other commercially interesting molecules, such as vanillin and bioplastics. In several organisms, these transformations often start with a common step of FA activation via CoA-thioesterification, catalyzed by feruloyl-CoA synthetases (Fcs). In this context, these enzymes are of biotechnological interest for conversion of lignin-derived FA into high value chemicals. In this study, we describe the first structural characterization of a prokaryotic Fcs, named FCS1, isolated from a lignin-degrading microbial consortium. The FCS1 optimum pH and temperature were 9 and 37 degrees C, respectively, with Km of 0.12 mM and Vmax of 36.82 U/mg. The circular dichroism spectra indicated a notable secondary structure stability at alkaline pH values and high temperatures. This secondary structure stability corroborates the activity data, which remains high until pH 9. The Small Angle X-Ray Scattering analyses resulted on the tertiary/quaternary structure and the low-resolution envelope in solution of FCS1, which was modeled as a homodimer using the hyperthermophilic nucleoside diphosphate-forming acetyl-CoA synthetase from Candidatus Korachaeum cryptofilum. This study contributes to the field of research by establishing the first biophysical and structural characterization for Fcs, and our data may be used for comparison against novel enzymes of this class that to be studied in the future. (AU)

• FAPESP's process: 15/50612-8 - An integrated approach to explore a novel paradigm for biofuel production from lignocellulosic feedstocks
• Grantee: Telma Teixeira Franco
• Support type: Research Projects - 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 type: Scholarships in Brazil - Doctorate (Direct)
• 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 type: Scholarships in Brazil - Post-Doctorate
• FAPESP's process: 17/05901-7 - Development of E. coli-Rhodococcus CRISPR/Cas9 shuttle vector targeting ferulic acid accumulating Rhodococcus jostii RHA1 mutants
• Grantee: Victoria Ramos Sodré de Castro
• Support type: Scholarships in Brazil - Master
• FAPESP's process: 17/08166-6 - Genetic engineering of yeast for conversion lignocellulosic material to ferulic acid
• Grantee: Nathália Vilela
• Support type: Scholarships in Brazil - Doctorate
• 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 type: Regular Research Grants
• FAPESP's process: 17/22669-0 - N-glycosylation and enzymes secretion in filamentous fungi
• Grantee: André Ricardo de Lima Damasio
• Support type: Program for Research on Bioenergy (BIOEN) - Regular Program Grants
• FAPESP's process: 17/16976-8 - Biophysical and biochemical characterization of two bacterial enzymes Involved in the conversion of ferulic acid to vanillin
• Grantee: Juscemácia Nascimento Araujo
• Support type: Scholarships in Brazil - Doctorate
• 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 type: Program for Research on Bioenergy (BIOEN) - Thematic Grants