Applying biochemical and structural characterization of hydroxycinnamate catabolic enzymes from soil metagenome for lignin valorization strategies

Author(s):Show less -

Goncalves, Thiago Augusto ; Sodre, Victoria ; da Silva, Stephanie Nemesio ; Vilela, Nathalia ; Tomazetto, Geizecler ; Araujo, Juscemacia Nascimento ; Muniz, Joao Renato C. ; Fill, Taicia Pacheco ; Damasio, Andre ; Garcia, Wanius ; Squina, Fabio Marcio

• Document type: Journal article
• Source: Applied Microbiology and Biotechnology; v. 106, n. 7, p. 14-pg., 2022-03-30.

Abstract

The biocatalytic production of fuels and chemicals from plant biomass represents an attractive alternative to fossil fuel-based refineries. In this context, the mining and characterization of novel biocatalysts can promote disruptive innovation opportunities in the field of lignocellulose conversion and valorization. In the present work, we conducted the biochemical and structural characterization of two novel hydroxycinnamic acid catabolic enzymes, isolated from a lignin-degrading microbial consortium, a feruloyl-CoA synthetase, and a feruloyl-CoA hydratase-lyase, named LM-FCS2 and LM-FCHL2, respectively. Besides establishing the homology model structures for novel FCS and FCHL members with unique characteristics, the enzymes presented interesting biochemical features: LM-FCS2 showed stability in alkaline pHs and was able to convert a wide array of p-hydroxycinnamic acids to their respective CoA-thioesters, including sinapic acid; LM-FCHL2 efficiently converted feruloyl-CoA and p-coumaroyl-CoA into vanillin and 4-hydroxybenzaldehyde, respectively, and could produce vanillin directly from ferulic acid. The coupled reaction of LM-FCS2 and LM-FCHL2 produced vanillin, not only from commercial ferulic acid but also from a crude lignocellulosic hydrolysate. Collectively, this work illuminates the structure and function of two critical enzymes involved in converting ferulic acid into high-value molecules, thus providing valuable concepts applied to the development of plant biomass biorefineries. (AU)

• 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 Opportunities: Scholarships in Brazil - Doctorate
• 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: 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: 17/17275-3 - 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
• FAPESP's process: 17/22669-0 - N-glycosylation and enzymes secretion in filamentous fungi
• Grantee: André Ricardo de Lima Damasio
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Regular Program Grants
• 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 Opportunities: Research Projects - Thematic 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
• 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