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Lignin valorization in cellulosic ethanol plants: biocatalytic conversion via ferulic acid to high value chemicals

Grant number: 15/50590-4
Support Opportunities:Program for Research on Bioenergy (BIOEN) - Thematic Grants
Duration: June 01, 2017 - January 31, 2023
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Convênio/Acordo: BBSRC, UKRI
Principal Investigator:Fábio Márcio Squina
Grantee:Fábio Márcio Squina
Principal researcher abroad: Tim David Howard Bugg
Institution abroad: University of Warwick, England
Host Institution: Pró-Reitoria de Pós-Graduação, Pesquisa, Extensão e Inovação. Universidade de Sorocaba (UNISO). Sorocaba , SP, Brazil
Associated grant(s):20/05784-3 - EMU approved in grant 15 / 50590-4: chromatographic system and detectors for analysis of sugars and lignocellulosic monolignols, AP.EMU
Associated scholarship(s):22/09248-4 - Conversion of lignin to ferulic acid (WP2), BP.TT
22/05117-2 - WP1 - Integration between lignin valorization and cellulosic ethanol production, BP.TT
21/14783-3 - Conversion of lignin to ferulic acid (WP2), BP.TT
+ associated scholarships 21/04254-3 - Development of Biorefinery concepts via coumaric acid conversion into high-value compounds, BP.PD
21/13886-3 - Bioprocesses intensification, expansion of chemical products production and lignocellulose processing, BP.TT
21/08564-7 - Conversion of lignin to ferulic acid (WP2), BP.TT
20/15483-0 - Biocatalytic production of high-chemical from lignin streams, BP.TT
20/14543-0 - WP3 - Biocatalytic formation of fragrance chemicals from ferulic acid, BP.TT
20/11019-8 - WP1: integration between lignin valorization and cellulosic ethanol production, BP.TT
20/03474-7 - WP3: biocatalytic formation of fragrance chemicals from ferulic acid, BP.TT
20/03051-9 - Bioprocesses intensification, expansion of chemical products production and lignocellulose processing, BP.TT
19/26321-4 - Conversion of lignin to ferulic acid (WP2), BP.TT
18/18101-1 - Development of the CRISPR/Cas9 system in Rhodococcus jostii RHA1 for production of high-value chemicals from lignin streams, BP.DD
17/08166-6 - Genetic engineering of yeast for conversion lignocellulosic material to ferulic acid, BP.DR
17/16089-1 - Production of lignin chains, metagenomic screening and biocatalytic conversion of lignin to ferulic acid, BP.PD
17/05901-7 - Development of E. coli-Rhodococcus CRISPR/Cas9 shuttle vector targeting ferulic acid accumulating Rhodococcus jostii RHA1 mutants, BP.MS
16/07926-4 - Novel integrative strategies through the aldoketo reductase from the termite Coptotermes gestroi for detoxification, saccharification and fermentation of lignocellulose aiming bioethanol production, BP.DR
16/09950-0 - Functional, structural characterizations and biotechnological application of lytic polysaccharide monooxygenases from the lower termite Coptotermes gestroi, BP.PD - associated scholarships

Abstract

Lignin can be obtained as a by-product of cellulosic ethanol production, and is a potential source of renewable chemicals, if methods for lignin valorization can be developed. Efficient valorisation of lignin is a major unsolved problem in the development of sustainable biorefineries. The proposal builds upon an existing BBSRC partnership award, and brings together expertise in biocatalyst discovery and lignocelullose ethanol production (CTBE) with expertise in biocatalytic lignin valorization (Warwick) and biocatalysis for high value chemicals production (Manchester, UCL) The overall aim is to generate new methods for lignin valorization via intermediate ferulic acid, which has been generated from lignin in previous Pl's works. The project will involve the following work packages: 1) optimization of lignin generation from cellulosic bioethanol; 2) conversion of lignin to ferulic acid from lignin using synthetic biology; 3) enzymatic conversion of ferulic acid into pharmaceutical L-Dopa; 4) biocatalytic generation of high value fragrance chemicals (coniferyl acetate, isoeugenol) from ferulic acid; 5) bioprocess and scale-up of chemicals production from renewable feedstocks; 6) technical and sustainability impact assessment. Technology developed in the project could be applied to major plant feedstocks used in Brazil (sugar cane) or the UK (wheat). (AU)

Articles published in Pesquisa para Inovação FAPESP about research grant:
Novel biotechnological route developed to obtain fine chemicals from agricultural waste 
Articles published in Agência FAPESP Newsletter about the research grant:
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Scientific publications (32)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DE FIGUEIREDO, FERNANDA LOPES; DE OLIVEIRA, ANA CAROLINA PIVA; TERRASAN, CESAR RAFAEL FANCHINI; GONCALVES, THIAGO AUGUSTO; GERHARDT, JAQUELINE ALINE; TOMAZETTO, GEIZECLER; PERSINOTI, GABRIELA FELIX; RUBIO, MARCELO VENTURA; PENA, JENNIFER ANDREA TAMAYO; ARAUJO, MICHELLE FERNANDES; et al. Multi-omics analysis provides insights into lignocellulosic biomass degradation by Laetiporus sulphureus ATCC 52600. BIOTECHNOLOGY FOR BIOFUELS, v. 14, n. 1, . (19/21609-0, 15/50612-8, 17/22669-0, 20/05784-3, 19/12860-0, 15/50590-4)
SODRE, VICTORIA; VILELA, NATHALIA; TRAMONTINA, ROBSON; SQUINA, FABIO MARCIO. icroorganisms as bioabatement agents in biomass to bioproducts application. BIOMASS & BIOENERGY, v. 151, . (16/07926-4, 18/18101-1, 15/50590-4, 17/08166-6)
DA SILVA, VIVIAM M.; CABRAL, ALINE D.; SPERANCA, MARCIA A.; SQUINA, FABIO M.; MUNIZ, JOAO RENATO C.; MARTIN, LYDIE; NICOLET, YVAIN; GARCIA, WANIUS. High-resolution structure of a modular hyperthermostable endo-beta-1,4-mannanase from Thermotoga petrophila: The ancillary immunoglobulin-like module is a thermostabilizing domain. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1868, n. 8, . (13/26096-4, 16/15836-5, 17/17275-3, 17/16291-5, 14/02065-5, 16/14514-4, 14/50897-0, 15/50590-4)
BRENELLI, LIVIA B.; PERSINOTI, GABRIELA F.; FRANCO CAIRO, JOAO PAULO L.; LIBERATO, MARCELO V.; GONCALVES, THIAGO AUGUSTO; OTERO, IGOR V. R.; MAINARDI, PEDRO H.; FELBY, CLAUS; SETTE, LARA D.; SQUINA, FABIO M.. Novel redox-active enzymes for ligninolytic applications revealed from multiomics analyses of Peniophora sp. CBMAI 1063, a laccase hyper-producer strain. SCIENTIFIC REPORTS, v. 9, . (13/03061-0, 13/19486-0, 15/07008-2, 14/04105-4, 16/09950-0, 15/06971-3, 15/50590-4, 17/16089-1)
FRANCO CAIRO, JOAO PAULO L.; CANNELLA, DAVID; OLIVEIRA, LEANDRO C.; GONCALVES, THIAGO A.; RUBIO, MARCELO V.; TERRASAN, CESAR R. F.; TRAMONTINA, ROBSON; MOFATTO, LUCIANA S.; CARAZZOLLE, MARCELO F.; GARCIA, WANIUS; et al. On the roles of AA15 lytic polysaccharide monooxygenases derived from the termite Coptotermes gestroi. Journal of Inorganic Biochemistry, v. 216, . (17/17275-3, 16/09950-0, 15/50590-4, 17/11952-3, 15/50612-8, 17/22669-0, 17/16089-1)
CORREA, THAMY L. R.; FRANCO CAIRO, JOAO PAULO L.; COTA, JUNIO; DAMASIO, ANDRE; OLIVEIRA, LEANDRO C.; SQUINA, FABIO M.. A novel mechanism of beta-glucosidase stimulation through a monosaccharide binding-induced conformational change. International Journal of Biological Macromolecules, v. 166, p. 1188-1196, . (15/50590-4, 16/09950-0, 12/20549-4, 10/18198-3)
TRAMONTINA, ROBSON; BRENELLI, LIVIA BEATRIZ; SOUSA, AMANDA; ALVES, RAFAEL; ZETTY ARENAS, ANA MARIA; NASCIMENTO, VIVIANE MARCOS; RABELO, SARITA CANDIDA; FREITAS, SINDELIA; RULLER, ROBERTO; SQUINA, FABIO MARCIO. Designing a cocktail containing redox enzymes to improve hemicellulosic hydrolysate fermentability by microorganisms. Enzyme and Microbial Technology, v. 135, . (15/50590-4, 13/03061-0, 17/15477-8, 15/26982-0, 16/07926-4)
BRENELLI, LIVIA BEATRIZ; MARIUTTI, LILIAN REGINA BARROS; PORTUGAL, RODRIGO VILLARES; DE FARIAS, MARCELO ALEXANDRE; BRAGAGNOLO, NEURA; MERCADANTE, ADRIANA ZERLOTTI; FRANCO, TELMA TEIXEIRA; RABELO, SARITA CANDIDA; SQUINA, FABIO MARCIO. odified lignin from sugarcane bagasse as an emulsifier in oil-in-water nanoemulsion. INDUSTRIAL CROPS AND PRODUCTS, v. 167, . (13/03061-0, 20/05784-3, 15/50612-8, 15/50590-4)
ALVAREDO PAIXAO, DOUGLAS ANTONIO; TOMAZETTO, GEIZECLER; SODRE, VICTORIA RAMOS; GONCALVES, THIAGO A.; UCHIMA, CRISTIANE AKEMI; BUCHLI, FERNANDA; ALVAREZ, THABATA MARIA; PERSINOTI, GABRIELA FELIX; DA SILVA, MARCIO JOSE; BRAGATTO, JULIANO; et al. Microbial enrichment and meta-omics analysis identify CAZymes from mangrove sediments with unique properties. Enzyme and Microbial Technology, v. 148, . (12/19040-0, 15/50590-4, 14/04105-4, 16/09950-0, 15/23279-6, 18/18101-1, 17/16089-1)
SPENCE, EDWARD M.; SCOTT, HEATHER T.; DUMOND, LOUISON; CALVO-BADO, LEONIDES; DI MONACO, SABRINA; WILLIAMSON, JAMES J.; PERSINOTI, GABRIELA F.; SQUINA, FABIO M.; BUGG, TIMOTHY D. H.. The Hydroxyquinol Degradation Pathway in Rhodococcus jostii RHA1 and Agrobacterium Species Is an Alternative Pathway for Degradation of Protocatechuic Acid and Lignin Fragments. Applied and Environmental Microbiology, v. 86, n. 19, . (15/50590-4)
DOS SANTOS, OTAVIO AUGUSTO LEITAO; GONCALVES, THIAGO AUGUSTO; SODRE, VICTORIA; VILELA, NATHALIA; TOMAZETTO, GEIZECLER; SQUINA, FABIO M.; GARCIA, WANIUS. Recombinant expression, purification and characterization of an active bacterial feruloyl-CoA synthase with potential for application in vanillin production. Protein Expression and Purification, v. 197, p. 7-pg., . (17/16089-1, 15/50590-4, 18/18101-1, 17/08166-6, 20/15595-3, 20/05784-3, 17/17275-3)
MARTINS, MANOELA; TRAMONTINA, ROBSON; SQUINA, FABIO MARCIO; DINAMARCO, TAISA MAGNANI; GOLDBECK, ROSANA. Synergism for xylo-oligosaccharides, p-coumaric and ferulic acid production, and thermostability modulation of GH 62 alpha-L-arabinofuranosidase. BIOCATALYSIS AND AGRICULTURAL BIOTECHNOLOGY, v. 44, p. 11-pg., . (20/02871-2, 15/50590-4, 19/08542-3)
PIMENTEL, AGNES C.; LIBERATO, MARCELO V.; CAIRO, JOAO PAULO L. FRANCO; TOMAZETTO, GEIZECLER; GANDIN, CESAR A.; NETO, MARIO DE OLIVEIRA; ALVAREZ, THABATA M.; SQUINA, FABIO M.. Structural and functional insights of the catalytic GH5 and Calx-beta domains from the metagenome-derived endoglucanase CelE2. Enzyme and Microbial Technology, v. 165, p. 10-pg., . (16/01926-2, 15/50590-4, 20/05784-3, 14/04105-4, 10/11469-1, 16/09950-0, 15/23279-6)
TRAMONTINA, ROBSON; CIANCAGLINI, IARA; ROMAN, ELLEN K. B.; CHACON, MICAELA G.; CORREA, THAMY L. R.; DIXON, NEIL; BUGG, TIMOTHY D. H.; SQUINA, FABIO MARCIO. Sustainable biosynthetic pathways to value-added bioproducts from hydroxycinnamic acids. Applied Microbiology and Biotechnology, v. 107, n. 13, p. 21-pg., . (15/50590-4, 21/04254-3)
ANDRADE, VIVIANE BRITO; TOMAZETTO, GEIZECLER; ALMEIDA, DNANE VIEIRA; TRAMONTINA, ROBSON; SQUINA, FABIO MARCIO; GARCIA, WANIUS. Enzymatic and biophysical characterization of a novel modular cellulosomal GH5 endoglucanase multifunctional from the anaerobic gut fungus Piromyces finnis. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1872, n. 1, p. 11-pg., . (20/15595-3, 20/05784-3, 21/04254-3, 15/23279-6, 22/08958-8, 15/50590-4)
PERSINOTI, GABRIELA F.; PAIXAO, DOUGLAS A. A.; BUGG, TIMOTHY D. H.; SQUINA, FABIO M.. Genome Sequence of Lysinibacillus sphaericus, a Lignin-Degrading Bacterium Isolated from Municipal Solid Waste Soil. GENOME ANNOUNCEMENTS, v. 6, n. 18, p. 2-pg., . (15/50590-4)
GRANJA-TRAVEZ, ROMMEL SANTIAGO; WILKINSON, RACHAEL C.; PERSINOTI, GABRIELA FELIX; SQUINA, FABIO M.; FULOP, VILMOS; BUGG, TIMOTHY D. H.. Structural and functional characterisation of multi-copper oxidase CueO from lignin-degrading bacterium Ochrobactrum sp reveal its activity towards lignin model compounds and lignosulfonate. FEBS Journal, v. 285, n. 9, p. 1684-1700, . (15/50590-4)
MORAES, EDUARDO C.; ALVAREZ, THABATA M.; PERSINOTI, GABRIELA F.; TOMAZETTO, GEIZECLER; BRENELLI, LIVIA B.; PAIXAO, DOUGLAS A. A.; EMATSU, GABRIELA C.; ARICETTI, JULIANA A.; CALDANA, CAMILA; DIXON, NEIL; et al. Lignolytic-consortium omics analyses reveal novel genomes and pathways involved in lignin modification and valorization. BIOTECHNOLOGY FOR BIOFUELS, v. 11, . (13/03061-0, 14/26152-4, 15/50590-4, 15/23279-6)
LIMA, CLEILTON S.; RABELO, SARITA C.; CIESIELSKI, PETER N.; ROBERTO, INES C.; ROCHA, GEORGE J. M.; DRIEMEIER, CARLOS. Multiscale Alterations in Sugar Cane Bagasse and Straw Submitted to Alkaline Deacetylation. ACS SUSTAINABLE CHEMISTRY & ENGINEERING, v. 6, n. 3, p. 3796-3804, . (15/50590-4)
PERSINOTI, GABRIELA F.; PAIXAO, DOUGLAS A. A.; BUGG, TIMOTHY D. H.; SQUINA, FABIO M.. Genome Sequence of Lysinibacillus sphaericus, a Lignin-Degrading Bacterium Isolated from Municipal Solid Waste Soil. MICROBIOLOGY RESOURCE ANNOUNCEMENTS, v. 6, n. 18, . (15/50590-4)
TRAMONTINA, ROBSON; GALMAN, JAMES L.; PARMEGGIANI, FABIO; DERRINGTON, SASHA R.; BUGG, TIMOTHY D. H.; TURNER, NICHOLAS J.; SQUINA, FABIO M.; DIXON, NEIL. Consolidated production of coniferol and other high-value aromatic alcohols directly from lignocellulosic biomass. GREEN CHEMISTRY, v. 22, n. 1, p. 144-152, . (15/50590-4, 17/23754-1)
TRAMONTINA, ROBSON; BRENELLI, LIVIA BEATRIZ; SODRE, VICTORIA; FRANCO CAIRO, JOAO PAULO; TRAVALIA, BEATRIZ MEDEIROS; EGAWA, VIVIANE YOSHIMI; GOLDBECK, ROSANA; SQUINA, FABIO MARCIO. Enzymatic removal of inhibitory compounds from lignocellulosic hydrolysates for biomass to bioproducts applications. WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY, v. 36, n. 11, . (20/03051-9, 15/50590-4, 16/07926-4, 17/15477-8, 18/18101-1)
MOREIRA, EDIMAR A.; PERSINOTI, GABRIELA F.; MENEZES, LETICIA R.; PAIXAO, DOUGLAS A. A.; ALVAREZ, THABATA M.; CAIRO, JOAO P. L. FRANCO; SQUINA, FABIO M.; COSTA-LEONARDO, ANA MARIA; RODRIGUES, ANDRE; SILLAM-DUSSES, DAVID; et al. Complementary Contribution of Fungi and Bacteria to Lignocellulose Digestion in the Food Stored by a Neotropical Higher Termite. FRONTIERS IN ECOLOGY AND EVOLUTION, v. 09, . (15/21497-6, 16/09950-0, 18/22839-6, 15/50590-4)
GRANATO, DANIELA C.; NEVES, LEANDRO X.; TRINO, LUCIANA D.; CARNIELLI, CAROLINA M.; LOPES, ARIANE F. B.; YOKOO, SAMI; PAULETTI, BIANCA A.; DOMINGUES, ROMENIA R.; SA, JAMILE O.; PERSINOTI, GABRIELLA; et al. Meta-omics analysis indicates the saliva microbiome and its proteins associated with the prognosis of oral cancer patients. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1869, n. 8, . (18/18496-6, 18/02180-0, 16/07846-0, 19/18751-9, 19/21815-9, 09/54067-3, 18/11958-4, 15/50590-4, 18/12194-8, 18/15728-3, 14/06485-9, 18/15535-0, 10/19278-0)
PUHL, ANA C.; PRATES, ERICA T.; ROSSETO, FLAVIO R.; MANZINE, LIVIA R.; STANKOVIC, IVANA; DE ARAUJO, SIMARA S.; ALVAREZ, THABATA M.; SQUINA, FABIO M.; SKAF, MUNIR S.; POLIKARPOV, IGOR. Crystallographic structure and molecular dynamics simulations of the major endoglucanase from Xanthomonas campestris pv. campestris shed light on its oligosaccharide products release pattern. International Journal of Biological Macromolecules, v. 136, p. 493-502, . (11/20505-4, 15/13684-0, 09/52840-7, 10/52362-5, 11/21608-1, 15/50590-4)
TOMAZETTO, GEIZECLER; PIMENTEL, AGNES C.; WIBBERG, DANIEL; DIXON, NEIL; SQUINA, FABIO M.. Multi-omic Directed Discovery of Cellulosomes, Polysaccharide Utilization Loci, and Lignocellulases from an Enriched Rumen Anaerobic Consortium. Applied and Environmental Microbiology, v. 86, n. 18, . (16/01926-2, 18/23826-5, 15/50590-4, 15/23279-6)
LIBERATO, MARCELO VIZONA; CAMPOS, BRUNA MEDEIA; TOMAZETTO, GEIZECLER; CROUCH, LUCY ISOBEL; GARCIA, WANIUS; DE MATTOS ZERI, ANA CAROLINA; BOLAM, DAVID NICHOL; SQUINA, FABIO MARCIO.

Unique properties of a Dictyostelium discoideum carbohydrate-binding module expand our understanding of CBM-ligand interactions

. Journal of Biological Chemistry, v. 298, n. 5, p. 10-pg., . (15/50590-4, 13/06336-0, 15/23279-6, 20/15595-3, 14/04105-4, 20/05784-3, 15/14009-5, 17/17275-3)
VELASCO, JOSMAN; OLIVA, BIANCA; GONCALVES, ALINE LARISSA; LIMA, AWANA SILVA; FERREIRA, GISLENE; FRANCA, BRUNO ALVES; MULINARI, EVANDRO JOSE; GONCALVES, THIAGO AUGUSTO; SQUINA, FABIO MARCIO; SEIKI KADOWAKI, MARCO ANTONIO; et al. Functional characterization of a novel thermophilic exo-arabinanase fromThermothielavioides terrestris. Applied Microbiology and Biotechnology, v. 104, n. 19, . (14/18714-2, 17/16089-1, 14/06923-6, 19/01165-0, 15/50590-4, 19/06663-8)
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; et al. Applying biochemical and structural characterization of hydroxycinnamate catabolic enzymes from soil metagenome for lignin valorization strategies. Applied Microbiology and Biotechnology, v. 106, n. 7, p. 14-pg., . (17/16976-8, 20/05784-3, 18/18101-1, 17/17275-3, 17/22669-0, 15/50612-8, 15/50590-4, 15/23279-6)
FANCHINI TERRASAN, CESAR RAFAEL; RUBIO, MARCELO VENTURA; GERHARDT, JAQUELINE ALINE; FRANCO CAIRO, JOAO PAULO; CONTESINI, FABIANO JARES; ZUBIETA, MARIANE PALUDETTI; DE FIGUEIREDO, FERNANDA LOPES; VALADARES, FERNANDA LIMA; RIBEIRO CORREA, THAMY LIVIA; MURAKAMI, MARIO TYAGO; et al. Deletion of AA9 Lytic Polysaccharide Monooxygenases Impacts A. nidulans Secretome and Growth on Lignocellulose. MICROBIOLOGY SPECTRUM, v. N/A, p. 16-pg., . (15/50612-8, 15/50590-4, 17/22669-0, 16/16306-0)
SODRE, VICTORIA; ARAUJO, JUSCEMACIA NASCIMENTO; GONCALVES, THIAGO AUGUSTO; VILELA, NATHALIA; KIMUS BRAZ, ANTONIO SERGIO; FRANCO, TELMA TEIXEIRA; NETO, MARIO DE OLIVEIRA; DE LIMA DAMASIO, ANDRE RICARDO; GARCIA, WANIUS; SQUINA, FABIO MARCIO. An alkaline active feruloyl-CoA synthetase from soil metagenome as a potential key enzyme for lignin valorization strategies. PLoS One, v. 14, n. 2, . (17/05901-7, 17/22669-0, 17/16976-8, 15/50612-8, 17/16089-1, 15/50590-4, 17/08166-6, 18/18101-1, 17/17275-3)
VALADARES, FERNANDA; GONCALVES, THIAGO A.; DAMASIO, ANDRE; MILAGRES, ADRIANE M. F.; SQUINA, FABIO M.; SEGATO, FERNANDO; FERRAZ, ANDRE. The secretome of two representative lignocellulose-decay basidiomycetes growing on sugarcane bagasse solid-state cultures. Enzyme and Microbial Technology, v. 130, . (17/16089-1, 15/50590-4, 14/06923-6)

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