The scientific objectives of the group are related to the use of microorganisms (mainly filamentous fungi) and their enzymes in industrial processes to obtain fuels, materials and chemicals (White Biotechnology) from renewable plant resources. The final aim is to contribute to the sustainable development of our society and reduce the biosphere warming by a reduced consumption of fossil resources.
Hydrolytic enzymes for industrial and environmental applications
The research line led by Mª Jesús Martinez aims to understand key aspects of the catalytic action of fungal hydrolases involved in the biodegradation or biotransformation of lignocelluloses and lipids, emphasizing on versatile lipases, cellulases and hemicellulases, and includes the laboratories of Alicia Prieto, who has high expertise in carbohydrate chemistry, and Jorge Barriuso working on quorum sensing mechanisms and related topics. These basic aspects entail in-depth biochemical, physic-chemical and structural characterization of the enzymes, structure-function studies, and use of computational and bioinformatics tools.
The outcomes from these experimental approaches are essential to explain and improve the catalytic and/or operational properties of the proteins when subjected to industrial and environmental applications based on the exploitation of residual plant biomass and other wastes as renewable feedstock from agriculture, industry and other human activities. These tasks are currently gathered in two projects, HIDROFUN and RETO-PROSOST.
The most relevant biotechnological applications of our research are related to the enzymatic production of second generation bioethanol and biodiesel and added-value products, as prebiotic oligosaccharides, lipid or sugar derivatives with enhanced solubility and biological activity, flavors and fragrances. In all of them, biocatalysis replaces chemical reagents, sometimes releases products that cannot be synthesized chemically, and contributes to the development of green and sustainable processes.
Engineering of oxidative enzymes
The research line led by Angel T. Martínez is related to protein (redox enzymes) design and their utilization in oxidative chemical reactions of industrial interest, as those explored in the INDOX (www.indoxproject.eu), EnzOx2 (http://enzox2.eu), WoodZymes (https://woodzymes.eu/) and SusBind (https://susbind.eu/) EU projects, and includes the laboratories of Susana Camarero, working on enzyme directed evolution, and Francisco Javier Ruiz-Dueñas, with high expertise in fungal multiomics. Our work in these projects focuses on proteins of the (i) heme-peroxidase, (ii) multicopper oxidases and (iii) flavo-oxidase types, as Green Chemistry catalysts for different oxidation and oxyfunctionalization reactions of industrial interest.
The enzyme engineering work takes advantage from: i) Computational simulations of protein-ligand interactions (computational biophysics) and quantum mechanics/molecular mechanics (QM/MM) to predict the electron transfer (computational biochemistry), in a well-established collaboration with the Barcelona Supercomputing Center (www.bsc.es); ii) Analytical chemistry at CIB (MS, NMR, etc) and in collaboration with IRNAS in Seville (www.irnas.csic.es); and iii) Genomic information on fungi responsible for "enzymatic combustion" of lignin (the second more abundant macromolecule on earth) including projects with the Joint Genome Institute (http://jgi.doe.gov) like CSP-2015-1609, coordinated from CIB, for analyzing 32 fungal genomes.
|Ángel Tomás Martínez Ferrer|
|María Jesús Martínez Hernández|
|Alicia María Prieto Orzanco|
|Francisco Javier Ruiz Dueñas|
|Susana Camarero Fernández|
|Juan Rogelio Carro Aramburu|
|Marta Pérez Boada|
|María Dolores Línde López|
|Ivan Ayuso Fernandez|
|Ana Serrano Esteban|
|Jorge Barriuso Maicas|
|Laura Isabel de Eugenio Martínez|
|Mihai Lucian Lomanar|
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Viñambres M, Espada M, Martínez AT, Serrano A . Screening and Evaluation of New Hydroxymethylfurfural Oxidases for Furandicarboxylic Acid Production. Appl. Environ. Microbiol. 86:e00842-20
Babot ED, Aranda C, del Río JC, Ullrich R, Kiebist J, Scheibner K, Hofrichter M, Martínez AT, Gutiérrez A . Selective Oxygenation of Ionones and Damascones by Fungal Peroxygenases. J. Agric. Food Chem., DOI: 10.1021/acs.jafc.0c01019
Linde D, Olmedo A, González-Benjumea A, Estévez M, Renau-Mínguez C, Carro J, Fernández-Fueyo E, Gutiérrez A, Martínez AT . Two New Unspecific Peroxygenases from Heterologous Expression of Fungal Genes in Escherichia coli. Appl. Environ. Microbiol. 86:e02899-19
Schneider WDH, Fontana RC, Baudel HM, Gonçalves de Siqueira F, Rencoret J, Gutiérrez A, de Eugenio LI, Prieto A, Martínez MJ, Martínez AT, Pinheiro Dillona AJ, Camassolaa M . Lignin degradation and detoxification of eucalyptus wastes by on-site manufacturing fungal enzymes to enhance second-generation ethanol yield. Applied Energy 262:114493
González-Benjumea A, Carro J, Renau-Mínguez C, Linde D, Fernández-Fueyo E, Gutiérrez A, Martínez AT . Fatty acid epoxidation by Collariella virescens peroxygenase and heme-channel variants. Catal. Sci. Technol. 10: 717–725
de Salas F, Aza P, Gilabert JF, Santiago G, Kilic S, Sener ME, Vind J, Guallar V, Martínez AT and Camarero S . Engineering of a fungal laccase to develop a robust, versatile and highly-expressed biocatalyst for sustainable chemistry. Green Chemistry doi: 10.1039/c9gc02475a
de Salas F, Cañadas R, Santiago G, Virseda-Jerez A, Vind J, Gentili P, Martínez AT, Guallar V, Muñoz IG, Camarero S . Structural and biochemical insights into an engineered high-redox potential laccase overproduced in Aspergillus. Int. J. Biol. Macromol. doi: 10.1016/j.ijbiomac.2019.09.052
Serrano A, Calviño E, Carro J, Sánchez-Ruiz MI, Cañada FJ, Martínez AT . Complete oxidation of hydroxymethylfurfural to furandicarboxylic acid by aryl-alcohol oxidase. Biotechnol. Biofuels, 12: 127
Ayuso-Fernández I, Rencoret J, Gutiérrez A, Ruiz-Dueñas FJ, Martínez AT . Peroxidase evolution in white-rot fungi follows wood lignin evolution in plants. Proc. Natl. Acad. Sci. USA, doi: 10.1073/pnas.1905040116
Romero JO, Fernandez-Fueyo E, Avila-Salas F, Recabarren R, Alzate-Morales J, Martínez AT . Binding and Catalytic Mechanisms of Veratryl Alcohol Oxidation by Lignin Peroxidase: A Theoretical and Experimental Study. Comput. Struct. Biotechnol. J ., 17: 1066-1074
Schneider WDH, Bolaño Losada C, Moldes D, Fontana RC, Gonçalves de Siqueira F, Prieto A, Martínez MJ, Martínez AT, Pinheiro Dillon AJ, Camassola M . A Sustainable Approach of Enzymatic Grafting on Eucalyptus globulus Wood by Laccase from the Newly Isolated White-Rot Basidiomycete Marasmiellus palmivorus VE111. ACS Sustainable Chem. Eng. 7:13418−13424
Carro J, González-Benjumea A, Fernández-Fueyo E, Aranda C, Guallar V, Gutiérrez A, Martínez AT . Modulating Fatty Acid Epoxidation vs Hydroxylation in a Fungal Peroxygenase. ACS Catal. 9: 6234-6242
Méndez-Líter JA, Tundidor I, Nieto-Domínguez M, Fernández de Toro B, González Santana A, de Eugenio LI, Prieto A, Asensio JL, Cañada FJ, Sánchez C and Martínez MJ . Transglycosylation products generated by Talaromyces amestolkiae GH3 β-glucosidases: effect of hydroxytyrosol, vanillin and its glucosides on breast cancer cells. Microb. Cell Fact. 18:97
Payá-Tormo L, Rodríguez-Salarichs J, Prieto A, Martínez MJ, Barriuso J . Improvement of the Activity of a Fungal Versatile-Lipase Toward Triglycerides: An in silico Mechanistic Description. Front. Bioeng. Biotechnol. 7:71
The public (including EU, MINECO-Spain and CAM-Madrid) and private (contracts with companies) financial sources are included in the list provided below, which includes: i) Project or Contract title, ii) Responsible (or coordinator); iii) Funding; iv) Funding entity and reference; and v) Duration.
A. The projects currently in course are listed below:
1) "Glycosyl hydrolases and other auxiliary enzymes for the production of added value compounds" (GLYSUS) PI: MJ Martinez; co-PI: A Prieto, Ministerio Ciencia e Innovación (RTI2018-093683-B-I00, 2019-21); Funding CIB: 162.000 € (2019-2021)
2) "Sustainable production and industrial symbiosis in the Community of Madrid" (RETO-PROSOST-2-CM) PIs: A Prieto/MJ Martínez, Comunidad de Madrid (P2018/EMT4459, 2019-2022)
3) “Extremozymes for wood based building blocks: From pulp mill to board and insulation products” (WoodZymes) EC H2020, Coordinator S. Camarero, Ref H2020-BBI-JTI-2017-792070, Total funding 3253873 €; CIB 738340 € (2018-21), www.woodzymes.eu
4) “Development and pilot production of SUStainable bio BINDer systems for wood based panels” (SusBind), EC H2020, PI-CIB: A.T. Martínez, Ref H2020-BBI-JTI-2017-792063; Total funding 4414418 €; CIB 756170 € (2018-22), www.susbind.eu
5) "Basidiomycete genomes for lignocellulose biorefineries" (GENOBIOREF), PIs F.J. Ruiz-Dueñas/S. Camarero, MINECO/FEDER (BIO2017-86559-R) Funding 302500 € (2018-22)
6) “Biotechnology for lignocellulose: towards the integral use of plant biomass as renewable resource”. Acciones de dinamización “Redes de Excelencia”. MINECO. Ref. BIO2017-90757-REDT, Funding: 15000 €; 2018 -2020. PI: S. Camarero. www.redlignocel.es
7) “Industrial Biotechnology Innovation and Synthetic Biology Accelerator” (IBISBA 1.0) EC H2020; PI-CIB: M.J. Martínez; Ref. H2020-INFRAIA-730976 Funding CIB: 275.000 € (2017-21) www.ibisba.eu
8) "Fungal hydrolases: biochemical and molecular studies and applications for lignocellulosic waste revalorization" (HIDROFUN), PIs M.J. Martínez/A. Prieto, MINECO/FEDER (BIO2015-68387-R), Funding 199650 € (2016-18)
9) “New enzymatic oxidation/oxyfunctionalization technologies for added value bio-based products” (EnzOx2), EC H2020, Coordinator A.T. Martínez, PI-CIB F.J. Ruiz-Dueñas, Ref H2020-BBI-PPP-2015-2-720297, Total funding 3000000 €; CIB 606000 € (2016-19), www.enzox2.eu
B. Other relevant (EU or large National) previous projects on related topics:
"Sustainable production and industrial symbiosis in the Community of Madrid" (RETO-PROSOST) PI: MJ Martinez, Comunidad de Madrid (S2013/MAE-2907), Total funding 694950 €, CIB 179860 € (2014-2018)
"New oxidative enzymes for sustainable industries" (NOESIS), PIs: S. Camarero/F.J. Ruiz-Dueñas, MINECO (BIO2014-56388-R). Funding: 302500 € (2015-18)
"Enzymatic, chemical and engineering challenges for the use of non-food agroforestal resources (lignocellulose) in a more sustainable and environmentally-friendly bioeconomy" (network LIGNOCEL), Coordinator S. Camarero, Ref AC2015-00008-00-00, funding 6000 € (2016), www.redlignocel.es
"Optimized oxidoreductases for medium and large scale industrial biotransformations" (INDOX), EC FP7, Coordinator A.T. Martínez, PI-CIB F.J. Ruiz-Dueñas, Ref KBBE-2013-7-613549, total funding 7825824 €: CIB 928654 € (2013-16), www.indoxproject.eu
"Combined approaches based on metagenomics for biofuels synthesis" (CAMBIOS), PI M.J. Martínez, MINECO (RTC2014-1777-3). Funding: 95742 € (2014-17)
"Novel and more robust fungal peroxidases as industrial biocatalysts" (PEROXICATS), FP7, Coordinator A.T. Martínez, PI-CIB F.J. Ruiz-Dueñas, Ref KBBE-2010-4-265397, total funding 2 999 350 euros: CIB 732 114 euros, 2011-13, www.peroxicats.org
“Optimised pre-treatment of fast growing woody and nonwoody Brazilian crops by detailed characterisation of chemical changes produced in the lignin-carbohydrate matrix” (LIGNODECO), FP7, PI-CSIC A.T. Martínez, Ref KBBE-2009-3-244362, total funding 2 988 220 euros; CIB 451 250 euros, 2010-2012
"High-performance biocatalysts for second generation bioethanol" (BIOCAT2ndOL), one of the six sub-projects of the project on "Production and development of non-food energetic cultures" asigned to CSIC by the MICINN (as a part of the Spanish PlanE) with a total budeget of 8 512 507 euros; PI-CIB M.J. Martínez; 2009-2011
"Ethanol for automotion" (I+DEA); Coordinator: P. Gutiérrez (Abengoa Bioenergy); CIB-PI: M.J. Martínez; total budget (including contributions from 25 companies) 28 million euros; CIB funding 646 000 euros; CDTI, CENIT 2007-1031; 2007-2011
“White biotechnology for added value products from renewable plant polymers: Design of tailor-made biocatalysts and new industrial bioprocesses” (BIORENEW); A.T. Martínez (coordinator); total budget: 14 683 137 euros: total EU contribution: 9 500 000 euros; EU contribution to CIB: 1 012 388 euros; Priority-3 (NMP) of EU FP6, NMP2-CT-2006-026456; 2006-2010, www.biorenew.org