The scientific objectives of our group include:
1) Providing knowledge on the biological decay of plant biomass with special emphasis on the transformation and degradation of lignin by fungi, a key process for C recycling in nature and the complete exploitation of plant biomass as raw material in the Lignocellulose Biorefineries, as an alternative to the use of fossil resources.
2) Promoting the use of these organisms and their enzymes (improved in the lab by molecular biology, protein engineering, computational design and synthetic biology techniques) for the industrial conversion of renewable plant biomass into bio-based chemicals and/or materials, including the synthesis of bioplastics.
3) Contributing in this way to the concept of Circular (Bio)economy through more sustainable, efficient and clean industrial processes (and with lower carbon footprint) in which organic waste (agricultural, forestry or industrial) is not generated or is used, as well as to the valorization and recycling of oil-derived plastic polymers.
To achieve the above objectives, we develop basic and applied research lines. The former focus on unveiling the composition and understanding the behaviour of the enzymatic machinery responsible for the transformation of lignocellulose by fungi specialized in its degradation (projects CSP15-1609 from DOE-JGI and GenoBioref from BIO2017). To do this, we carry out multi-omic studies, including secretomic, transcriptomic and comparative genomic analyses of a wide variety of saprophytic fungal species, complemented with chemical analyses of degraded substrates. These studies are key for the discovery of new enzymes. We are particularly interested in oxidoreductases involved in the biodegradation of the lignin polymer and other aromatic compounds of great biotechnological interest. Of special interest are also the peroxygenases secreted by saprophytic fungi, homologous to (intracellular) P450s but phylogenetically unrelated. Both share the ability to selectively catalyze oxygenation reactions that are difficult to achieve by chemical means, but while P450s require auxiliary proteins and a source of reducing power, the fungal peroxygenases are self-sufficient and require only peroxide to be activated. This fact, together with their greater robustness as secreted enzymes, provides them a much higher biotechnological potential as selective biocatalysts in the chemical sector. Finally, the phylogenetic and evolutionary studies of all these enzymes, and the computational reconstruction and subsequent resurrection and characterization of ancestral enzymes, provide valuable information to understand the changes responsible for the properties of current enzymes and for the design of new tailor-made enzymes of industrial interest.
The applied research lines start from the heterologous expression of the most interesting extant or resurrected oxidoreductases from a biotechnological point of view. These studies focus on improving the enzyme properties or acquiring the new functionalities required (for each specific application) by means of directed evolution and/or rational design, as well as their use as biocatalysts in oxidative synthesis or degradation reactions of industrial interest, such as those investigated in the WoodZymes (https://woodzymes.eu), SusBind (https://susbind.eu), and FurEnPol (PLEC2021-007690) projects, led by our group. Our work in these projects focuses on enzymes of the (i) multicopper oxidase, (ii) heme-peroxidase, (iii) heme-peroxygenase, and (iv) flavo-oxidase types, and their use as biocatalysts in the conversion of plant materials in value-added compounds or in components of bio-based products, developing oxidation and oxyfunctionalization reactions of interest in Green Chemistry. The group also participates in the CSIC SusPlast Interdisciplinary Thematic Platform on "Sustainable Plastics for a Circular Economy", with the aim of contributing with biotechnological solutions to the production of bio-based plastics from renewable sources and the transformation and recycling of petroleum-derived polymers.
The activities carried out by our group are aligned with several of the global change challenges proposed in the CSIC 2025 Strategy. In this sense, we actively contribute to finding answers to scientific challenges with a high social impact, such as those included in the WoodZymes, SusBind and FurEnPol mentioned above. In the same way, our research work is reflected in patents and first-level high-impact international publications, as a result of interdisciplinary collaborations, which have allowed us to consolidate collaborations with national, European and American universities, research centers and companies.
|Susana Camarero Fernández|
|Ángel Tomás Martínez Ferrer|
|Francisco Javier Ruiz Dueñas|
|Marta Pérez Boada|
|María Dolores Línde López|
|Juan Rogelio Carro Aramburu|
|Pablo Aza Toca|
|Gonzalo Molpeceres García|
|María Isabel Sánchez Ruíz|
|David Rodriguez Escribano|
|Juan Carral Saez-Royuela|
|Alba Saez Matia|
|Jose Manuel Gonzalez Fornell|
|Isabel Ocampos Crespo|
|Rodrigo Rincon Sanz|
|Manuel Garcia-Cervigon Plaza|
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Linde D, González-Benjumea A, Aranda C, Carro J, Gutiérrez A and Martínez AT . Engineering Collariella virescens Peroxygenase for Epoxides Production from Vegetable Oil. Antioxidants 11:915
Linde L, Santillana E, Fernández-Fueyo E, González-Benjumea A, Carro J, Gutiérrez A, Martínez AT and Romero A . Structural Characterization of Two Short Unspecific Peroxygenases: Two Different Dimeric Arrangements. Antioxidants 11:891
Olmedo A, Ullrich R, Hofrichter M, del Río JC, Martínez AT and Gutiérrez A . Novel Fatty Acid Chain-Shortening by Fungal Peroxygenases Yielding 2C-Shorter Dicarboxylic Acids. Antioxidants 11:744
Babot E, Aranda C, Kiebist J, Scheibner K, Ullrich R, Hofrichter M, Martinez AT and Gutierrez A . Enzymatic Epoxidation of Long-Chain Terminal Alkenes by Fungal Peroxygenases. Antioxidants 11:522
González-Benjumea A, Linde D, Carro J, Ullrich R, Hofrichter M, Martínez AT and Gutiérrez A . Regioselective and Stereoselective Epoxidation of n-3 and n-6 Fatty Acids by Fungal Peroxygenases. Antioxidants 10:1888
Rencoret J, Gutiérrez A, Marques G, del Río JC, Tobimatsu Y, Lam PY, Pérez-Boada M, Ruiz-Dueñas FJ, Barrasa JM and Martínez AT . New Insights on Structures Forming the Lignin-Like Fractions of Ancestral Plants. Front. Plant Sci. doi: 10.3389/fpls.2021.740923
Sánchez-Ruiz MI, Ayuso-Fernández I, Rencoret J, González-Ramírez AM, Linde D, Davó-Siguero I, Romero A, Gutiérrez A, Martínez AT and Ruiz-Dueñas FJ . Agaricales Mushroom Lignin Peroxidase: From Structure–Function to Degradative Capabilities. Antioxidants 10:1446
Sánchez-Ruiz MI, Martínez AT and Serrano A . Optimizing operational parameters for the enzymatic production of furandicarboxylic acid building block. Microb. Cell Fact. 20:180
de Salas F, Camarero S . Fungal Laccases as Biocatalysts for Wide Range Applications. In: Zaragoza, O. (ed) Encyclopedia of Mycology. vol. 2, pp. 233–246. Oxford: Elsevier
Peña A, Babiker R, Chaduli D, Lipzen A, Wang M, Chovatia M, Rencoret J, Marques G, Sánchez-Ruiz MI, Kijpornyongpan T, Salvachúa D, Camarero S, Ng V, Gutiérrez A, Grigoriev IV, Rosso M-N, Martínez AT and Ruiz-Dueñas FJ . A Multiomic Approach to Understand How Pleurotus eryngii Transforms Non-Woody Lignocellulosic Material. J. Fungi 7:426
Aza P, Molpeceres G, Ruiz-Dueñas FJ and Camarero S . Heterologous Expression, Engineering and Characterization of a Novel Laccase of Agrocybe pediades with Promising Properties as Biocatalyst. J. Fungi 7:359
Aza P, Molpeceres G, de Salas F and Camarero S . Design of an improved universal signal peptide based on the α-factor mating secretion signal for enzyme production in yeast. Cell Mol. Life Sci. 78:3691–3707
Linde D, Ayuso-Fernández I, Laloux M, Aguiar-Cervera JE, de Lacey AL, Ruiz-Dueñas FJ and Martínez AT . Comparing Ligninolytic Capabilities of Bacterial and Fungal Dye-Decolorizing Peroxidases and Class-II Peroxidase-Catalases. Int. J. Mol. Sci. 22:2629
Aza P, de Salas F, Molpeceres G, Rodríguez-Escribano D, de la Fuente I, Camarero S . Protein Engineering Approaches to Enhance Fungal Laccase Production in S. cerevisiae. Int. J. Mol. Sci. 22:1157
The public EU and national financial sources are included in the list provided below, which includes: i) Project title, ii) Responsible (or coordinator); iii) Funding; iv) Funding entity and reference; and v) Duration.
A. The funded projects currently in course are listed below:
1) “Enzymatic Synthesis and Recycling of Biobased Furanic Polymers” (FURENPOL), PIs: AT Martínez/FJ Ruiz-Dueñas, R&D&I Projects on Strategic Lines – 2021, Ref. PLEC2021-007690/AEI/10.13039/501100011033, Total funding: 1.038.039 €; CIB Funding: 310.406 € (2021-2024)
2) “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
3) “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
4) "Basidiomycete genomes for lignocellulose biorefineries" (GENOBIOREF), PIs F.J. Ruiz-Dueñas/S. Camarero, MINECO/FEDER (BIO2017-86559-R) Funding 302500 € (2018-22)
5) “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
B. Other relevant EU or National previous projects on related topics:
“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
"New oxidative enzymes for sustainable industries" (NOESIS), PIs: S. Camarero/F.J. Ruiz-Dueñas, MINECO (BIO2014-56388-R). Funding: 302500 € (2015-18)
"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
"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, https://cordis.europa.eu/project/id/244362/es
“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, https://cordis.europa.eu/project/id/26456/es