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UNIVERSIDAD COMPLUTENSE DE MADRID FACULTAD DE CIENICAS QUÍMICAS Departamento De Bioquímica Y Biología Molecular UNIVERSIDAD COMPLUTENSE DE MADRID FACULTAD DE CIENICAS QUÍMICAS Departamento de Bioquímica y Biología Molecular TESIS DOCTORAL Estudio de xilanasas fúngicas para el aprovechamiento de la biomasa lignocelulósica Study of fungal xylanases for the exploitation of lignocellulosic biomass MEMORIA PARA OPTAR AL GRADO DE DOCTOR PRESENTADA POR Manuel José Nieto Domínguez Directoras María Jesús Martínez Hernández Laura Isabel de Eugenio Martínez Madrid, 2018 © Manuel José Nieto Domínguez, 2017 FACULTAD DE CIENCIAS QUÍMICAS DEPARTAMENTO DE BIOQUÍMICA Y BIOLOGÍA MOLECULAR I TESIS DOCTORAL ESTUDIO DE XILANASAS FÚNGICAS PARA EL APROVECHAMIENTO DE LA BIOMASA LIGNOCELULÓSICA STUDY OF FUNGAL XYLANASES FOR THE EXPLOITATION OF LIGNOCELLULOSIC BIOMASS MANUEL JOSÉ NIETO DOMÍNGUEZ DIRECTORES MARÍA JESÚS MARTÍNEZ HERNÁNDEZ LAURA ISABEL DE EUGENIO MARTÍNEZ MADRID, 2017 FACULTAD DE CIENCIAS QUÍMICAS DEPARTAMENTO DE BIOQUÍMICA Y BIOLOGÍA MOLECULAR I TESIS DOCTORAL ESTUDIO DE XILANASAS FÚNGICAS PARA EL APROVECHAMIENTO DE LA BIOMASA LIGNOCELULÓSICA STUDY OF FUNGAL XYLANASES FOR THE EXPLOITATION OF LIGNOCELLULOSIC BIOMASS MANUEL JOSÉ NIETO DOMÍNGUEZ DIRECTORES MARÍA JESÚS MARTÍNEZ HERNÁNDEZ LAURA ISABEL DE EUGENIO MARTÍNEZ MADRID, 2017 A mi abuelo Manolo A mis padres Tranquilo tío, somos científicos Bill Murray (como el Dr. Peter Venkman), Los Cazafantasmas ACKNOWLEDGMENTS This work has been carried out with funding from projects PRI-PIBAR-2011- 1402, RTC-2014-1777-3, BIO2015-68387-R from MINECO and RETO-PROSOT S2013/MAE-2972 from Comunidad de Madrid. Manuel Nieto thanks the MINECO for his FPU fellowship and the Department of Biochemistry and Molecular Biology I (Faculty of Chemistry, Universidad Complutense de Madrid) for the teaching training. El que aquestas palabras suscribe es tristemente consciente de que, en el espacio que se le ha concedido, difícilmente podrá dar cumplida cuenta de todos aquellos que se han hecho merecedores de su gratitud durante estos venturosos cuatro años. Mas su honra empeña en que intentará dar a cada uno lo que en justicia le correspondiere y si alguno se perdiere en el tintero de esta torpe pluma, sepa que aunque la razón haya fallado en el corazón permanece. En primer lugar no puedo sino dar las gracias a la directora de esta tesis, la Dra. María Jesús Martínez, gran científica y mejor persona. Te doy las gracias por confiar en mí todos y cada uno de los días que he pasado en tu laboratorio, por reírte con mis exposiciones (sin despedirme luego), por tu optimismo a prueba de ataques nucleares, por tu guía y ánimos, por enseñarme hacer las cosas bien y por tu mágica habilidad para solucionar el problema más complicado en el último minuto. En co-primer lugar gracias a la Dra. Laura de Eugenio, con toda probabilidad la mejor codirectora que haya existido jamás, considerando el planeta Tierra y varios universos paralelos. Dueña de mi destino, en la salud y en la enfermedad, hasta que la tesis nos separe, pero amiga para siempre. Gracias por tu humor negro y tu cariño incondicional pobremente disimulado. Gracias por enseñarme que aunque las cosas no hayan salido del todo bien, siempre hay alguien que te compra la burra. Gracias por la tarta Guinness y hasta por el heavy metal (no, eso es broma). En general, gracias por todo Lidem. Gracias a todos mis compañeros de Biotecnología para la Biomasa Lignocelulósica, ¿quién iba a pensar que detrás de un nombre tan aburrido hubiera tan buena gente? Gracias a Jezú, embajador de Barbate en el mundo y mi cicerone en el laboratorio. Gracias por tu buen humor, porque contigo aprendí que aunque los experimentos no salgan, el Madrid o Nadal acaban por ganar algo y por supuesto gracias por incumplir siempre tu amenaza de pegarme. Gracias pisha. Gracias a Mariu, fuente y destino de toda información. Mi vecina de poyata, compañera de tantas penas y alguna alegría. Gracias por tu paciencia, por enseñarme lo que es un salseo en condiciones y ante todo por estar siempre ahí, dispuesta a ayudarme cada vez que lo necesitaba. Gracias Davichúa Salvachúa por tu alegría contagiosa y tu sabiduría enciclopédica. Aprendí de ti desde el día que llegué hasta el que te fuiste a vivir el sueño americano. Gracias a Peces, Rosa Peces, insultantemente joven, pero fichaje estrella del laboratorio. Gracias por ser tan buena gente, por poner orden en el caos cuando nos quedamos sin Mariu (sólo han pasado cuatro líneas y ya te echo de menos otra vez Mariu), por tus ganas de reír y en definitiva por ser un sol. Gracias a Juanillo (A.), hermano de hongo. Gracias por esos cafés de avellana y ciencia, por tu coche siempre listo para una misión de los Manolos, por ser un Caballero de la Lamprea e inasequible al desaliento. Que sepas que si no te para tu tobillo no te parará nadie. ¿Y qué decir de Fran? Pues que llegó a mi vida como una ola de fuerza desmedida y cargada de medios, tampones y placas siempre a punto. El cielo es más azul desde que tú estás en el laboratorio y aguanta despejado hasta las cinco de la tarde. No te vayas nunca. Gracias a Úrsula, inventora de la capoeira. Si el 249 fuese una serie tú serías mi personaje recurrente preferido. Gracias a Norge, el Dr. Quorum Sensing. Gracias por recorrer los duros caminos de la ciencia y haber vuelto entero (más o menos) para contárnoslos y gracias por esas reuniones en el Instituto Ondiñas de Investigación de firmes comienzos y borrosos finales. Gracias a María, de personalidad magnética y repostera inigualable (tú también eres inigualable Lidem). Gracias por ser la voz de la razón en el loco país de Nunca Jamás. Gracias a Neumara, perfecto equilibrio entre samba brasileña e ingeniería química. Gracias porque metodología de respuesta en superficie suena (un poco) más ameno con acento tropical. Gracias a la Dra. Ali, co-co-directora en la sombra e ilustre políglota. Gracias por tus ganas de pasarlo bien haciendo ciencia de primera, porque tu buen humor es como la contaminación en Madrid, no hay forma de perderlo y lo más importante, por tener siempre cinco minutos para sentarte conmigo aunque haya una multitud con antorchas en tu puerta. Gracias a Marijose, jefa absoluta del laboratorio, que me enseñó todo lo que un hombre necesita saber (y lo que no necesita también) sobre ligar en el bingo y besos sorpresa. Cambio de laboratorio y también de párrafo que empezaba a ser demasiado largo. Gracias a Lola Linde López (L3), capaz de iluminar una habitación con su sonrisa mientras nos intoxica con tolueno. Gracias a Remedios Pacheco, más conocida como Piña, gracias por tu risa perenne y por enseñarme que el arte moderno no es (tan) malo. Gracias a Vero, un ángel con pipetas, en Suecia no saben la suerte que tienen. Gracias a Juanillo (R.), compañero gallego en el exilio, tu cara de sueño y resignación por las mañanas me hacía sentir menos sólo en la vida. Gracias a Ana, exportadora del mañacao, y posiblemente la postdoc más joven de espíritu (lo que no es espíritu quedará en el misterio) que haya conocido. Gracias a Iván, el latin lover de Ávila, gracias por todos esos buenos consejos que nunca seguí sin que dejaras de ser mi amigo. Gracias por las apuestas absurdas de incierto resultado y sobre todo gracias por convencerme para que, de vez en cuando, disfrutara de la luz solar. Gracias a David, Amanolito para los amigos y ¡Almendral! para los más íntimos. Gracias por ser genio y figure y por liarla lo justo para desviar de nosotros la atención de los jefes. Como sé que leerás esta frase contando las palabras que te he dedicado continuaré alargando innecesariamente el texto, por lo menos una línea más, para asegurarme de que quedas total y absolutamente satisfecho. Gracias Supernieto. Gracias también a Ángel Martínez, sapientísimo doctor y al locuaz Dr. Javi Ruíz Dueñas por mantener los euros de Europa fluyendo, la impresora en marcha y acertar siempre con los licores de sobremesa. Gracias a Falip, Pilaf al revés, alquimista de la anilina y el sumial. Gracias por ser un romántico incomprendido y buen compañero donde los haya. Gracias a Isa (P), encantadora y temible a partes iguales, pero brillante como nadie. Gracias por acoger pacientemente el rol en el salón de tu casa. Gracias a Isa (V), mi amor platónico (platónico de momento) y la persona más luminosa del CIB en particular y del Consejo Superior de Investigaciones Científicas en general. Gracias porque allá donde estás más claros los fluorescentes brillan y los solventes orgánicos se respiran mejor. Gracias a la Dra. Susana Camarero, la ambición rubia y la única persona capaz de hacer divertido hasta un congreso de lignina. Gracias a todos aquellos que en algún momento han pasado por el laboratorio aportándome siempre algo positivo. Gracias a Cris Coscolín, por ser locamente genial (algunos dirían genialmente loca), a Lorea que me descubrió que hay comidas veganas que saben hasta bien, a Erik, el del gorro perenne, a Bea Balcells, Elena, Aitor, Jose, Lucía, los Felipes Pavel, Jacobo, David y Mario (tú también eres un Felipe, Mario y lo sabes) y a alguno más que probablemente me esté olvidando, pero que así podrá echármelo en cara en el futuro. Sin salir del CIB pero cambiando de planta tengo que dar las gracias a mis compañeros de RMN, los Dres. Javier Cañada y Ángeles Canales y muy especialmente a Bea capaz de resolver un xilósido en menos de 24 horas y de desaparearme los espines con una sola mirada encantadora. Gracias también a los Dres. Antonio Romero y Elena Santillana por ayudarme a asomar la mirada al arduo mundo de la cristalografía de rayos X. Tampoco me puedo olvidar de Blas Blázquez (me podría poner un warning), maestro de las magics, ni de Juan Nogals, el Señor Oscuro que el día menos pensado te puede dar trabajo.
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