{"id":24507,"date":"2003-09-07T00:00:00","date_gmt":"2003-09-07T00:00:00","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/expresion-heterologa-y-estudios-estructura-funcion-de-una-nueva-peroxidasa-versatil-de-pleurotus-eryngii\/"},"modified":"2003-09-07T00:00:00","modified_gmt":"2003-09-07T00:00:00","slug":"expresion-heterologa-y-estudios-estructura-funcion-de-una-nueva-peroxidasa-versatil-de-pleurotus-eryngii","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/expresion-heterologa-y-estudios-estructura-funcion-de-una-nueva-peroxidasa-versatil-de-pleurotus-eryngii\/","title":{"rendered":"Expresi\u00f3n heter\u00f3loga y estudios estructura-funci\u00f3n de una nueva peroxidasa vers\u00e1til de pleurotus eryngii"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Marta P\u00e9rez Boada <\/strong><\/h2>\n<p>En esta tesis se han abordado estudios estructura-funci\u00f3n de un nuevo tipo de peroxidasa producida por el hongo ligninol\u00edtico pleurotus eryngii, denominada peroxidasa vers\u00e1til (vp). Esta hemoprote\u00edna se caracteriza por su amplia especificidad de sustrato, que incluye desde el mn2+ hasta compuestos arom\u00e1ticos fen\u00f3licos y no fen\u00f3licos o colorantes de tipo azo. Se ha desarrollado un sistema de expresi\u00f3n heter\u00f3loga de la vp en escherichia coli, situando el cdna correspondiente a la prote\u00edna madura en el vector de expresi\u00f3n pflag1. La prote\u00edna expresada se acumul\u00f3 en forma de cuerpos de inclusi\u00f3n, que se aislaron y solubilizaron en presencia de urea, para realizar posteriormente el plegamiento in vitro de la peroxidasa recombinante.  se investig\u00f3 la influencia de diferentes par\u00e1metros en la reacci\u00f3n de plegamiento, como el ph, las condiciones redox y la concentraci\u00f3n de prote\u00edna, urea e iones calcio.  el protocolo optimizado permite recuperar hasta un 10% de enzima en forma activa, un rendimiento tres veces superior al obtenido para otras peroxidasas estructuralmente similares. Asimismo se han realizado estudios mediante la t\u00e9cnica de stopped-flow para la caracterizaci\u00f3n espectrosc\u00f3pica del ciclo catal\u00edtico y el an\u00e1lisis de la cin\u00e9tica del estado transitorio de la reacci\u00f3n de la vp con el h2o2 y el alcohol veratr\u00edlico (va), que es uno de sus sustratos reductores.  los resultados obtenidos ponen de manifiesto unas propiedades de los intermediarios de oxidaci\u00f3n (compuestos i y ii) muy similares a las de otras peroxidasas estudiadas. Se ha calculado la constante de formaci\u00f3n del compuesto i a distintos valores de ph, as\u00ed como las constantes de formaci\u00f3n y reducci\u00f3n del compuesto ii por el va.  los estudios de nmr de la prote\u00edna en disoluci\u00f3n y difracci\u00f3n de rayos x tras obtenci\u00f3n de cristales proporcionaron informaci\u00f3n estructural precisa de la vp. Los espectros de nmr de la vp mostraron que el entorno d<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Expresi\u00f3n heter\u00f3loga y estudios estructura-funci\u00f3n de una nueva peroxidasa vers\u00e1til de pleurotus eryngii<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Expresi\u00f3n heter\u00f3loga y estudios estructura-funci\u00f3n de una nueva peroxidasa vers\u00e1til de pleurotus eryngii <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Marta P\u00e9rez Boada <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Complutense de Madrid<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 09\/07\/2003<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>Direcci\u00f3n y tribunal<\/h3>\n<ul>\n<li><strong>Director de la tesis<\/strong>\n<ul>\n<li>Angel Tomas Martinez Ferrer<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: julian Perera gonzalez <\/li>\n<li>guillermo Gimenez gallego (vocal)<\/li>\n<li>Manuel Acosta echevarr\u00eda (vocal)<\/li>\n<li> Pe\u00f1alva soto Miguel angel (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Marta P\u00e9rez Boada En esta tesis se han abordado estudios estructura-funci\u00f3n de un nuevo tipo de peroxidasa [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center 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