{"id":113352,"date":"2018-03-11T10:40:46","date_gmt":"2018-03-11T10:40:46","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/caracterizacion-cinetica-y-molecular-de-nicotin-desaminasas-y-aldolasas-mediante-enzimas-acopladas\/"},"modified":"2018-03-11T10:40:46","modified_gmt":"2018-03-11T10:40:46","slug":"caracterizacion-cinetica-y-molecular-de-nicotin-desaminasas-y-aldolasas-mediante-enzimas-acopladas","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/bioquimica-molecular\/caracterizacion-cinetica-y-molecular-de-nicotin-desaminasas-y-aldolasas-mediante-enzimas-acopladas\/","title":{"rendered":"Caracterizacion cinetica y molecular de nicotin desaminasas y aldolasas mediante enzimas acopladas"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Guiomar Sanchez Carron <\/strong><\/h2>\n<p>Palabras clave: \u00e1cido n-acetilneuram\u00ednico aldolasa, enzima acoplada, cin\u00e9tica, ensayo espectrofotom\u00e9trico, lactato deshidroganasa, evoluci\u00f3n dirigida, nicotinaminidasa, glutamato deshidroganasa, nicotinamida mononucleotido desaminasa, modelado de estructuras prot\u00e9icas, filogenia, mutag\u00e9nesis, metabolismo nad.   resumen en espa\u00f1ol: el objetivo de esta tesis fue la b\u00fasqueda de nuevas aldolasas y nicotin desaminasas bacterianas con caracter\u00edsticas mejoradas para la producci\u00f3n de compuestos con inter\u00e9s farmac\u00e9utico y biotecnol\u00f3gico. En primer lugar, se clon\u00f3 y caracteriz\u00f3 una \u00e1cido n-acetilneuram\u00ednico aldolasa de lactobacillus plantarum, que se utiliz\u00f3 como molde para la creaci\u00f3n de librer\u00edas de evoluci\u00f3n dirigida. Se desarrollo un m\u00e9todo de ensayo de alto rendimiento para esta librer\u00eda de evoluci\u00f3n dirigida, utilizando una l-lactato deshidrogenasa de bacillus halodurans como enzima acoplada. 2 nuevas enzimas involucradas en el metabolismo del nad se clonaron y caracterizaron: una nicotinaminidasa y una nicotinamida mononucle\u00f3tido (nmn) desaminasa, ambas obtenidas del extrem\u00f3filo oceanobacillus iheyensis hte831. Se desarrollo tambi\u00e9n un m\u00e9todo de cribado de alto rendimiento para nicotinaminidasas, basado en el uso de una glutamato deshidrogenasa clonada de bacillus halodurans. Adem\u00e1s, se describi\u00f3 por primera vez un m\u00e9todo de medida espectrofotom\u00e9trico para la nmn desaminasa.   resumen en ingl\u00e9s: the main objective was the search for new bacterial aldolases and nicotin deamidases with improved characteristics for the production of compounds with pharmaceutical and biotechnological interest. In first instance, the cloning and characterization of the novel n-acetylneuraminic acid aldolase from lactobacillus plantarum was achieved and it was used as template for directed evolution experiments. A high-throughput screening method for this directed evolution library was developed using a novel l-lactate dehydrogenase from bacillus halodurans as coupling enzyme. Two novel enzymes involved in the nad+ metabolism were also cloned and characterized: a nicotinamidase and a nicotin mononucleotide (nmn) deamidase, both from the extremophile oceanobacillus iheyensis hte831. A high-throughput screening method for nicotinamidases was also developed, based on the use of a cloned glutamate dehygrogenase from bacillus halodurans. Furthermore, a novel spectrophotometric screening method was described for the first time for the enzyme nicotinamide mononucleotide deamidase.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Caracterizacion cinetica y molecular de nicotin desaminasas y aldolasas mediante enzimas acopladas<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Caracterizacion cinetica y molecular de nicotin desaminasas y aldolasas mediante enzimas acopladas <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Guiomar Sanchez Carron <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Murcia<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 26\/10\/2012<\/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>Francisco Garc\u00eda Carmona<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: mercedes Jim\u00e9nez ati\u00e9nzar <\/li>\n<li>Fernando Gil ortiz (vocal)<\/li>\n<li>silvia Montoro Garc\u00eda (vocal)<\/li>\n<li>pilar Zafrilla rentero (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Guiomar Sanchez Carron Palabras clave: \u00e1cido n-acetilneuram\u00ednico aldolasa, enzima acoplada, cin\u00e9tica, ensayo espectrofotom\u00e9trico, lactato deshidroganasa, evoluci\u00f3n dirigida, [&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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