{"id":15742,"date":"2018-03-09T09:03:02","date_gmt":"2018-03-09T09:03:02","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/estudio-de-la-produccion-heterologa-de-una-lipasa-del-hongo-rhizopus-oryzae-en-la-levadura-metilotrofica-pichia-pastoris\/"},"modified":"2018-03-09T09:03:02","modified_gmt":"2018-03-09T09:03:02","slug":"estudio-de-la-produccion-heterologa-de-una-lipasa-del-hongo-rhizopus-oryzae-en-la-levadura-metilotrofica-pichia-pastoris","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/quimica\/estudio-de-la-produccion-heterologa-de-una-lipasa-del-hongo-rhizopus-oryzae-en-la-levadura-metilotrofica-pichia-pastoris\/","title":{"rendered":"Estudio de la producci\u00f3n heter\u00f3loga de una lipasa del hongo rhizopus oryzae en la levadura metilotr\u00f3fica pichia pastoris"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Alicia Serrano Gonz\u00e1lez <\/strong><\/h2>\n<p>El trabajo recoge los principales aspectos de la producci\u00f3n recombinante de una lipasa del hongo rhizopus oryzae (rol) expresada en la levadura pichia pastoris. El sistema de expresi\u00f3n empleado est\u00e1 basado en la utilizaci\u00f3n del promotor de la alcohol oxidasa (p aox1) de la levadura y, por lo tanto es dependiente de la utilizaci\u00f3n de metanol como substrato inductor. El trabajo recoge una serie de estudios de producci\u00f3n en bioreactor, operando en fed-batch, en los que la productividad del sistema se incrementa hasta en un factor de 11, mediante el estudio de la estrategia de adici\u00f3n del metanol a lo largo de la etapa de producci\u00f3n. Asimismo, estos cultivos se han realizado utilizando un medio de composici\u00f3n sint\u00e9tica, mucho m\u00e1s econ\u00f3mico que el complejo. La optimizaci\u00f3n de la producci\u00f3n tambi\u00e9n se ha abordado mediante la construcci\u00f3n de cepas multicopia para el gen rol, estudiando igualmente, el efecto de la expresi\u00f3n recombinante de la lipasa sobre la cepa hospedadora.  por otro lado, se han construido vectores de expresi\u00f3n basados en la utilizaci\u00f3n de promotores independientes de la utilizaci\u00f3n del metanol. Se han obtenido cepas de p.Pastoris que expresan la rol de forma constitutiva y mediante inducci\u00f3n por fuente de nitr\u00f3geno.  el trabajo recoge en \u00faltimo lugar un protocolo para la recuperaci\u00f3n de la rol recombinante desde el caldo de cultivo hasta la obtenci\u00f3n de un extracto liofilizado de lipasa rol. La purificaci\u00f3n de la lipasa se ha llevado a acabo a trav\u00e9s de la combinaci\u00f3n de dos pasos de cromatograf\u00eda, de intercambio cati\u00f3nico e interacci\u00f3n hidrof\u00f3bica. El grado de pureza obtenido es el adecuado para utilizaci\u00f3n de la enzima como biocatalizador en reacciones de qu\u00edmica fina y para la obtenci\u00f3n de anticuerpos contra la misma.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Estudio de la producci\u00f3n heter\u00f3loga de una lipasa del hongo rhizopus oryzae en la levadura metilotr\u00f3fica pichia pastoris<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Estudio de la producci\u00f3n heter\u00f3loga de una lipasa del hongo rhizopus oryzae en la levadura metilotr\u00f3fica pichia pastoris <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Alicia Serrano Gonz\u00e1lez <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Aut\u00f3noma de barcelona<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 22\/02\/2002<\/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 Valero Barranco<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: josep Lopez santin <\/li>\n<li>\u00e1ngel Dom\u00ednguez olavarri (vocal)<\/li>\n<li>Mar\u00eda Molina mart\u00edn (vocal)<\/li>\n<li>Antonio Villaverde corrales (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Alicia Serrano Gonz\u00e1lez El trabajo recoge los principales aspectos de la producci\u00f3n recombinante de una lipasa del [&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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