Optimització del sistema de producció de proteïnes heteròlogues en \i Pichia pastoris\i0 : aplicació a la producció de lipasses i proteïnes terapèutiques

The global objective of the project is the optimisation of the production process of heterologous proteins in \i pichia pastoris\i0 system. \i Rhizopus oryzae\i0 lipase (ROL) is going to be used as model protein, wich has a wide range of applications in fine chemicals. The experience acquire and the developed methodology in ROL will be applied to the production of significant amount of proteins with fine chemicals applications and with know or potential therapeutic activity. In particular the production of the isoenzyme lipase (Lip2) from \i Candida rugosa\i0 and a model therapeutic protein human chorionic gonadotropin (hCG), a heterodimeric glycoprotein hormone used in treatments with infertile patients. Both heterologous proteins are going to be compared with the nativeproteins or produced in other expression systems to validate the capacity of \i Pichia pastoris\i0 expression system from the point of view of biological activity of produced heterologous proteins. To achieve this global objective the project deals with genetic, physiology and metabolism aspects of \i Pichia pastoris\i0 system, culture broth selection, optimisation of fermetation process, downstream and purification, monitoring, mathematical modelling, control and scale-up. Thus, the main goal of the project is the development of a robust battery of tools and methodologies to optimise the production of heterologous proteins in the non-conventional yeast \i Pichia pastoris\i0 . The project is focussed on a multidisciplinary vision to integrate biological and biochemical engineering research and technologies in the fields of health and high value added chemical to achieve the highest productivity and reproductibility of products and processes.