RECOMBINANT EXPRESSION OF SPIDER SILK PROTEIN IN PICHIA PASTORIS

THIS PHASE I PROGRAM IS DIRECTED TOWARD THE DEVELOPMENT OF AN EXPRESSION SYSTEM FOR THE HIGH LEVEL SYNTHESIS OF RECOMBINANT FIBROUS PROTEINS IN THE METHYLOTROPHIC YEAST, PICHIA PASTORIS. THE ULTIMATE OBJECTIVE IS A PRODUCTION SYSTEM WHICH PROVIDES HIGH LEVELS OF A RECOMBINANT, SPIDER SILK PROTEIN THAT MAY BE READILY ISOLATED WITH MINIMAL COST, AND WHICH MAY BE SCALED UP WITH MAXIMUM YIELDS OF PRODUCT. DURING THIS DEVELOPMENT PROGRAM, OUR RESEARCH WILL FOCUS ON FOUR PRIMARY AREAS: 1) THE STABILITY OF THE HETEROLOGOUS SPIDER SILK GENE IN THE PICHIA PASTORIS HOST, 2) THE LEVELS OF EXPRESSION FOR SECRETED AND CYTOPLASMIC MODES OF EXPRESSION, 3) THE YIELDS OF PRODUCT FROM ONE-LITER FERMENTORS, AND 4) THE STABILITY AND STRUCTURE OF THE RECOMBINANT PROTEIN.