CHEMICAL BIOSYNTHESIS IS A SIGNIFICANTLY GROWING SECTOR IN INDUSTRIAL BIOTECHNOLOGY. OVER DECADES OF RESEARCH, REMARKABLE ADVANCEMENTS IN METABOLIC ENGINEERING HAVE PROVIDED THE TECHNICAL ABILITY TO CONSTRUCT MICROBIAL CELL FACTORIES BASED ON CONVENTIONAL WORKHORSE MICROORGANISMS SUCH AS SACCHAROMYCES CEREVISIAE. ALTHOUGH THESE APPROACHES HAVE RAPIDLY AND EFFICIENTLY ENABLED THE DEVELOPMENT OF DESIGNER MICROORGANISMS, THE INHERENT LIMITATIONS OF CONVENTIONAL STRAINS OFTEN RESULT IN UNCOMPETITIVE BIOPROCESSING DUE TO LOW PRODUCTIVITY IN HYDROLYSATES AND HIGH PRODUCTION COSTS. WE CURRENTLY LACK NEW PLATFORMS THAT OVERCOME CHALLENGES CAUSED BY INHERENT LIMITATIONS IN MODEL HOST STRAINS.ENGINEERING NON-CONVENTIONAL MICROBES HAS RECENTLY EMERGED AS A POTENTIAL STRATEGY FOR ECLIPSING THESE LIMITATIONS. NON-CONVENTIONAL STRAINS' NATIVE STRESS TOLERANCE HAS CREATED A NEW OVERPRODUCTION CHASSIS FOR BIOSYNTHESIS. THIS PROJECT WILL SEEK TO ESTABLISH A RATIONAL FRAMEWORK FOR ENGINEERING OF NON-MODEL MICROBIAL PLATFORMS TO EXPAND UTILIZATION OF RENEWABLE CARBON SOURCES. CONVERSION OF AGRICULTURAL BIOMASS TO VALUE-ADDED PRODUCTS USING MICROBIAL FERMENTATION IS AN ATTRACTIVE OPTION TO SUBSTITUTE PETROLEUM-BASED PRODUCTION ECONOMICALLY AND SUSTAINABLY. WITH RISING ENERGY DEMAND AND ENVIRONMENTAL POLLUTION, THE DEVELOPMENT OF BIOSYSTEMS AND MICROBIAL CELL FACTORIES AMENABLE TO GENOME ENGINEERING WILL POSITION THESE ORGANISMS AS NEXT-GENERATION PLATFORMS FOR PRODUCING BIO-BASED PRODUCTS FROM RENEWABLE FEEDSTOCKS. WE WILL INTRODUCE HETEROLOGOUS METABOLIC PATHWAYS FOR NON-CONVENTIONAL CARBON SOURCES IN NON-MODEL YEAST ISSATCHENKIAORIENTALIS. I ORIENTALIS IS KNOWN FOR ITS ABILITY TO TOLERATE A VARIETY OF STRESSES, INCLUDING LOW PH AND HIGH TEMPERATURE. BASED ON THE RESULTS FROM I. ORIENTALIS, THIS PROJECT CAN EXPAND HOST STRAINS INTO OTHER NON-MODEL MICROORGANISMS. SUCCESSFUL COMPLETION OF OUR PROPOSAL WILL ESTABLISH A FIRST-OF-ITS-KIND, VERSATILE NON-MODEL YEAST PLATFORM FOR NEXT-GENERATION CHEMICAL BIOSYNTHESIS. FINALLY, THE RECONFIGURED CENTRAL METABOLISM IN STRESS-TOLERANT YEAST WILL SUBSTANTIALLY EXPAND THE VERSATILITY OF RENEWABLE RESOURCE SYSTEMS FOR INNOVATIVE BIOTECHNOLOGICAL APPLICATIONS.