THE OVERALL GOAL OF THIS PROJECT IS TO FURTHER DEVELOP THE STRATEGY OF ENGINEERING THE MEMBRANE OF MICROBIAL CELL FACTORIES TO IMPROVE PRODUCTION OF BIORENEWABLE FUELS AND CHEMICALS. INHIBITION OF THE MICROBIAL BIOCATALYST OFTEN LIMITS ACHIEVEMENT OF ECONOMICALLY VIABLE TITERS, RATE AND YIELDS. OFTEN, THIS TOXICITY IS ATTRIBUTED TO DAMAGE OF THE CELL MEMBRANE. IF A REACTION VESSEL IS CORRODED BY ITS CONTENTS, A TYPICAL STRATEGY WOULD BE TO CHANGE THE COMPOSITION OF THE VESSEL. HERE, WE AIM TO CHANGE THE COMPOSITION OF THE MICROBIAL CELL MEMBRANE.THE JARBOE GROUP HAS PREVIOUSLY DEMONSTRATED THE FEASIBILITY OF MEMBRANE ENGINEERING FOR IMPROVING TOLERANCE AND PRODUCTION OF FATTY ACIDS AND STYRENE, AS WELL AS TOLERANCE OF THE BIOMASS-DERIVED INHIBITORS FURFURAL AND ACETATE. THIS ENGINEERING FOCUSES ON THREE ASPECTS OF THE MICROBIAL MEMBRANE: THE LIPID TAILS OF THE PHOSPHOLIPIDS, THE HEAD GROUPS OF THE PHOSPHOLIPIDS; AND THE MEMBRANE-ASSOCIATED PROTEINS AND SUGARS.THE SUPPORTING OBJECTIVESOF THIS WORK ARE AS FOLLOWS:1. CHARACTERIZATION AND HIGH-THROUGHPUT SCREENING OF GENETIC CHANGES THAT IMPACT MEMBRANE COMPOSITION AND PROPERTIES.2. COMBINATION OF MEMBRANE ENGINEERING STRATEGIES.3. TUNING OF MEMBRANE ENGINEERING STRATEGIES TO MEMBRANE DAMAGE.THIS PROJECT IS RELEVANT TO THE USDA AFRI PROGRAM IN THAT IT WILL INCREASE THE ECONOMIC VIABILITY OF PLANT PRODUCTION SYSTEMS BY INCREASING THE UTILIZATION OF BIOMASS FOR THE PRODUCTION OF FUELS AND CHEMICALS. THIS PROJECT IS RELEVANT TO THE "BIOPROCESSING AND BIOENGINEERING" PRIORITY OF THE "AGRICULTURE SYSTEMS AND TECHNOLOGY" PROGRAM AREA IN THAT IT WILL IMPROVE THE PRODUCTION EFFICIENCY OF BIO-BASED PRODUCTS.