OBJECTIVES: THE OBJECTIVE OF THIS PROJECT IS TO DEVELOP A CYANOBACTERIAL PLATFORM THAT CONVERTS CO2 TO LIGHTWEIGHT HIGH-PERFORMANCE MATERIALS FOR NASA APPLICATIONS. FROM A SPACE EXPLORATION PERSPECTIVE CYANOBACTERIA SUCH AS SYNECHOCYSTIS SP. PCC 6803 HAVE THE DESIRABLE CAPACITY TO CAPTURE AND STORE ENERGY THROUGH PHOTOSYNTHESIS (CONSUMING CO2 AND REGENERATING O2) AND TO THRIVE UNDER EXTREME CONDITIONS INCLUDING CO2-RICH ATMOSPHERES. FURTHERMORE SYNECHOCYSTIS HAS GREAT METABOLIC POTENTIAL AND HAS BEEN ENGINEERED TO PRODUCE A VARIETY OF USEFUL CHEMICALS FROM CO2 WITH HIGH EFFICIENCIES. MEANWHILE NATURE HAS EVOLVED NUMEROUS PROTEINBASED MATERIALS WITH INTRICATE HIERARCHICAL/MULTI-SCALE STRUCTURES SUCH AS SPIDER SILK RESILIN AND ELASTIN. THESE PROTEIN POLYMERS HAVE REMARKABLE PHYSICAL AND MECHANICAL PROPERTIES INCLUDING EXTREMELY LIGHTWEIGHT HIGH-STRENGTH (EVEN EXCEEDING THAT OF STEEL) HIGH ELASTICITY AND HIGH FATIGUE LIFETIME. THIS PROJECT AIMS TO HARNESS AND COMBINE THE METABOLIC CAPACITY OF CYANOBACTERIA WITH BIOLOGICAL MACHINERY FOR HIERARCHICAL MATERIAL SYNTHESIS TO PROVIDE A POWERFUL SYNTHETIC BIOLOGY PLATFORM FOR PRODUCING LIGHTWEIGHT HIGHSTRENGTH MATERIALS FROM CO2. HERE WE FOCUS ON PROTEIN-BASED MATERIALS BECAUSE OF THEIR VERSATILE FUNCTION AND THE EASE OF DOWNSTREAM PROCESSING AFTER SYNTHESIS. MANUFACTURED PROTEIN FIBERS CAN BE EASILY HARVESTED AND SPUN USING LIMITED RESOURCES WITHOUT COMPLICATED PURIFICATION STEPS TO REMOVE UNDESIRABLE BYPRODUCTS AS NEEDED FOR THE PRODUCTION OF FOOD AND THERAPEUTICS. METHODS: CURRENT BIOSYNTHESIS OF PROTEIN-BASED MATERIALS IS LIMITED TO THE RECOMBINANT PRODUCTION OF A FEW SPIDER SILK AND ELASTOMERIC PROTEINS WHICH HAVE SO FAR FAILED TO REPLICATE THE HIGH MECHANICAL PERFORMANCE OF THEIR NATURAL COUNTERPARTS. THESE FAILURES ARE DUE LARGELY TO THE INABILITY TO SYNTHESIZE ULTRAHIGH MOLECULAR WEIGHT PROTEINS (UHMWS) WITH CONTROLLED ORDERS OF REPEATING SUBUNITS. WE PROPOSE TO SOLVE THESE PROBLEMS BY DEVELOPING A UNIQUE PROTEIN ASSEMBLY STRATEGY. CYANOBACTERIA WILL BE ENGINEERED TO SYNTHESIZE AND SECRET FUNCTIONALIZED PROTEIN MONOMERS. THE SECRETED MONOMERS WILL THEN SELF-LIGATE IN DEFINED ORDERS TO FORM UHMW POLYMERS AND COPOLYMERS WHICH WILL PRECIPITATE FROM THE MEDIA AND CAN BE EASILY HARVESTED FOR DOWNSTREAM PROCESSING. NASA INTERESTS: THE PROPOSED PROJECT WILL PROVIDE A CYANOBACTERIAL PLATFORM FOR THE FACILE PRODUCTION OF LIGHTWEIGHT HIGH-STRENGTH MATERIALS FROM CO2. THE PROPOSED SYNTHETIC BIOLOGY SYSTEM CAN BE USED DURING SPACE TRAVEL TO REGENERATE O2 AND CONSUME CO2 AND AT EXTRATERRESTRIAL NASA SITES TO SUSTAINABLY MANUFACTURE MATERIALS FOR VARIOUS NASA APPLICATIONS SUCH AS LIGHTWEIGHT HIGHSTRENGTH ROPES AND PARACHUTES ENERGY-ABSORBING SEALS UV-PROTECTIVE COATINGS AND DURABLE EQUIPMENT. UNLIKE THE PRODUCTION OF FOOD NUTRACEUTICALS AND THERAPEUTICS FROM ENGINEERED ORGANISM MATERIAL MANUFACTURING IS UNRESTRICTED BY HEALTH CONCERNS RELATED TO HUMAN CONSUMPTION. MOREOVER THE PROPOSED TECHNOLOGY WILL PAVE THE ROAD FOR THE MANUFACTURING OF RENEWABLE MATERIALS BRINGING TRANSFORMATIVE CHANGES TO THE CURRENT MATERIAL INDUSTRY.