THE GOAL OF THE PROPOSED RESEARCH IS TO DEVELOP BIOTECHNOLOGY THAT WILL ENABLE BROADER USE OF SUNFLOWER SEED-DERIVED INGREDIENTS IN HIGH PH BAKING APPLICATIONS. SUNFLOWER SEED-DERIVED INGREDIENTS REPRESENT ALTERNATIVES TO TREE NUT AND LEGUME-BASED PROTEINS, BUTTERS, AND SPREADS, WHICH CAUSE ALLERGIC REACTIONS FOR MANY PEOPLE. WHERE A LIGHT COLOR IS DESIRED IN HIGH PH APPLICATIONS, THE USE OF SUNFLOWER SEED-DERIVED INGREDIENTS (SUNFLOWER SEED BUTTER, FLOUR, AND MEAL), ISHINDERED BY THE HIGH CONCENTRATION OF CHLOROGENIC ACID IN SUNFLOWER SEEDS. CHLOROGENIC ACID REACTS WITH FOOD COMPONENTS SUCH AS AMINO ACIDS AT HIGH TEMPERATURE AND ALKALINE PH, COMMON CONDITIONS ENCOUNTERED IN BAKING. THIS REACTION PRODUCES AN INTENSE GREEN COLOR THAT CAN LIMIT THE APPLICATION OF SUNFLOWER SEED-DERIVED INGREDIENTS IN PRODUCTS WITH AN ALKALINE PH WHERE A WHITE TO BEIGE COLOR IS WANTED. TO DATE, THERE IS NO KNOWN METHOD OF ENTIRELY ELIMINATING THIS GREENING WITHOUT ACIDS OR SIMULTANEOUS REMOVAL OF BENEFICIAL ANTIOXIDANT COMPOUNDS THAT ARE PRESENT IN SUNFLOWER SEED-DERIVED INGREDIENTS. TO OVERCOME THIS PROBLEM, WE PROPOSE A NEW WAY OF REMOVING CHLOROGENIC ACID. WE WILL USE A CHLOROGENIC ACID ESTERASE ENZYME TO BREAK DOWN CHLOROGENIC ACID AND THEREBY ELIMINATE GREENING. THIS APPROACH WILL ENABLE THE CLEAVAGE OF CHLOROGENIC ACID WITHOUT AFFECTING DESIRABLE ANTIOXIDANT COMPOUNDS.THE MAJOR INNOVATION OF THE PROPOSED RESEARCH IS TO ENGINEER CHLOROGENIC ACID ESTERASE WITH HIGH ACTIVITY AND STABILITY FOR USE IN BAKING APPLICATIONS. THE CHLOROGENIC ESTERASE WE WILL USE IS FROM LACTOBACILLUS, A GENUS OF BACTERIA THAT IS ALREADY IN USE IN FOOD MANUFACTURING. WE WILL ENGINEER THE ESTERASE BASED ON STATE-OF-THE-ART COMPUTATIONALLY-AIDED PROTEIN DESIGN STRATEGIES FOR MAXIMAL ACTIVITY AND STABILITY. WE WILL THEN PRODUCE THE ESTERASE IN LARGE QUANTITIES IN THE BACTERIUME. COLI, LYSE THE BACTERIUM AND PURIFY THE ENZYME. AFTER THAT, WE WILL UTILIZE THE PURIFIED ENZYME TO BREAK DOWN CHLOROGENIC ACID IN SUNFLOWER MEAL AND FLOUR. THIS ORIGINAL APPROACH CAN POTENTIALLY GENERATE A MARKETABLE ENZYME FOR THEFOOD INDUSTRY. FURTHERMORE, SINCE ENZYMES ARE BIOLOGICAL MOLECULES, WE EXPECT POSITIVE CONSUMER PERCEPTION OF THE SUNFLOWER SEED-DERIVED INGREDIENTS COLORED. IF SUCCESSFUL, WORK STEMMING FROM THIS PROPOSAL WILL CONVERT SUNFLOWER SEED INGREDIENTS SUCH AS FLOUR OR THE MEAL, WHICH IS CURRENTLY A LOW-VALUE BYPRODUCT OF OILSEED-PRESSING THAT IS MAINLY USED AS ANIMAL FODDER, INTO A DESIRABLE, HIGH-VALUE BAKING PRODUCT.