Progress Toward the Biochemical Characterization of a Protein Involved in the Production of Microbial Plastics: Asymmetric Synthesis of 3-Hydroxy Fatty Acids

Poly(3-hydroxyalkanoates) (PHAs) are biodegradable polymeric materials that are of interest due to their ability to serve as green alternatives to the less environmentally friendly petroleum-based plastics that populate our modern world. PHAs are produced by various microorganisms from sugar and fatty acid feedstocks, and they are degraded by microorganisms as well. The metabolic fundamentals of the production of PHAs can be investigated through studying the behavior of the protein PP_0763 from the soil bacterium Pseudomonas putida KT2440, which we believe to be involved in the metabolic redirection of intracellular fatty acids toward PHA biosynthesis. PP_0763 has been successfully expressed and purified, and insight can now be gained into the substrate specificity of this protein via biochemical assay. In order to move forward with assay design, in this study we demonstrate our progress towards the synthesis of a panel of enantiopure 3-hydroxy fatty acids via an asymmetric aldol strategy. Once complete, our assay will test the protein’s affinity for 3-hydroxy fatty acids of varying hydrocarbon chain lengths, leading to clarification of the role the protein plays in PHA biosynthesis. Senior Project submitted to The Division of Science, Mathematics and Computing of Bard College