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Poly[(R)-3-hydroxyalkanoates] (PHAs) are biodegradable polymeric materials synthesized by gram negative bacteria from sugars or fatty acid carbon feedstocks. PHAs are produced by microorganisms when they are grown under nutrient limiting conditions with an excess of carbon sources. These high amounts of carbon feedstocks get converted into PHAs with the purpose of providing the microorganisms with a carbon reserve. The PHAs get packaged into granules that can be harvested by scientists to make a grand variety of products. Among the most innovative uses for PHAs are their biomedical applications that include sutures, drug delivery capsules and tissue engineering of heart valves. These materials do not require photochemical nor mechanical processes for degradation, they get broken down by other microorganisms and enzymes to carbon dioxide and water. Our contribution relates to the role of PP0763 in the metabolic network pertaining to the conversion of fatty acids into PHAs present in the gram negative soil bacterium Pseudomonas putida KT2440. This protein is hypothesized to have 3-hydroxyacyl-CoA ligase activity responsible for the synthesis of the 3-hydroxyacyl-CoA PHA precursor. Here we report our efforts in elucidating the substrate specificities of PP0763 by employing the DTNB colorimetric assay to measure the kinetics of fatty acyl-CoA synthesis in the presence of fatty acids of varying chain lengths and chemical compositions.
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Janer Pagan, Nataniel, "Kinetic Characterization of PP0763: A protein involved in the production of Poly[(R)-3-hydroxyalkanoates] (PHA’s)" (2022). Senior Projects Spring 2022. 48.
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