Enhanced synthesis of medium-chain-length poly(3-hydroxyalkanoates) by inactivating the tricarboxylate transport system of Pseudomonas putida KT2440 and process development using waste vegetable oil |
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| Institution: | 1. Biosystems Engineering Laboratory, Center for Bioinformatics and Integrative Biology (CBIB), Faculty of Biological Sciences, Universidad Andres Bello, República 239, 8370146 Santiago, Chile;2. BioNanotechnology and Microbiology Laboratory, Center for Bioinformatics and Integrative Biology (CBIB), Faculty of Biological Sciences, Universidad Andres Bello, República 239, 8370146 Santiago, Chile;3. Departamento de Química Orgánica, Facultad de Química, Pontificia Universidad Católica de Chile, Vicuña Mackenna 4860, Macul, 78204336 Santiago, Chile;1. Department of Biosystems Engineering, University of Manitoba, Winnipeg, Manitoba, R3T 5V6, Canada;2. Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada;3. Scion Research, Te Papa Tipu Innovation Park, 49 Sala Street, Private Bag 3020, Rotorua, 3046, New Zealand;1. School of Biomolecular and Biomedical Science, University College Dublin, Belfield, Dublin 4, Ireland;2. Technology Centre for Biorefining and Bioenergy, Orbsen Building, National University of Ireland, Galway, Ireland;3. Centre for Research Adoptive Nanostructures and Nano Devices, Trinity College, Dublin 2, Ireland;4. Department of Biochemistry, National University of Ireland, Galway, Ireland;5. School of Biochemical and Bioprocessing Engineering, University College Dublin, Belfield, Dublin 4, Ireland;6. School of Physics, Trinity College, Dublin 2, Ireland |
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| Abstract: | The use of waste materials as feedstock for biosynthesis of valuable compounds has been an intensive area of research aiming at diminishing the consumption of non-renewable materials. In this study, P. putida KT2440 was employed as a cell factory for the bioconversion of waste vegetable oil into medium-chain-length Polyhydroxyalkanoates. In the presence of the waste oil this environmental strain is capable of secreting enzymes with lipase activities that enhance the bioavailability of this hydrophobic carbon substrate. It was also found that the oxygen transfer coefficient is directly correlated with high PHA levels in KT2440 cells when metabolizing the waste frying oil. By knocking out the tctA gene, encoding for an enzyme of the tripartite carboxylate transport system, an enhanced intracellular level of mcl-PHA was found in the engineered strain when grown on fatty acids. Batch bioreactors showed that the KT2440 strain produced 1.01 (g?L?1) of PHA whereas the engineered ΔtctA P. putida strain synthesized 1.91 (g?L?1) after 72 h cultivation on 20 (g?L?1) of waste oil, resulting in a nearly 2-fold increment in the PHA volumetric productivity. Taken together, this work contributes to accelerate the pace of development for efficient bioconversion of waste vegetable oils into sustainable biopolymers. |
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| Keywords: | Waste vegetable oil Polyhydroxyalkanoates Tricarboxylate acid transporter Lipase |
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