Abstract: The present disclosure relates to a non-naturally occurring microorganism that includes a gene encoding a MucK transporter protein, where the microorganism is capable of catabolizing terephthalic acid (TPA). In some embodiments of the present disclosure, the gene encoding the MucK transporter protein may contain at least one mutation, relative to a reference gene encoding a reference MucK transporter protein.

Abstract: Disclosed herein are methods and compositions for catalytic glycolysis to deconstruct PET to bis(2-hydroxyethyl) terephthalate (BHET). For BHET conversion to terephthalate and ethylene glycol, we engineer Pseudomonas putida KT2440 with PETase and MHETase enzymes from Ideonella sakaiensis. We further engineer P. putida to convert terephthalate to a performance-advantaged bioproduct, ?-ketoadipic acid, and for improved utilization of ethylene glycol, a byproduct of BHET catabolism. In a bioreactor, we produce 15.1±0.6 g/L of ?-ketoadipic acid (?KA) from BHET at 76±3% molar yield. Lastly, we demonstrate conversion of catalytically depolymerized PET to ?KA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.

Abstract: Disclosed herein are engineered P. putida KT2440 co-expressing PETase and MHETase enzymes that selectively degrades PET into monomers, ethylene glycol and terephthalate (TPA). In another embodiment, disclosed herein are methods for making and using a highly efficient EG metabolizing P. putida KT2440 strain. Given that native P. putida does not have a TPA metabolic pathway, nor the proteins to transport TPA into the cell, the next metabolic engineering challenge for developing synthetic P. putida strain to plastic upcycling was enabling TPA catabolism in P. putida KT2440. TPA transporters and catabolic pathway have been characterized in several microorganisms including Comamonas sp. strain E6 and Rhodococcus jostii RHA1.