Ut-battelle, Inc.

Alliance for Sustainable Energy, LLC

Thomas David Mand

Gregg Tyler Beckham

Christopher W Johnson

Thelhawadigedara Lahiru Niroshan Jayakody

Isabel Pardo Mendoza

Adam Michael Guss

Allison Jean Zimont Werner

Disclosed herein are engineeredKT2440 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 metabolizingKT2440 strain. Given that nativedoes not have a TPA metabolic pathway, nor the proteins to transport TPA into the cell, the next metabolic engineering challenge for developing syntheticstrain to plastic upcycling was enabling TPA catabolisminKT2440. TPA transporters and catabolic pathway have been characterized in several microorganisms includingsp. strain E6 andRHA1.

Engineered microorganisms for the deconstruction of polymers

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 engineerKT2440 with PETase and MHETase enzymes from. We further engineerto convert terephthalate to a performance-advantaged bioproduct, β-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 β-ketoadipic acid (βKA) from BHET at 76±3% molar yield. Lastly, we demonstrate conversion of catalytically depolymerized PET to βKA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.

Growing community of inventors

Oak Ridge, TN, United States of America

Thomas David Mand