Engineered Microorganisms for the Deconstruction of Polymers

Gregg Beckham; Lahiru Niroshan Jayakody Thelhawadigedara; Adam Guss; Thomas Mand; Christopher Johnson; Isabel Pardo Mendoza; Allison Werner

Engineered Microorganisms for the Deconstruction of Polymers

Gregg Beckham (Inventor)
, Lahiru Niroshan Jayakody Thelhawadigedara (Inventor)
, Adam Guss (Inventor)
, Thomas Mand (Inventor)
, Christopher Johnson (Inventor)
, Isabel Pardo Mendoza (Inventor)
, Allison Werner (Inventor)

Research output: Patent

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, ..beta..-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 ..beta..-ketoadipic acid (..beta..KA) from BHET at 76+/-3% molar yield. Lastly, we demonstrate conversion of catalytically depolymerized PET to ..beta..KA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.

Original language	American English
Patent number	12,371,718 B2
Filing date	29/07/25
State	Published - 2025

NLR Publication Number

NREL/PT-2A00-96346

Keywords

exogenous gene addition
genetically modified organism
MHETase
PETase

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Cite this

@misc{6009efc10b1e421390badd6664d7ff68,

title = "Engineered Microorganisms for the Deconstruction of Polymers",

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, ..beta..-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 ..beta..-ketoadipic acid (..beta..KA) from BHET at 76+/-3\% molar yield. Lastly, we demonstrate conversion of catalytically depolymerized PET to ..beta..KA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.",

keywords = "exogenous gene addition, genetically modified organism, MHETase, PETase",

author = "Gregg Beckham and Thelhawadigedara, \{Lahiru Niroshan Jayakody\} and Adam Guss and Thomas Mand and Christopher Johnson and \{Pardo Mendoza\}, Isabel and Allison Werner",

year = "2025",

language = "American English",

publisher = "National Renewable Energy Laboratory (NREL)",

address = "United States",

type = "Patent",

note = "12,371,718 B2",

}

TY - PAT

T1 - Engineered Microorganisms for the Deconstruction of Polymers

AU - Beckham, Gregg

AU - Thelhawadigedara, Lahiru Niroshan Jayakody

AU - Guss, Adam

AU - Mand, Thomas

AU - Johnson, Christopher

AU - Pardo Mendoza, Isabel

AU - Werner, Allison

PY - 2025

Y1 - 2025

N2 - 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, ..beta..-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 ..beta..-ketoadipic acid (..beta..KA) from BHET at 76+/-3% molar yield. Lastly, we demonstrate conversion of catalytically depolymerized PET to ..beta..KA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.

AB - 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, ..beta..-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 ..beta..-ketoadipic acid (..beta..KA) from BHET at 76+/-3% molar yield. Lastly, we demonstrate conversion of catalytically depolymerized PET to ..beta..KA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.

KW - exogenous gene addition

KW - genetically modified organism

KW - MHETase

KW - PETase

M3 - Patent

M1 - 12,371,718 B2

Y2 - 2025/07/29

PB - National Renewable Energy Laboratory (NREL)

ER -

Engineered Microorganisms for the Deconstruction of Polymers

Abstract

NLR Publication Number

Keywords

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