US 12,460,176 B2
Engineered microorganisms for the deconstruction of polymers
Gregg Tyler Beckham, Golden, CO (US); Thelhawadigedara Lahiru Niroshan Jayakody, Wheat Ridge, CO (US); Adam Michael Guss, Oak Ridge, TN (US); Thomas David Mand, Oak Ridge, TN (US); Christopher W. Johnson, Denver, CO (US); and Isabel Pardo Mendoza, Dos Hermanas (ES)
Assigned to Alliance for Sustainable Energy, LLC, Golden, CO (US); and UT-Battelle, LLC, Oak Ridge, TN (US)
Appl. No. 17/055,626
Filed by Alliance for Sustainable Energy, LLC, Golden, CO (US); and UT-Battelle, LLC, Oak Ridge, TN (US)
PCT Filed May 15, 2019, PCT No. PCT/US2019/032480
§ 371(c)(1), (2) Date Nov. 16, 2020,
PCT Pub. No. WO2019/222396, PCT Pub. Date Nov. 21, 2019.
Claims priority of provisional application 62/671,477, filed on May 15, 2018.
Prior Publication US 2021/0180007 A1, Jun. 17, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/52 (2006.01); C12N 1/20 (2006.01); C12N 9/18 (2006.01)
CPC C12N 1/20 (2013.01) [C12N 9/18 (2013.01); C12Y 301/01 (2013.01)] 13 Claims
 
1. A genetically modified Pseudomonas organism comprising at least two exogenous gene additions, wherein:
a first exogenous gene addition comprises a gene with a sequence that is at least 90% identical to SEQ ID NO: 1 and a second exogenous gene addition comprises a gene with a sequence that is at least 90% identical to SEQ ID NO: 2 and wherein the first exogenous gene encodes for a functional PETase comprising a secretion signal peptide and wherein the second exogenous gene encodes for a functional MHETase comprising a secretion signal peptide and wherein the exogenous genes are incorporated into the genome of the genetically modified Pseudomonas; and wherein the genetically modified Pseudomonas organism metabolizes poly (ethylene terephthalate) (PET) to produce PET deconstruction products selected from the group consisting of bis(2-Hydroxyethyl) terephthalate, mono-(2-hydroxyethyl) terephthalate, terephthalate, ethylene glycol, β-ketoadipate, and muconate; and wherein the genetically modified Pseudomonas converts bis(2-hydroxyethyl) terephthalate to terephthalate at a rate that is at least three times the rate of a naturally occurring Pseudomonas.