Alliance for Sustainable Energy, LLC

Ut-battelle, Inc.

Other

University of Illinois

University of Georgia Research Foundation, Inc.

Triad National Security, LLC

Montana State University

University of Portsmouth

Christopher W Johnson

Gregg Tyler Beckham

Derek Richard Vardon

Mary Ann Franden

Isabel Pardo Mendoza

Allison Jean Zimont Werner

Jeffrey G Linger

Adam Michael Guss

Thelhawadigedara Lahiru Niroshan Jayakody

Ellen Lee Neidle

Thomas David Mand

Mark Andrew Eiteman

Michael T Guarnieri

Eric M Karp

Philip Thomas Pienkos

Ramesh K Jha

Daniel A Jacobson

Joshua R Elmore

Taraka Dale

Alexander William Meyers

Jennifer DuBois

Melodie M Machovina

Simon James Bradshaw Mallinson

Sandra Fabienne Notonier

Adam Michael Feist

Elsayed Tharwat Tolba Mohamed

Erica Teixeira Prates

John E McGeehan

Davinia Salvachúa Rodríguez

Peter Corbin St John

Timothy J Strathmann

Melissa Tumen-Velasquez

Niju Narayanan

Alaa Ashraf Mahmoud Ahmed

George Lee Peabody, V

Gayle Joann Bentley

Davinia Salvachua Rodriguez

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.

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.

Mutant transporters for bacterial uptake of terephthalic acid

The present disclosure relates to a genetically modified microbial cell that includes a first genetic modification resulting in the expression of an exogenous vanillate demethylase, such that the microbial cell is capable of metabolizing an S-lignin decomposition product and producing 2-pyrone-4,6-dicarboxylate (PDC).

Conversion of S-lignin compounds to useful intermediates

Disclosed herein are the genetically modifiedwith improved tolerance to hydroxycinnamic acids.

Microorganisms engineered for muconate production

The present disclosure relates to a non-naturally occurring microorganism that includes an endogenous genetic deletion that eliminates the expression of at least a pyruvate kinase, where the genetically modified prokaryotic microorganism is capable of producing 3-deoxy-D-arabino-heptulosonate-7-phosphate.

Engineered microorganisms for the production of intermediates and final products

An aspect of the present disclosure is a microbial cell that includes a genetic modification resulting in the expression of a deficient form of an endogenous dioxygenase, and a gene encoding an exogenous dioxygenase and a promoter sequence, where the endogenous dioxygenase includes PcaH and PcaG, the exogenous dioxygenase includes LigA and LigB, the microbial cell is capable of growth utilizing at least one of a cellulose decomposition molecule or a lignin decomposition molecule, and the microbial cell is capable of producing 2-hydroxy-2H-pyran-4,6-dicarboxylic acid.

Conversion of biomass to useful intermediates

Disclosed herein are compositions of non-naturally occurring enzymes to enable microbial syringol utilization with GcoAB.

Microbial syringol utilization

The present disclosure relates to a genetically modified microbial cell that includes a genetic modification resulting in the expression of a vanillate demethylase, where the microbial cell is capable of metabolizing at least one S-lignin decomposition molecule including at least one of syringate and/or 3-O-methyl gallate, and the genetically modified microbial cell is capable of producing gallate. In some embodiments of the present disclosure, the vanillate demethylase may include VanAB.

Disclosed herein are enzymes and organisms useful for the dealkylation of products derived from lignin depolymerization, including the conversion of guaiacol or guaethol to catechol or the conversion of anisole to phenol. Methods of converting guaiacol or guaethol to catechol or anisole to phenol using enzymes or organisms expressing the same are also disclosed.

Enzymes and methods for dealkylation of substrates

This disclosure relates to compositions and methods for converting biomass to various chemical intermediates and final products including fuels. Aspects include the depolymerization of lignin, cellulose, and hemicellulose to a wide slate of depolymerization compounds that can be subsequently metabolized by genetically modified bacterium, and converted to cis,cis-muconic acid. Other aspects include the use of monometallic catalysts for converting the cis,cis-muconic acid to commodity chemicals and fuels, for example adipic acid and/or nylon.

Biomass conversion to fuels and chemicals

Disclosed herein are methods for the production of fuels, chemicals, or materials from biomass-derived lignin, cellulose, and hemicellulose. Also provided are methods for the production of polyhydroxyalkanoates from lignocellulosic biomass and the subsequent thermal and catalytic conversion of polyhydroxyalkanoates to hydrocarbons. In addition, methods for the production of ethanol from the fermentation of polysaccharides are described.

Lignin conversion to fuels, chemicals and materials

Disclosed herein are enzymes useful for the dealkylation of aromatic substrates, including the conversion of guaiacol or guaethol to catechol. Methods of converting aromatic substrates found in lignin-based feedstocks such as pyrolysis oil into products such as catechol are also disclosed. Also presented herein are methods for rapidly evolving and optimizing genetic regions.

Chimeric enzymes for conversion of lignin-derived chemicals

Growing community of inventors

Denver, CO, United States of America

Christopher W Johnson