Engineered microbes for the production of biopolymers from ohd-derived organic carbon
AssigneeBoard of Trustees of Southern Illinois University
InventorsLahiru N. Jayakody, Kenneth B. Anderson
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Abstract
This disclosure generally relates to engineered recombinant bacterial cells producing enzymatic compositions for up-cycling plastics, biomass waste, or co-mixtures of plastics and biomass waste. The disclosure also relates to dual enzyme compositions produced by the recombinant bacterial cells for upcycling plastic materials and/or biomass waste, and methods for upcycling plastics, OHD-treated substrates and/or biomass waste.
Claims23
A recombinant bacterial cell comprising a heterologous DNA encoding and expressing at least one heterologous PETase-like enzyme and at least one heterologous MHETase-like enzyme,
- wherein the PETase-like enzyme has a secretion signal peptide linked in frame to an enzymatic activity for degrading Bis(2-hydroxyethyl) terephthalate (BHET) into Mono-(2-hydroxyethyl)terephthalic acid (MHET) and comprises Leaf-Compost Cutinase (LCC) enzyme with SEQ ID NO:
- 5, or a functional variant thereof having at least 85% overall sequence identity to SEQ ID NO:
- 5;
- and
- wherein the MHETase-like enzyme has a secretion signal peptide linked in frame to an enzymatic activity for degrading MHET into ethylene glycol and terephthalic acid and comprises a polypeptide with SEQ ID NO:
- 6, a functional variant thereof having at least 85% overall sequence identity to SEQ ID NO:
- 6, Mle046 enzyme with SEQ ID NO:
- 13, a functional variant thereof having at least 85% overall sequence identity to SEQ ID NO:
- 13, Mle046 mutant enzyme with SEQ ID NO:
- 14, a functional variant thereof having at least 85% overall sequence identity to SEQ ID NO:
- 14, or any combination thereof.
A dual enzyme composition comprising at least one PETase-like enzyme and at least one MHETase-like enzyme,
- wherein the PETase-like enzyme comprises at least amino acids 28-320 of the polypeptide with SEQ ID NO:
- 5 and/or a functional variant therefore having at least 80% overall sequence identity to amino acids 28-320 of SEQ ID NO:
- 5, or any combination thereof, wherein the PETase-like enzyme having an enzymatic activity for degrading bis(2-hydroxyethyl) terephthalate (BHET) to mono-(2-hydroxyethyl)terephthalic acid (MHET);
- and
- wherein the MHETase-like enzyme comprises at least amino acids 18-613 of the polypeptide with SEQ ID NO:
- 6 and/or a functional variant thereof having at least 80% overall sequence identity to amino acids 28-320 of SEQ ID NO:
- 5, or any combination thereof, and/or Mle046 enzyme comprising SEQ ID NO:
- 13 and/or a functional variant therefore having at least 80% overall sequence identity to SEQ ID NO:
- 13, and/or mutant Mle046 enzyme comprising SEQ ID NO:
- 14, or a functional variant thereof having at least 80% overall sequence identity to SEQ ID NO:
- 14, and wherein the MHETase-like enzyme has an enzymatic activity for degrading MHET to terephthalic acid (TPA) and ethylene glycol (EG).
A method for producing cis-cis muconate, the method comprising contacting a substrate comprising terephthalic acid (TPA) with a recombinant bacterial cell containing a mutation in its genome, the mutation eliminating expression of muconate cycloisomerase enzymatic activity with SEQ ID NO:
- 4, wherein the mutation is a deletion and/or insertion of at least one nucleotide or more and wherein the substrate is produced by degradation of biomass waste and/or plastic material.
Dependencies are inferred from the claim text.