Modulation of carbon flux through the meg and c3 pathways for the improved production of monoethylene glycol and c3 compounds

A recombinant microbe capable of coproducing MEG and one or more C3 compounds, wherein the microbe comprises one or more of the following

• (i) a disruption of one or more exogenous polynucleotide sequences encoding a phosphate acetyltransferase,
• (ii) a disruption of one or more endogenous polynucleotide sequences encoding an acetate kinase,
• (iii) a disruption of one or more endogenous polynucleotide sequences encoding a pyruvate oxidase,
• (iv) a disruption of one or more endogenous polynucleotide sequences encoding an ArcA regulator,
• (v) a disruption of one or more endogenous polynucleotide sequences encoding a lysine acetyltransferase,
• (vi) one or more overexpressed endogenous polynucleotide sequences encoding a CobB regulator, and
• (vii) one or more overexpressed endogenous polynucleotide sequences encoding an acetyl-CoA synthetase;
• wherein the MEG and/or the one or more C3 compounds are produced at a faster titer, rate or exhibit an increased yield;
• as compared to a microbe lacking the disruption and/or the overexpression of the endogenous polynucleotides of any one or more of i-vii.

A recombinant microbe capable of coproducing MEG and one or more C3 compounds, wherein the microbe comprises one or more of the following

• (i) one or more exogenous polynucleotide sequences encoding a xylose dehydrogenase,
• (i) one or more exogenous polynucleotide sequences encoding a xylonolactonase,
• (iii) one or more exogenous polynucleotide sequences encoding a xylonate dehydratase,
• (iv) one or more exogenous polynucleotide sequences encoding a 3-deoxy-D-glycerol pentanone sugar acid aldolase,
• (v) one or more overexpressed endogenous polynucleotide sequences encoding a xylonate dehydratase,
• (vi) one or more overexpressed endogenous polynucleotide sequences encoding a 3-deoxy-D-glycerol pentanone sugar acid aldolase, and
• (vii) one or more overexpressed endogenous polynucleotide sequences encoding a glycoaldehyde reductase;
• wherein the MEG and/or the one or more C3 compounds are produced at a faster titer, rate or exhibit an increased yield;
• as compared to a microbe lacking the exogenous introduced enzymes and/or the overexpression of the endogenous enzymes of any one or more of i-vii.

A recombinant microbe capable of coproducing MEG and one or more C3 compounds, wherein the microbe comprises:

• (i) one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and/or
• (ii) one or more polynucleotide sequences encoding
• hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase;
• wherein the MEG and/or the one or more C3 compounds are produced at a faster titer, rate or exhibit an increased yield;
• as compared to a microbe not having been introduced an acetoacetyl CoA, hydroxymethylglutaryl-CoA synthase, or hydroxymethylglutaryl-CoA lyase.

A method of making a recombinant microbe capable of coproducing MEG and one or more C3 compounds by:

• modifying a microbe coproducing MEG and one or more C3 compounds by:
• disrupting one or more nucleic acid sequences encoding methylglyoxal synthase(mgsA), and/or
• disrupting one or more nucleic acid sequences encoding glyoxylate carboligase (gcl);
• wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe lacking a disruption of one or more nucleic acid sequences encoding methylglyoxal synthases and/or glyoxylate carboligases;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by performing one or more of the following:
• disrupting one or more exogenous polynucleotide sequences encoding a phosphate acetyltransferase, disrupting one or more endogenous polynucleotide sequences encoding an acetate kinase, disrupting one or more endogenous polynucleotide sequences encoding a pyruvate oxidase, disrupting one or more endogenous polynucleotide sequences encoding an ArcA regulator, disrupting one or more endogenous polynucleotide sequences encoding a lysine acetyltransferase, overexpressing one or more endogenous polynucleotide sequences encoding a CobB regulator, and overexpressing one or more endogenous polynucleotide sequences encoding an acetyl-CoA synthetase;
• wherein the MEG and/or the one or more C3 compounds are produced at a faster titer, rate or exhibit an increased yield;
• as compared to a microbe lacking the disruption and/or the overexpression of the endogenous polynucleotides of any one or more of the above;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by performing one or more of the following:
• introducing one or more exogenous polynucleotide sequences encoding a xylose dehydrogenase, introducing one or more exogenous polynucleotide sequences encoding a xylonolactonase, introducing one or more exogenous polynucleotide sequences encoding a xylonate dehydratase, introducing one or more exogenous polynucleotide sequences encoding a 3-deoxy-D-glycerol pentanone sugar acid aldolase, overexpressing one or more endogenous polynucleotide sequences encoding a xylonate dehydratase, overexpressing one or more endogenous polynucleotide sequences encoding a 3-deoxy-D-glycerol pentanone sugar acid aldolase, and overexpressing one or more endogenous polynucleotide sequences encoding a glycoaldehyde reductase;
• wherein the MEG and/or the one or more C3 compounds are produced at a faster titer, rate or exhibit an increased yield;
• as compared to a microbe lacking the exogenous introduced enzymes and/or the overexpression of the endogenous enzymes of any one or more of the above;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by:
• introducing one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and/or introducing one or more polynucleotide sequences encoding hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase;
• wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe not having been introduced an acetoacetyl CoA, hydroxymethylglutaryl-CoA synthase, or hydroxymethylglutaryl-CoA lyase;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by performing one or more of the following:
• disrupting one or more nucleic acid sequences encoding methylglyoxal synthase (mgsA), disrupting one or more nucleic acid sequences encoding glyoxylate carboligase (gcl), disrupting one or more exogenous polynucleotide sequences encoding a phosphate acetyltransferase, disrupting one or more endogenous polynucleotide sequences encoding an acetate kinase, disrupting one or more endogenous polynucleotide sequences encoding a pyruvate oxidase, disrupting one or more endogenous polynucleotide sequences encoding an ArcA regulator, disrupting one or more endogenous polynucleotide sequences encoding a lysine acetyltransferase, overexpressing one or more endogenous polynucleotide sequences encoding a CobB regulator, overexpressing one or more endogenous polynucleotide sequences encoding an acetyl-CoA synthetase, introducing one or more exogenous polynucleotide sequences encoding a xylose dehydrogenase, introducing one or more exogenous polynucleotide sequences encoding a xylonolactonase, introducing one or more exogenous polynucleotide sequences encoding a xylonate dehydratase, introducing one or more exogenous polynucleotide sequences encoding a 3-deoxy-D-glycerol pentanone sugar acid aldolase, overexpressing one or more endogenous polynucleotide sequences encoding a xylonate dehydratase, overexpressing one or more endogenous polynucleotide sequences encoding a 3-deoxy-D-glycerol pentanone sugar acid aldolase, overexpressing one or more endogenous polynucleotide sequences encoding a glycoaldehyde reductase, introducing one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and introducing one or more polynucleotide sequences encoding hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase;
• wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe lacking the modification;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by:
• disrupting one or more nucleic acid sequences encoding methylglyoxal synthase (mgsA), and disrupting one or more nucleic acid sequences encoding glyoxylate carboligase (gcl) and/or;
• disrupting one or more exogenous polynucleotide sequences encoding a phosphate acetyltransferase, and/or disrupting one or more endogenous polynucleotide sequences encoding an acetate kinase, and/or introducing one or more exogenous polynucleotide sequences encoding a xylonolactonase, and/or introducing one or more exogenous polynucleotide or sequences or overexpressing an endogenous polynucleotide or sequences encoding a xylonate dehydratase, and/or introducing one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and/or introducing one or more polynucleotide sequences encoding hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase;
• wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe lacking a disruption of one or more nucleic acid sequences encoding methylglyoxal synthases and any of the modifications above;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by:
• disrupting one or more nucleic acid sequences encoding glyoxylate carboligase (gcl);
• and disrupting one or more exogenous polynucleotide sequences encoding a phosphate acetyltransferase, and/or disrupting one or more endogenous polynucleotide sequences encoding an acetate kinase, and/or introducing one or more exogenous polynucleotide sequences encoding a xylonolactonase, and/or introducing one or more exogenous polynucleotide or sequences or overexpressing an endogenous polynucleotide or sequences encoding a xylonate dehydratase, and/or introducing one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and/or introducing one or more polynucleotide sequences encoding hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase;
• wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe lacking a disruption of one or more nucleic acid sequences encoding glyoxylate carboligase (gcl) and any of the modifications above;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by:
• disrupting one or more exogenous polynucleotide sequences encoding a phosphate acetyltransferase and disrupting one or more endogenous polynucleotide sequences encoding an acetate kinase, and/or introducing one or more exogenous polynucleotide sequences encoding a xylonolactonase, and/or introducing one or more exogenous polynucleotide or sequences or overexpressing an endogenous polynucleotide or sequences encoding a xylonate dehydratase, and/or introducing one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and/or introducing one or more polynucleotide sequences encoding hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase, and/or;
• disrupting one or more endogenous polynucleotide sequences encoding an ArcA regulator, and/or, disrupting one or more endogenous polynucleotide sequences encoding a lysine acetyltransferase, and/or overexpressing one or more endogenous polynucleotide sequences encoding an acetyl-CoA synthetase;
• wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe lacking a disruption of one or more nucleic acid sequences encoding a phosphate acetyltransferase and any of the modifications above;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by:
• disrupting one or more endogenous polynucleotide sequences encoding an acetate kinase and introducing one or more exogenous polynucleotide sequences encoding a xylonolactonase, and/or introducing one or more exogenous polynucleotide or sequences or overexpressing an endogenous polynucleotide or sequences encoding a xylonate dehydratase, and/or introducing one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and/or introducing one or more polynucleotide sequences encoding hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase;
• wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe lacking a disruption of one or more nucleic acid sequences encoding acetate kinase and any of the modifications above;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by:
• introducing one or more exogenous polynucleotide sequences encoding a xylonolactonase and introducing one or more exogenous polynucleotide or sequences or overexpressing an endogenous polynucleotide or sequences encoding a xylonate dehydratase, and/or introducing one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and/or introducing one or more polynucleotide sequences encoding hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase;
• wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe lacking the exogenous introduced or endogenous overexpressed xylonolactonase, and any of the modifications above;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by:
• introducing one or more exogenous polynucleotide or sequences or overexpressing an endogenous polynucleotide or sequences encoding a xylonate dehydratase and introducing one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and/or introducing one or more polynucleotide sequences encoding hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase;
• wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe lacking the exogenous introduced or endogenous overexpressed xylonate dehydratase, any of the modifications above;
• or
• modifying a microbe coproducing MEG and one or more C3 compounds by:
• overexpressing one or more endogenous polynucleotide sequences encoding an acetyl-CoA synthetase;
• and disrupting one or more nucleic acid sequences encoding methylglyoxal synthase (mgsA), disrupting one or more nucleic acid sequences encoding glyoxylate carboligase (gcl) and/or;
• disrupting one or more endogenous polynucleotide sequences encoding an acetate kinase, and/or introducing one or more exogenous polynucleotide sequences encoding a xylonolactonase, and/or introducing one or more exogenous polynucleotide or sequences or overexpressing an endogenous polynucleotide or sequences encoding a xylonate dehydratase, and/or introducing one or more polynucleotide sequences encoding acetoacetyl CoA synthase, and/or introducing one or more polynucleotide sequences encoding hydroxymethylglutaryl-CoA synthase and hydroxymethylglutaryl-CoA lyase;
• disrupting one or more endogenous polynucleotide sequences encoding an ArcA regulator, and/or disrupting one or more endogenous polynucleotide sequences encoding a lysine acetyltransferase, and/or wherein the MEG and/or the one or more C3 compounds is produced at a faster titer, rate or exhibits an increased yield;
• as compared to a microbe lacking the endogenous overexpressed acetyl-CoA synthetase and any of the modifications above.