Programmed microorganisms to attenuate a disease

A recombinant microbe producing etoposide, or its derivatives, comprising genes encoding phenyl alanine ammonia-lyase (PAL), cinnamate-4-hydroxylate (C4H), 4-coumaroyl CoA-ligase (4CL), hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase (HCT), p-coumaroyl quinate 3′-hydroxylase (HCTC3H), caffeoyl CoA O-methyltransferase (CCoAOMT), bifunctional pineresionl-lariciresinol reductase (DIRPLR), secoisolariciresinol dehydrogenase (SDH), cytochrome P450 oxidoreductase CYP719, O-methyltransferase (OMT), cytochrome P450 oxidoreductase CYP71, 2-oxoglutarate/Fe(II)-dependent dioxygenase (2-ODD), cytochrome P450 oxidoreductase CYP82D, UDP glucosyl transferase, and 2-Deoxy-d-ribose-5-phosphate aldolase.

A recombinant microbe comprising genes encoding phenyl alanine ammonia-lyase (PAL), cinnamate-4-hydroxylate 4-coumaroyl CoA-ligase fusion (C4H4CL), hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase p-coumaroyl quinate 3′-hydroxylase fusion (HCTC3H), caffeoyl CoA O-methyltransferase (CCoAOMT), bifunctional pineresionl-lariciresinol reductase (DIRPLR), secoisolariciresinol dehydrogenase (SDH), O-methyltransferase (OMT), 2-oxoglutarate/Fe(II)-dependent dioxygenase (2-ODD), 2-Deoxy-d-ribose-5-phosphate aldolase, Cytochrome P450 oxidoreductase CYP719, Cytochrome P450 oxidoreductase CYP71, Cytochrome P450 oxidoreductase CYP82D, and UDP glucosyl transferase, and a protein transporter, wherein the recombinant microbe secretes etoposide, or its derivatives.

A recombinant microbe comprising genes encoding phenyl alanine ammonia-lyase (PAL), cinnamate-4-hydroxylate 4-coumaroyl CoA-ligase fusion (C4H4CL), hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase p-coumaroyl quinate 3′-hydroxylase fusion (HCTC3H), caffeoyl CoA O-methyltransferase (CCoAOMT), bifunctional pineresionl-lariciresinol reductase (DIRPLR), secoisolariciresinol dehydrogenase (SDH), O-methyltransferase (OMT), 2-oxoglutarate/Fe(II)-dependent dioxygenase (2-ODD), 2-Deoxy-d-ribose-5-phosphate aldolase, Cytochrome P450 oxidoreductase CYP719, Cytochrome P450 oxidoreductase CYP71, Cytochrome P450 oxidoreductase CYP82D, and UDP glucosyl transferase, and a protein transporter, wherein the recombinant microbe secretes etoposide, or its derivatives, and wherein the expression of the genes is under the control of at least one regulatory circuit.

A recombinant microbe comprising genes encoding phenyl alanine ammonia-lyase (PAL), cinnamate-4-hydroxylate 4-coumaroyl CoA-ligase fusion (C4H4CL), hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase p-coumaroyl quinate 3′-hydroxylase fusion (HCTC3H), caffeoyl CoA O-methyltransferase (CCoAOMT), bifunctional pineresionl-lariciresinol reductase (DIRPLR), secoisolariciresinol dehydrogenase (SDH), O-methyltransferase (OMT), 2-oxoglutarate/Fe(II)-dependent dioxygenase (2-ODD), 2-Deoxy-d-ribose-5-phosphate aldolase, Cytochrome P450 oxidoreductase CYP719, Cytochrome P450 oxidoreductase CYP71, Cytochrome P450 oxidoreductase CYP82D, and UDP glucosyl transferase, and a protein transporter, wherein the recombinant microbe secretes etoposide, or its derivatives, and wherein the expression of the genes is under the control of a hypoxia-responsive regulatory circuit.

A recombinant microbe comprising genes encoding phenyl alanine ammonia-lyase (PAL), cinnamate-4-hydroxylate 4-coumaroyl CoA-ligase fusion (C4H4CL), hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase p-coumaroyl quinate 3′-hydroxylase fusion (HCTC3H), caffeoyl CoA O-methyltransferase (CCoAOMT), bifunctional pineresionl-lariciresinol reductase (DIRPLR), secoisolariciresinol dehydrogenase (SDH), O-methyltransferase (OMT), 2-oxoglutarate/Fe(II)-dependent dioxygenase (2-ODD), 2-Deoxy-d-ribose-5-phosphate aldolase, Cytochrome P450 oxidoreductase CYP719, Cytochrome P450 oxidoreductase CYP71, Cytochrome P450 oxidoreductase CYP82D, and UDP glucosyl transferase, and a protein transporter, wherein the recombinant microbe secretes etoposide, or its derivatives, and wherein the expression of the genes is under the control of a nitric oxide-responsive regulatory circuit.

A recombinant microbe comprising genes encoding phenyl alanine ammonia-lyase (PAL), cinnamate-4-hydroxylate 4-coumaroyl CoA-ligase fusion (C4H4CL), hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase p-coumaroyl quinate 3′-hydroxylase fusion (HCTC3H), caffeoyl CoA O-methyltransferase (CCoAOMT), bifunctional pineresionl-lariciresinol reductase (DIRPLR), secoisolariciresinol dehydrogenase (SDH), O-methyltransferase (OMT), 2-oxoglutarate/Fe(II)-dependent dioxygenase (2-ODD), 2-Deoxy-d-ribose-5-phosphate aldolase, Cytochrome P450 oxidoreductase CYP719, Cytochrome P450 oxidoreductase CYP71, Cytochrome P450 oxidoreductase CYP82D, and UDP glucosyl transferase, and a protein transporter, wherein the recombinant microbe secretes etoposide, or its derivatives, and wherein the expression of the genes is under the control of an arabinose-responsive regulatory circuit.

A recombinant vector comprising at least one nucleic acid fragment encoding phenyl alanine ammonia-lyase (PAL), Cinnamate-4-hydroxylate (C4H), 4-coumaroyl CoA-ligase (4CL), hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase (HCT), p-coumaroyl quinate 3′-hydroxylase (C3H), caffeoyl CoA O-methyltransferase (CCoAOMT), bifunctional pineresionl-lariciresinol reductase (DIRPLR), secoisolariciresinol dehydrogenase (SDH), O-methyltransferase (OMT), 2-oxoglutarate/Fe(II)-dependent dioxygenase (2-ODD), 2-Deoxy-d-ribose-5-phosphate aldolase, Cytochrome P450 oxidoreductase CYP719, Cytochrome P450 oxidoreductase CYP71, Cytochrome P450 oxidoreductase CYP82D, and UDP glucosyl transferase, at least one gene encoding a protein transporter selected from the group consisting of ATP-Binding Cassette (ABC) transporter, Major Facilitator Superfamily (MFS) transporters, SMR (small multidrug resistant) family, RND (Resistance-Nodulation-Cell Division) family, and the MATE (multidrug and toxic compound extrusion) family, and at least one regulatory circuit selected from the group consisting of nitric oxide (NO) operon, arabinose (AraC) operon, fumarate and nitrate reductase (FNR) operon, thiosulphate-responsive regulatory circuit, and tetrathionate-responsive regulatory circuit.