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US 2017/0121748 A1Published application
Microbial polycultures and methods of use thereof
AssigneeRensselaer Polytechnic Institute
InventorsMattheos KOFFAS, John Andrew JONES
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Abstract
Disclosed herein are novel microbial polycultures of two or more cell strains, capable of producing flavanones, flavonoids, and anthocyanidin-3-O-glucosides, and methods of use thereof. Also disclosed is a microbial cell capable of producing phenylpropanoic acids, and methods of use thereof.
Claims38
A method of producing a product compound in a microbial polyculture;
- wherein, optionally, the microbial polyculture comprises a TAL module cell comprising an exogenous gene encoding for a tyrosine ammonia lyase (TAL);
- wherein, optionally, the microbial polyculture comprises a C5 module cell comprising an exogenous gene encoding for a 4-coumaroyl-CoA ligase (4CL), an exogenous gene encoding for a chalcone synthase (CHS), an exogenous gene encoding for a chalcone isomerase (CHI), and wherein, optionally, the C5 module cell further comprises an exogenous gene encoding for malonyl-CoA synthetase (MatB) and an exogenous gene encoding for putative dicarboxylate carrier protein (MatC);
- wherein, optionally, the microbial polyculture further comprises a p168 module cell comprising an exogenous gene encoding for a flavanone 3β-hydroxylase (F3H), an exogenous gene encoding for a dihydroflavonol 4-reductase (DFR), and an exogenous gene encoding for a leucoanthocyanidin reductase (LAR);
- and
- wherein, optionally, the microbial polyculture further comprises an Antho module cell comprising an exogenous gene encoding for an anthocyanidin synthase (ANS) and an exogenous gene encoding for a 3-glucosyl transferase (3GT);
- the method comprising:
- providing a substrate to the microbial polyculture;
- culturing the microbial polyculture under conditions permitting synthesis of the product compound by the microbial polyculture;
- and
- isolating the product compound synthesized by the microbial polyculture;
- with a proviso that:
- the microbial polyculture comprises the TAL module cell and the C5 module cell, the substrate is glucose, glycerol, or a combination thereof, and the product compound is a flavanone;
- or
- the microbial polyculture comprises the C5 module cell and the p168 module cell, the substrate is a phenylpropanoic acid, and the product compound is a flavonoid;
- wherein, when the C5 module cell comprises an exogenous gene encoding for malonyl-CoA synthetase (MatB) and an exogenous gene encoding for putative dicarboxylate carrier protein (MatC), the substrate is a phenylpropanoic acid, malonate, or a combination thereof and the product is a flavonoid;
- or
- the microbial polyculture comprises the p168 module cell and the Antho module cell, the substrate is a flavanone, and the product compound is an anthocyanidin-3-O-glucoside;
- or
- the microbial polyculture comprises the TAL module cell, the C5 module cell, and the p168 module cell, the substrate is glucose, glycerol, or a combination thereof, and the product compound is a flavonoid;
- or
- the microbial polyculture comprises the C5 module cell, the p168 module cell, and the Antho module cell, the substrate is a phenylpropanoic acid, and the product compound is an anthocyanidin-3-O-glucoside;
- wherein, when the C5 module cell comprises an exogenous gene encoding for malonyl-CoA synthetase (MatB) and an exogenous gene encoding for putative dicarboxylate carrier protein (MatC), the substrate is phenylpropanoic acid, malonate, or a combination thereof and the product is an anthocyanidin-3-O-glucoside;
- or
- the microbial polyculture comprises the TAL module cell, the C5 module cell, the p168 module cell, and the Antho module cell, the substrate is glucose, glycerol, or a combination thereof, and the product compound is an anthocyanidin-3-O-glucoside.
21 more elements
A microbial polyculture, comprising:
- optionally, a TAL module cell comprising an exogenous gene encoding for a tyrosine ammonia lyase (TAL);
- optionally, a C5 module cell comprising an exogenous gene encoding for a 4-coumaroyl-CoA ligase (4CL), an exogenous gene encoding for a chalcone synthase (CHS), an exogenous gene encoding for a chalcone isomerase (CHI), and wherein, optionally, the C5 module cell further comprises an exogenous gene encoding for malonyl-CoA synthetase (MatB) and an exogenous gene encoding for putative dicarboxylate carrier protein (MatC);
- optionally, a p168 module cell comprising an exogenous gene encoding for a flavanone 3β-hydroxylase (F3H), an exogenous gene encoding for a dihydroflavonol 4-reductase (DFR), and an exogenous gene encoding for a leucoanthocyanidin reductase (LAR);
- and
- optionally, an Antho module cell comprising an exogenous gene encoding for an anthocyanidin synthase (ANS) and an exogenous gene encoding for a 3-glucosyl transferase (3GT);
- with a proviso that:
- the microbial polyculture comprises the TAL module cell and the C5 module cell;
- or
- the microbial polyculture comprises the C5 module cell and the p168 module cell;
- or
- the microbial polyculture comprises the p168 module cell and the Antho module cell;
- or
- the microbial polyculture comprises the TAL module cell, the C5 module cell, and the p168 module cell;
- or
- the microbial polyculture comprises the C5 module cell, the p168 module cell, and the Antho module cell;
- or
- the microbial polyculture comprises the TAL module cell, the C5 module cell, the p168 module cell, and the Antho module cell.
14 more elements
A method of producing a phenylpropanoic acid in a TAL module cell, wherein the TAL module cell is a microbial cell comprising an exogenous gene encoding for a tyrosine ammonia lyase (TAL);
- the method comprising:
- providing a substrate to the TAL module cell, wherein the substrate comprises glucose, glycerol, or a combination thereof;
- culturing the TAL module cell under conditions permitting synthesis of the phenylpropanoic acid by the TAL module cell;
- and
- isolating the phenylpropanoic acid synthesized by the TAL module cell.
2 more elements
A TAL module cell, wherein the TAL module cell is a microbial cell comprising an exogenous gene encoding for a tyrosine ammonia lyase (TAL).
Dependencies are inferred from the claim text.