Microorganisms and methods for producing (3R)-hydroxybutyl (3R)-hydroxybutyrate
Provided herein are non-naturally occurring microbial organisms having a pathway for production of (3R)-hydroxybutyl (3R)-hydroxybutyrate, wherein the organism can further include a (R)-1,3-butanediol pathway, a (3R)-hydroxybutyrate pathway, a (3R)-hydroxybutyryl-CoA pathway, an acetoacetate pathway, an acetoacetyl-CoA pathway, a (3R)-hydroxybutyl-ACP pathway, or an acetoacetyl-ACP pathway. Additionally provided are methods and processes for producing and isolating (3R)-hydroxybutyl (3R)-hydroxybutyrate using the microbial organisms, and various compositions having the (3R)-hydroxybutyl (3R)-hydroxybutyrate. Still further provided are methods of treating or preventing a disease, disorder or condition using the (3R)-hydroxybutyl (3R)-hydroxybutyrate produced by the microbial organisms of the invention.
1. A non-naturally occurring microbial organism, said microbial organism having a (3R)-hydroxybutyl (3R)-hydroxybutyrate pathway,
wherein said (3R)-hydroxybutyl (3R)-hydroxybutyrate pathway comprises one or more (3R)-hydroxybutyl (3R)-hydroxybutyrate pathway enzymes selected from:
(1) 1A;
(2) 1B;
(3) 1C and 1E;
(4) 1D and 1E;
(5) 1F; and
(6) 1G and 1E,
wherein 1A is a (3R)-hydroxybutyl (3R)-hydroxybutyrate ester forming enzyme, wherein 1B is a (3R)-hydroxybutyryl-CoA:(R)-1,3-butanediol alcohol transferase, wherein 1C is a (3R)-hydroxybutyl 3-oxobutyrate ester forming enzyme, wherein 1D is an acetoacetyl-CoA:(R)-1,3-butanediol alcohol transferase, wherein 1E is a (3R)-hydroxybutyl 3-oxobutyrate reductase, wherein 1F is a (3R)-hydroxybutyryl-ACP:(R)-1,3-butanediol ester synthase, wherein 1G is an acetoacetyl-ACP:(R)-1,3-butanediol ester synthase,
wherein the (3R)-hydroxybutyl (3R)-hydroxybutyrate pathway enzyme is encoded by at least one exogenous nucleic acid and is expressed in a sufficient amount to produce (3R)-hydroxybutyl (3R)-hydroxybutyrate.
2. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the enzymes of at least one of the pathways selected from (1)-(6).
3. The non-naturally occurring microbial organism of claim 1 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.
4. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism is a species of bacteria, yeast, or fungus.
5. A method for producing (3R)-hydroxybutyl (3R)-hydroxybutyrate, comprising culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce (3R)-hydroxybutyl (3R)-hydroxybutyrate.
6. A non-naturally occurring microbial organism, said microbial organism having a (3R)-hydroxybutyl (3R)-hydroxybutyrate pathway,
wherein said (3R)-hydroxybutyl (3R)-hydroxybutyrate pathway comprises one or more (3R)-hydroxybutyl (3R)-hydroxybutyrate pathway enzymes selected from:
(1) 1A;
(2) 1B;
(3) 1C and 1E;
(4) 1D and 1E;
(5) 1F; and
(6) 1G and 1E,
wherein 1A is a (3R)-hydroxybutyl (3R)-hydroxybutyrate ester forming enzyme, wherein 1B is a (3R)-hydroxybutyryl-CoA:(R)-1,3-butanediol alcohol transferase, wherein 1C is a (3R)-hydroxybutyl 3-oxobutyrate ester forming enzyme, wherein 1D is an acetoacetyl-CoA:(R)-1,3-butanediol alcohol transferase, wherein 1E is a (3R)-hydroxybutyl 3-oxobutyrate reductase, wherein 1F is a (3R)-hydroxybutyryl-ACP:(R)-1,3-butanediol ester synthase, wherein 1G is an acetoacetyl-ACP:(R)-1,3-butanediol ester synthase,
wherein the (3R)-hydroxybutyl (3R)-hydroxybutyrate pathway enzyme is encoded by at least one exogenous nucleic acid.