Microorganisms and methods for producing (3R)-hydroxybutyl (3R)-hydroxybutyrate

Patent metadata

Field	Value
Publication number	US-11814664-B2
Application number	US-201414893510-A
Country	US
Kind code	B2
Filing date	May 23, 2014
Priority date	May 24, 2013
Publication date	Nov 14, 2023
Grant date	Nov 14, 2023

Abstract

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.

First claim

What is claimed is:

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.

Assignees

• Genomatica Inc

Inventors

• Adelstein Benjamin
• Osterhout Robin E
• Burk Mark J

Classifications

• C12P7/62 Primary

  Carboxylic acid esters · CPC title

• A23L33/135

  Bacteria or derivatives thereof, e.g. probiotics · CPC title

• A61K31/22

  of acyclic acids, e.g. pravastatin · CPC title

• A61K35/66

  Microorganisms or materials therefrom (fungi, yeasts or candida A61K36/06) · CPC title

• A61K36/064

  Saccharomycetales, e.g. baker's yeast · CPC title