Organisms for the production of 1,3-butanediol

Abstract

A non-naturally occurring microbial organism includes a microbial organism having a 1,3-butanediol (1,3-BDO) pathway having at least one exogenous nucleic acid encoding a 1,3-BDO pathway enzyme expressed in a sufficient amount to produce 1,3-BDO. The pathway includes an enzyme selected from a 2-amino-4-ketopentanoate (AKP) thiolase, an AKP dehydrogenase, a 2-amino-4-hydroxypentanoate aminotransferase, a 2-amino-4-hydroxypentanoate oxidoreductase (deaminating), a 2-oxo-4-hydroxypentanoate decarboxylase, a 3-hydroxybutyraldehyde reductase, an AKP aminotransferase, an AKP oxidoreductase (deaminating), a 2,4-dioxopentanoate decarboxylase, a 3-oxobutyraldehyde reductase (ketone reducing), a 3-oxobutyraldehyde reductase (aldehyde reducing), a 4-hydroxy-2-butanone reductase, an AKP decarboxylase, a 4-aminobutan-2-one aminotransferase, a 4-aminobutan-2-one oxidoreductase (deaminating), a 4-aminobutan-2-one ammonia-lyase, a butenone hydratase, an AKP ammonia-lyase, an acetylacrylate decarboxylase, an acetoacetyl-CoA reductase (CoA-dependent, aldehyde forming), an acetoacetyl-CoA reductase (CoA-dependent, alcohol forming), an acetoacetyl-CoA reductase (ketone reducing), a 3-hydroxybutyryl-CoA reductase (aldehyde forming), a 3-hydroxybutyryl-CoA reductase (alcohol forming), a 4-hydroxybutyryl-CoA dehydratase, and a crotonase. A method for producing 1,3-BDO, includes culturing such microbial organisms under conditions and for a sufficient period of time to produce 1,3-BDO.

1 - 56 . (canceled) 
     
     
         57 . A non-naturally occurring microbial organism having a 1,3-butanediol (1,3-BDO) pathway, wherein said microbial organism comprises at least one exogenous nucleic acid encoding a 3-hydroxybutyryl-CoA reductase (aldehyde forming) expressed in a sufficient amount to produce 1,3-BDO. 
     
     
         58 . The non-naturally occurring microbial organism of  claim 57 , wherein said 3-hydroxybutyryl-CoA reductase (aldehyde forming) is encoded by one or more genes selected from the group consisting of acr1, sucD, bphG, bld, adhE, Msed_0709, mcr, asd-2, Saci_2370, Ald, and eutE. 
     
     
         59 . The non-naturally occurring microbial organism of  claim 57 , wherein said microbial organism comprises two exogenous nucleic acids, each encoding a 1,3-BDO pathway enzyme. 
     
     
         60 . The non-naturally occurring microbial organism of  claim 57 , wherein said microbial organism comprises three exogenous nucleic acids, each encoding a 1,3-BDO pathway enzyme. 
     
     
         61 . The non-naturally occurring microbial organism of  claim 57 , wherein said microbial organism comprises four exogenous nucleic acids, each encoding a 1,3-BDO pathway enzyme. 
     
     
         62 . The non-naturally occurring microbial organism of  claim 57 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid. 
     
     
         63 . The non-naturally occurring microbial organism of  claim 57 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium. 
     
     
         64 . The non-naturally occurring microbial organism of  claim 57 , wherein said 1,3-BDO pathway comprises a set of 1,3-BDO pathway enzymes, said set of 1,3-BDO pathway enzymes comprises (1) an acetoacetyl-CoA reductase (ketone reducing); (2) a 3-hydroxybutyryl-CoA reductase (aldehyde forming); and (3) a 3-hydroxybutyraldehyde reductase. 
     
     
         65 . The non-naturally occurring microbial organism of  claim 57 , wherein said 1,3-BDO pathway comprises a set of 1,3-BDO pathway enzymes, said set of 1,3-BDO pathway enzymes comprises (1) a 4-hydroxybutyryl-CoA dehydratase; (2) a crotonase; (3) a 3-hydroxybutyryl-CoA reductase (aldehyde forming); and (4) a 3-hydroxybutyraldehyde reductase. 
     
     
         66 . The non-naturally occurring microbial organism of  claim 64 , wherein said acetoacetyl-CoA reductase (ketone reducing) is encoded by one or more genes selected from the group consisting of thrA, akthr2, hom6, hom1, hom2, fadB, fadJ, Hbd2, Hbd1, hbd, HSD17B10, phbB, phaB, Msed_1423, Msed_0399, Msed_0389, Msed_1993, adh, adhA, adh-A, mdh, ldhA, ldh, and bdh. 
     
     
         67 . The non-naturally occurring microbial organism of  claim 65 , wherein said 4-hydroxybutyryl-CoA dehydratase is encoded by one or more genes selected from the group consisting of fumA, fumB, fumC, fumH, fum1, MmcB, MmcC, hmd, BACCAP_02294, ANACOL_02527, NtherDRAFT_2368, dmdA, dmdB, crt, crt1, ech paaA, paaB, phaA, phaB, maoC, paaF, paaG, abfD, Msed_1220, fadA, fadB, fadI, fadJ, and fadR. 
     
     
         68 . The non-naturally occurring microbial organism of  claim 65 , wherein said crotonase is encoded by one or more genes selected from the group consisting of fumA, fumB, fumC, fumH, fum1, MmcB, MmcC, hmd, BACCAP_02294, ANACOL_02527, NtherDRAFT_2368, dmdA, dmdB, crt, crt1, ech paaA, paaB, phaA, phaB, maoC, paaF, paaG, abfD, Msed_1220, fadA, fadB, fadI, fadJ, and fadR. 
     
     
         69 . The non-naturally occurring microbial organism of  claim 57 , wherein said microbial organism is a bacteria. 
     
     
         70 . The non-naturally occurring microbial organism of  claim 69 , wherein said bacteria is  E. coli.    
     
     
         71 . A culture medium comprising the non-naturally occurring microbial organism of  claim 57 . 
     
     
         72 . The culture medium of  claim 71  further comprising 1,3-BDO. 
     
     
         73 . A method for producing 1,3-BDO, comprising culturing the non-naturally occurring microbial of  claim 57  under conditions and for a sufficient period of time to produce 1,3-BDO. 
     
     
         74 . The method of  claim 73  further comprising separating 1,3-BDO from other components in the culture. 
     
     
         75 . The method of  claim 74 , wherein the separating comprises extraction, continuous liquid-liquid extraction, pervaporation, membrane filtration, membrane separation, reverse osmosis, electrodialysis, distillation, crystallization, centrifugation, extractive filtration, ion exchange chromatography, size exclusion chromatography, absorption chromatography, or ultrafiltration. 
     
     
         76 . The method of  claim 75 , wherein the separating comprises distillation. 
     
     
         77 . The method of  claim 74 , further comprising converting the 1,3-BDO to another compound.