Methods of producing four carbon molecules

Abstract

Disclosed are methods for producing butadiene from one or more of several diverse feedstocks including bioderived feedstocks, renewable feedstocks, petrochemical feedstocks and natural gas.

1 . A method for biosynthesizing butadiene comprising:
 fermenting in an organism in a fermentation broth, a C4-precursor, wherein said C4-precursor comprises butanol, butanediol, or both, or derivatives thereof;   converting C4-precursor in the fermentation broth into butenol by:
 (a) converting butanediol to butenol, or 
 (b) converting butanol to butenol; 
   converting the butenol into 1,3 butadiene; and   optionally isolating said 1,3-butadiene.   
     
     
         2 . The method of  claim 1 , wherein the fermentation broth comprises a biologically derived fermentable feedstock or a nonbiologically derived fermentable feedstock. 
     
     
         3 . The method of  claim 2 , wherein the biologically derived feedstock comprises glycerol, synthesis gas, synthesis gas from biomass, sugar, sugar from a food stuff, sugar from a non-food stuff, or combinations thereof. 
     
     
         4 . The method of  claim 3 , wherein the sugar is sucrose, glucose, or combinations thereof. 
     
     
         5 . The method of  claim 3 , wherein the sugar from a non-food stuff is cellulosic or hemi-cellulosic sugar or combinations thereof. 
     
     
         6 . (canceled) 
     
     
         7 . The method of  claim 2 , wherein the non-biologically derived feedstock is synthesis gas from coal, natural gas, combustion off-gases, municipal waste, petrochemical, or combinations thereof. 
     
     
         8 . (canceled) 
     
     
         9 . The method of  claim 1 , wherein the organism is a prokaryote or eukaryote. 
     
     
         10 . The method of  claim 1 , wherein the C4 precursor is a hydroxylated C4 molecule. 
     
     
         11 . The method of  claim 1 , wherein converting the butanediol to butenol requires at least one polypeptide having an activity of at least one enzyme. 
     
     
         12 . The method of  claim 11 , wherein the at least one enzyme is a dehydratase, desaturase or hydrolase enzyme. 
     
     
         13 . The method of  claim 11 , wherein the at least one enzyme is a polypeptide having an activity of an enzyme EC 4.2.1, EC 1.14.19, EC 1.14.99, EC 1.3.1.35, cytochrome P450, or EC 1.14.11.22. 
     
     
         14 - 17 . (canceled) 
     
     
         18 . The method of  claim 1 , wherein the butanediol is 1,4-butanediol, 1,3-butanediol, or 2,3-butanediol or derivatives thereof. 
     
     
         19 . The method of  claim 1 , wherein the butenol is 1-buten-1-ol, 1-buten-2-ol, 1-buten-3-ol, 1-buten-4-ol, 2-buten-1-ol, 2-buten-2-ol, 2-buten-3-ol, or 2-buten-4-ol or derivatives thereof. 
     
     
         20 . (canceled) 
     
     
         21 . The method of  claim 1 , wherein the organism expresses at least one polypeptide having an enzyme activity capable of converting the C4 precursors to butadiene. 
     
     
         22 . The method of  claim 21 , wherein the organism is recombinantly engineered to express the polypeptide having the enzyme activity. 
     
     
         23 . The method of  claim 21 , wherein the polypeptide has the enzyme activity of a dehydratase, a desaturase, or both. 
     
     
         24 - 35 . (canceled) 
     
     
         36 . A method for biosynthesising butadiene from a petrochemical feedstock comprising:
 obtaining a petrochemical feedstock comprising butane;   fermenting butane in the presence of an organism to produce a fermentation broth comprising butene; and   fermenting the butene in the presence of an organism to produce 1,3-butadiene in the fermentation broth; and   optionally obtaining the 1,3-butadiene.   
     
     
         37 . (canceled) 
     
     
         38 . A microorganism genetically modified to produce 1,3-butadiene. 
     
     
         39 . The microorganism of  claim 38  wherein the organism is a prokaryote or eukaryote. 
     
     
         40 . The microorganism of  claim 38  recombinantly engineered to express at least one polypeptide having an activity of an enzyme capable of converting C4 precursors to butadiene. 
     
     
         41 . The microorganism of  claim 40  wherein the polypeptide having the activity of the enzyme is a dehydratase, desaturase or hydrolase enzyme. 
     
     
         42 . The microorganism of  claim 41  which comprises a heterologous nucleic acid sequence encoding a polypeptide having a enzyme activity of an enzyme EC 4.2.1, EC 1.14.19, EC 1.14.99, EC 1.3.1.35, cytochrome P450, or EC 1.14.11.22. 
     
     
         43 . The microorganism of  claim 41  which comprises heterologous nucleic acid sequences encoding a polypeptide having an enzyme activity of a dehydratase enzyme classified under EC 4.2.1.- and a desaturase enzyme selected from the group consisting of cytochrome P450 enzymes, clavaminate synthase 2 enzymes and enzymes classified under EC 1.14.19-, EC 1.14.99-, EC 1.3.1-. 
     
     
         44 . A bio-derived, bio-based or fermentation-derived product, wherein said product comprises:
 i. a composition comprising at least one bio-derived, bio-based or fermentation-derived compound obtained according to  claim 1  or any combination thereof,   ii. a bio-derived, bio-based or fermentation-derived polymer comprising the bio-derived, bio-based or fermentation-derived composition or compound of i., or any combination thereof,   iii. a bio-derived, bio-based or fermentation-derived resin comprising the bio-derived, bio-based or fermentation-derived compound or bio-derived, bio-based or fermentation-derived composition of i. or any combination thereof or the bio-derived, bio-based or fermentation-derived polymer of ii. or any combination thereof,   iv. a molded substance obtained by molding the bio-derived, bio-based or fermentation-derived polymer of ii. or the bio-derived, bio-based or fermentation-derived resin of iii., or any combination thereof,   v. a bio-derived, bio-based or fermentation-derived formulation comprising the bio-derived, bio-based or fermentation-derived composition of i., bio-derived, bio-based or fermentation-derived compound of i., bio-derived, bio-based or fermentation-derived polymer of ii., bio-derived, bio-based or fermentation-derived resin of iii., or bio-derived, bio-based or fermentation-derived molded substance of iv, or any combination thereof, or   vi. a bio-derived, bio-based or fermentation-derived semi-solid or a non-semi-solid stream, comprising the bio-derived, bio-based or fermentation-derived composition of i., bio-derived, bio-based or fermentation-derived compound of i., bio-derived, bio-based or fermentation-derived polymer of ii., bio-derived, bio-based or fermentation-derived resin of iii., bio-derived, bio-based or fermentation-derived formulation of v., or bio-derived, bio-based or fermentation-derived molded substance of iv., or any combination thereof.   
     
     
         45 . A non-naturally occurring biochemical network comprising at least one substrate of  FIG. 1 ,  FIG. 2  or  FIG. 3  and at least one exogenous nucleic acid encoding a polypeptide having the activity of at least one enzyme of  FIG. 1 ,  FIG. 2  or  FIG. 3 . 
     
     
         46 . A non-naturally occurring composition, comprising at least one substrate of  FIG. 1 ,  FIG. 2  or  FIG. 3 , wherein said substrate is optionally bio-based, bio-derived or fermentation derived; at least one exogenous nucleic acid encoding a polypeptide having the activity of at least one enzyme of  FIG. 1 ,  FIG. 2  or  FIG. 3 ; and at least one bio-based, bio-derived or fermentation derived product of  FIG. 1 ,  FIG. 2  or  FIG. 3 .