Methods for the economical production of biofuel precursor that is also a biofuel from biomass
AssigneeGevo, Inc.
InventorsAndrew C. Hawkins, David A. Glassner, Thomas Buelter, James Wade, Peter Meinhold, Matthew W. Peters, Patrick R. Gruber, William A. Evanko, Aristos A. Aristidou
Life of this patent
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Abstract
Methods for producing a biofuel precursor are provided. Also provided are biocatalysts that convert a feedstock to a biofuel precursor.
Claims240
A method of making a biofuel precursor, comprising:
- providing a biocatalyst selected to convert a feedstock into the biofuel precursor at a yield of at least 80 percent of theoretical, a productivity of at least 0.75 grams biofuel precursor per liter per hour, and a titer equivalent to a lower one of (i) a solubility limit of the biofuel precursor in water under the process conditions and (ii) 2% (w/w) of the biofuel precursor in water;
- providing the biocatalyst selected to have at least two properties from a. to l. as follows:
- a. the biocatalyst selected to convert at least two sugars, including each of (i) at least one of a six-carbon sugar and a six-carbon sugar oligomer, and (ii) at least one five-carbon sugar, derived from at least one of starch, cellulose, hemicellulose, and pectin into the biofuel precursor;
- b. the biocatalyst exhibiting a level of endotoxin toxicity or exotoxin toxicity, wherein the level of endotoxin or exotoxin toxicity in the biocatalyst has a median lethal dose (LD50) of at least 1000-fold more than the amount present in 1 kilogram of at least one of a DDG and a DDGS product;
- c. the biocatalyst containing no DNA markers;
- d. the biocatalyst operable to produce the biofuel precursor free of byproducts that would require additional processing steps for removal from the biofuel precursor;
- e. the biocatalyst operable at a pH value between about 2 to 10 to produce the biofuel precursor;
- f. the biocatalyst selected to have a recoverable productivity from a sudden change of about 1 pH unit from the first pH;
- g. the biocatalyst operable within a temperature range of about 20° C. to 60° C. to produce the biofuel precursor;
- h. the biocatalyst selected to have a recoverable productivity from a sudden change of about 5° C. from the first temperature;
- i. the biocatalyst operable in a medium comprising feedstock;
- j. the biocatalyst selected to have a growth rate of at least 0.2 per hour;
- k. providing a facultative anaerobic biocatalyst operable at dissolved oxygen concentrations across a range of about 0% to about 0.01% air saturation to produce the biofuel precursor;
- and
- l. providing an anaerobic biocatalyst selected to have a productivity that recovers from an exposure to more than 1% air saturation that lasts for up to three hours;
- and
- cultivating the biocatalyst in a culture medium until a recoverable quantity of the biofuel precursor is produced;
- and
- recovering the biofuel precursor.
A method of making a biofuel precursor, comprising:
- providing a biocatalyst selected to convert a feedstock into the biofuel precursor at a yield of at least 80 percent theoretical yield, a productivity of at least 0.75 grams biofuel precursor per liter per hour, and a titer equivalent to a lower one of (i) a solubility limit of the biofuel precursor in water under the process conditions and (ii) 2% (w/w) of the biofuel precursor in water;
- and
- cultivating the biocatalyst in a culture medium until a recoverable quantity of the biofuel precursor is produced;
- and
- recovering the biofuel precursor.
A method of making a biofuel precursor, the method comprising:
- a) providing a feedstock comprising a sugar selected from the group consisting of:
- i) a six-carbon sugar;
- ii) a six-carbon sugar oligomer;
- iii) a five-carbon sugar and
- iv) any combination of i) through iii),
- wherein the sugar is obtained from starch, cellulose, hemicellulose, or pectin;
- and
- b) contacting the feedstock of a) with a biocatalyst that converts the feedstock into a biofuel precursor at:
- i) a yield of at least 68 percent theoretical yield
- ii) a productivity of at least 0.75 grams biofuel precursor per liter per hour change claims;
- and
- iii) a lower one of (A) a solubility limit of the biofuel precursor in water under the process conditions and (B) 1.6% (w/v) of the biofuel precursor in water;
- and
- c) recovering the biofuel precursor.
A method of making a biofuel precursor, the method comprising:
- a) providing a feedstock comprising galactose obtained from starch, cellulose, hemicellulose, or pectin;
- and
- b) contacting the feedstock of a) with a biocatalyst that converts the feedstock into a biofuel precursor at:
- i) a yield of at least about 90 percent of the yield of the biocatalyst wherein the feedstock comprises glucose
- ii) a productivity of at least about 90 percent of the productivity of the biocatalyst wherein the feedstock comprises glucose;
- and
- iii) a titer of at least about 90 percent of the titer of the biocatalyst wherein the feedstock comprises glucose;
- and
- c) recovering the biofuel precursor.
A method of making a biofuel precursor, the method comprising:
- a) providing a feedstock comprising mannose obtained from starch, cellulose, hemicellulose, or pectin;
- and
- b) contacting the feedstock of a) with a biocatalyst that converts the feedstock into a biofuel precursor at:
- i) a yield of at least about 90 percent of the yield of the biocatalyst wherein the feedstock comprises glucose
- ii) a productivity of at least about 90 percent of the productivity of the biocatalyst wherein the feedstock comprises glucose;
- and
- iii a titer of at least about 90 percent of the titer of the biocatalyst wherein the feedstock comprises glucose;
- and
- c) recovering the biofuel precursor.
A method of making a biofuel precursor, the method comprising:
- a) providing a feedstock comprising xylose obtained from starch, cellulose, hemicellulose, or pectin;
- and
- b) contacting the feedstock of a) with a biocatalyst that converts the feedstock into a biofuel precursor at:
- i) a yield of at least about 69 percent of the yield of the biocatalyst wherein the feedstock comprises glucose
- ii) a productivity of at least about 56 percent of the productivity of the biocatalyst wherein the feedstock comprises glucose;
- and
- iii) a titer of at least about 59 percent of the titer of the biocatalyst wherein the feedstock comprises glucose;
- and
- c) recovering the biofuel precursor.
A method of making a biofuel precursor, the method comprising:
- a) providing a feedstock comprising arabinose obtained from starch, cellulose, hemicellulose, or pectin;
- and
- b) contacting the feedstock of a) with a biocatalyst that converts the feedstock into a biofuel precursor at:
- i) a yield of at least about 56 percent of the yield of the biocatalyst wherein the feedstock comprises glucose
- ii) a productivity of at least about 70 percent of the productivity of the biocatalyst wherein the feedstock comprises glucose;
- and
- iii) a titer of at least about 54 percent of the titer of the biocatalyst wherein the feedstock comprises glucose;
- and
- c) recovering the biofuel precursor.
A method of making a biofuel precursor, the method comprising:
- a) providing a feedstock comprising lactose;
- and
- b) contacting the feedstock of a) with a biocatalyst that converts the feedstock into a biofuel precursor at:
- i) a productivity of at least about 90 percent of the productivity of the biocatalyst wherein the feedstock comprises glucose;
- and
- ii) a titer of at least about 90 percent of the titer of the biocatalyst wherein the feedstock comprises glucose;
- and
- c) recovering the biofuel precursor.
A method of making a biofuel precursor, the method comprising:
- a) providing a feedstock comprising sucrose;
- and
- b) contacting the feedstock of a) with a biocatalyst that converts the feedstock into a biofuel precursor at:
- i) a productivity of at least about 50 percent of the productivity of the biocatalyst wherein the feedstock comprises glucose;
- and
- ii) a titer of at least about 50 percent of the titer of the biocatalyst wherein the feedstock comprises glucose;
- and
- c) recovering the biofuel precursor.
A method of making a biofuel precursor, the method comprising:
- a) providing a feedstock comprising a six carbon sugar obtained from starch, cellulose, hemicellulose, or pectin;
- and
- b) contacting the feedstock of a) with a biocatalyst that converts the feedstock into a biofuel precursor at:
- i) a yield of at least about 90 percent of the yield of the biocatalyst wherein the feedstock comprises glucose
- ii) a productivity of at least about 90 percent of the productivity of the biocatalyst wherein the feedstock comprises glucose;
- and
- iii) a titer of at least about 90 percent of the titer of the biocatalyst wherein the feedstock comprises glucose;
- and
- c) recovering the biofuel precursor.
A method of making a biofuel precursor, the method comprising:
- a) providing a feedstock comprising a five carbon sugar obtained from starch, cellulose, hemicellulose, or pectin;
- and
- b) contacting the feedstock of a) with a biocatalyst that converts the feedstock into a biofuel precursor at:
- i) a yield of at least about 55 percent of the yield of the biocatalyst wherein the feedstock comprises glucose
- ii) a productivity of at least about 55 percent of the productivity of the biocatalyst wherein the feedstock comprises glucose;
- and
- iii) a titer of at least about 55 percent of the titer of the biocatalyst wherein the feedstock comprises glucose;
- and
- c) recovering the biofuel precursor.
A method of producing a biofuel precursor comprising contacting a feedstock with a biocatalyst, wherein the method produces a biofuel precursor at a total titer of greater than about 22 g/L.
A method of producing a biofuel precursor comprising contacting a feedstock with a biocatalyst, wherein the method produces a biofuel precursor at a yield of greater than about 80 percent theoretical.
A method of producing a biofuel precursor comprising contacting a feedstock with a biocatalyst, wherein the method produces a biofuel precursor at a productivity of greater than about 2 g biofuel precursor per L per hour.
An isolated or recombinant biocatalyst comprising a recombinant biochemical pathway to produce isobutanol from fermentation of a suitable bio-mass, wherein the recombinant biochemical pathway comprises elevated activity of:
- a) a KARI as compared to a parental microorganism;
- and
- b) a ALS as compared to a parental microorganism,
- wherein the biocatalyst produces recoverable amounts of isobutanol.
An isolated or recombinant biocatalyst comprising a recombinant biochemical pathway to produce isobutanol from fermentation of a suitable bio-mass, wherein the recombinant biochemical pathway comprises decreased activity of:
- a) a DHAD as compared to a parental microorganism;
- and
- b) a kivd as compared to a parental microorganism,
- wherein the biocatalyst produces recoverable amounts of isobutanol.
An isolated or recombinant biocatalyst comprising a recombinant biochemical pathway to produce isobutanol from fermentation of a suitable bio-mass, wherein the recombinant biochemical pathway comprises decreased activity of:
- a) DHAD as compared to a parental microorganism;
- and increased activity of:
- b) ALS is broader as compared to a parental microorganism;
- and
- c) KARI as compared to a parental microorganism.
An isolated or recombinant biocatalyst comprising a genotype of E. coli BW25113, and further comprising a genotype of:
- ΔldhA-fnr::FRT, ΔadhE::FRT, Δfrd::FRT, Δpta::FRT, pflB::FRT, Δmdh::FRT, ΔaceF::FRT F′ (laclq+), pSA55, pSA69, wherein the isolated or recombinant biocatalyst produces isobutanol from a carbon source.
An isolated or recombinant biocatalyst comprising a genotype of E. coli B, and further comprising a genotype of:
- A adhE::FRT-kan-FRT, attB::(Sp+laclq+tetR+), pSA55, pGV1609), wherein the isolated or recombinant biocatalyst produces a metabolite comprising isobutanol from a carbon source.
An isolated or recombinant biocatalyst comprising a genotype of (E. coli BW25113, and further comprising a genotype of:
- ΔldhA-fnr::FRT, ΔadhE::FRT, Δfrd::FRT, Δpta::FRT, pflB::FRT, F′ (laclq+), pGV1655, pGV1698, wherein the isolated or recombinant biocatalyst produces isobutanol from a carbon source.
An isolated or recombinant biocatalyst comprising a genotype of E. coli BW25113, and further comprising a genotype of:
- ΔldhA-fnr::FRT, Δfrd::FRT, Δpta::FRT, F′ (laclq+), ΔadhE::[pLlacO1::kivd::ilvDco::FRT], ΔpflB::[pLlacO1::alsS::ilvCco::FRT] ΔsthA::[pLlacO1::pntA::pntB::FRT]), wherein the isolated or recombinant biocatalyst produces isobutanol from a carbon source.
An isolated or recombinant biocatalyst comprising a genotype of E. coli BW25113, and further comprising a genotype of:
- ΔldhA-fnr::FRT, ΔadhE::FRT, Δfrd::FRT, Δpta::FRT, pflB::FRT, F′ (laclq+), ΔilvC::[PLlacO1::kivd::ilvDco::FRT], pGV1698, wherein the isolated or recombinant biocatalyst produces isobutanol from a carbon source.
An isolated or recombinant biocatalyst comprising a genotype of E. coli BW25113, and further comprising a genotype of:
- ΔldhA-fnr::FRT, Δfrd::FRT, Δpta::FRT, pflB::FRT, F′ (laclq+), ΔadhE::[PLlacO1::kivd::ilvDco::FRT], pGV1698, wherein the isolated or recombinant biocatalyst produces isobutanol from a carbon source.
An isolated or recombinant biocatalyst comprising a genotype of E. coli BW25113, and further comprising a genotype of:
- ΔldhA-fnr::FRT, ΔadhE::FRT, Δfrd::FRT, ΔpflB::FRT, Δpta::FRT, F′ (laclq+), ΔilvC::[PLlacO1::kivd::ilvDco::FRT], ΔsthA::FRT, pGV1698, wherein the isolated or recombinant biocatalyst produces isobutanol from a carbon source.
An isolated or recombinant biocatalyst comprising a genotype of GEVO1748, and further comprising a genotype of:
- pGV1745, pGV1698, wherein the isolated or recombinant biocatalyst produces isobutanol from a carbon source.
An isolated or recombinant biocatalyst comprising a genotype of E. coli BW25113, and further comprising a genotype of:
- ΔldhA-fnr::FRT, Δfrd::FRT, Δpta::FRT, F′ (laclq+), ΔadhE::[pLlacO1::kivd::ilvDco::FRT], pflB::[pLlacO1::alsS::ilvCco::FRT], wherein the isolated or recombinant biocatalyst produces isobutanol from a carbon source.
An isolated or recombinant biocatalyst comprising a genotype of E. coli BW25113, and further comprising a genotype of:
- ΔldhA-fnr::FRT, Δfrd::FRT, Δpta::FRT, ΔadhE::[pLlacO1::kivd::ilvDco::FRT], ΔpflB::[pLlacO1::alsS::ilvCco::FRT] ΔsthA::[pLlacO1::pntA::pntB::FRT], wherein the isolated or recombinant biocatalyst produces isobutanol from a carbon source.
An isolated or recombinant nucleic acid comprising pLlacO1::alsS::ilvC::ilvD, p15A, and Cm.
An isolated or recombinant nucleic acid comprising pLlacO1::
- kivd::ilvDco, pSC101, and Kan.
An isolated or recombinant nucleic acid comprising PLlacO1::alsS::ilvCco, ColE1, and Amp.
An isolated or recombinant nucleic acid comprising pLlacO1::pntAB, pSC101, and Kan.
A biocatalyst selected to convert a feedstock into the biofuel precursor at a yield of at least 80 percent theoretical yield, a productivity of at least 0.75 grams biofuel precursor per liter per hour, and a titer equivalent to a lower one of (i) a solubility limit of the biofuel precursor in water under the process conditions and (ii) 2% (w/w) of the biofuel precursor in water;
- providing the biocatalyst selected to have the following properties:
- a. the biocatalyst operable to produce the biofuel precursor free of byproducts that would require additional processing steps for removal from the biofuel precursor;
- b. the biocatalyst selected to have a growth rate of at least 0.2 per hour.
Dependencies are inferred from the claim text.