Production of beta-glucosidase, hemicellulase and ligninase in E1 and FLC-cellulase-transgenic plants
First Claim
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1. A transgenic plant capable of expressing one or more cell wall degrading enzymes comprising:
- (a) at least one DNA comprising a cell wall degrading enzyme coding region operably linked to a nucleotide sequence encoding a signal peptide directing the cell wall degrading enzyme encoded by the DNA to a plastid or apoplast of the transgenic plant; and
(b) at least one DNA comprising a flowering locus c gene coding region operably linked to a constitutive promoter, wherein the transgenic plant expresses the one or more cell wall degrading enzymes and a transcription factor encoded by the flowering locus c gene that delays flowering while increasing biomass and enabling isolation of increased amounts of the hydrolyzing enzyme from the transgenic plant as compared to a non-transgenic plant from which the transgenic plant is derived.
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Abstract
The present invention provides transgenic plants expressing one or more cell wall degrading enzymes that can degrade lignocellulose to fermentable sugars. These fermentable sugars can further be fermented to ethanol or other products. The enzymes are directed to the plastids or the apoplasts or the transgenic plant for storage. When the transgenic plants are harvested, the plants are ground to release the enzymes which then are used to degrade the lignocellulose of plant material to produce the fermentable sugars. The transgenic plants express the flowering locus c gene so that flowering is delayed and the plant biomass is increased.
68 Citations
35 Claims
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1. A transgenic plant capable of expressing one or more cell wall degrading enzymes comprising:
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(a) at least one DNA comprising a cell wall degrading enzyme coding region operably linked to a nucleotide sequence encoding a signal peptide directing the cell wall degrading enzyme encoded by the DNA to a plastid or apoplast of the transgenic plant; and
(b) at least one DNA comprising a flowering locus c gene coding region operably linked to a constitutive promoter, wherein the transgenic plant expresses the one or more cell wall degrading enzymes and a transcription factor encoded by the flowering locus c gene that delays flowering while increasing biomass and enabling isolation of increased amounts of the hydrolyzing enzyme from the transgenic plant as compared to a non-transgenic plant from which the transgenic plant is derived. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
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24. A transgenic plant capable of expressing cell wall degrading enzymes comprising:
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(a) at least one DNA encoding a β
-glucosidase as a first cell wall degrading enzyme which is operably linked to a nucleotide sequence encoding a signal peptide directing the β
-glucosidase to a plastid or apoplast of the transgenic plant;
(b) at least one DNA encoding a ligninase as a second cell wall degrading enzyme which is operably linked to a nucleotide sequence encoding a signal peptide directing the ligninase to a plastid or apoplast of the transgenic plant, (c) at least one DNA encoding a xylanase as a third cell wall degrading enzyme which is operably linked to a nucleotide sequence encoding a signal peptide directing the xylanase to a plastid or apoplast of the transgenic plant; and
(d) at least one DNA comprising a flowering locus c gene coding region operably linked to a constitutive promoter, wherein the transgenic plant expresses the cell wall degrading enzymes and a transcription factor encoded by the flowering locus c gene that delays flowering while increasing biomass and enabling isolation of increased amounts of the cell wall degrading enzymes from the transgenic plant as compared to a non-transgenic plant from which the transgenic plant is derived. - View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32, 33)
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34. A method for making an enzyme extract comprising one or more cell wall degrading enzymes comprising:
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(a) providing a transgenic plant capable of expressing one or more cell wall degrading enzymes comprising at least one DNA comprising a cell wall degrading enzyme coding region operably linked to a nucleotide sequence encoding a signal peptide directing the cell wall degrading enzyme encoded by the DNA to a plastid or apoplast of the transgenic plant; and
at least one DNA comprising a flowering locus c gene coding region operably linked to a constitutive promoter, wherein the transgenic plant expresses the one or more cell wall degrading enzymes and a transcription factor encoded by the flowering locus c gene that delays flowering while increasing biomass and enabling isolation of increased amounts of the hydrolyzing enzyme from the transgenic plant as compared to a non-transgenic plant from which the transgenic plant is derived;
(b) growing the transgenic plant for a time to accumulate the one or more cell wall degrading enzymes;
(c) harvesting the transgenic plant which has accumulated the one or more cell wall degrading enzymes; and
(d) grinding the transgenic plant to provide an enzyme extract comprising the one or more cell wall degrading enzymes that accumulated in the transgenic plant.
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35. A method for converting lignocellulosic material to fermentable sugars comprising:
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(a) providing a transgenic plant capable of expressing one or more cell wall degrading enzymes comprising at least one DNA comprising a cell wall degrading enzyme coding region operably linked to a nucleotide sequence encoding a signal peptide directing the cell wall degrading enzyme encoded by the DNA to a plastid or apoplast of the transgenic plant; and
at least one DNA comprising a flowering locus c gene coding region operably linked to a constitutive promoter, wherein the transgenic plant expresses the one or more cell wall degrading enzymes and a transcription factor encoded by the flowering locus c gene that delays flowering while increasing biomass and enabling isolation of increased amounts of the hydrolyzing enzyme from the transgenic plant as compared to a non-transgenic plant from which the transgenic plant is derived;
(b) growing the transgenic plant for a time sufficient for the transgenic plant to accumulate the one or more cell wall degrading enzymes;
(c) harvesting the transgenic plant which has accumulated the one or more cell wall degrading enzymes;
(d) grinding the transgenic plant to provide an enzyme extract comprising the one or more cell wall degrading enzymes that accumulated in the transgenic plant;
(e) incubating the lignocellulosic material in the enzyme extract to produce the fermentable sugars from the lignocellulose in the plant material; and
(f) extracting the fermentable sugars produced from the lignocellulosic material.
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Specification