Transgenic plants containing ligninase and cellulase which degrade lignin and cellulose to fermentable sugars
First Claim
1. A herbaceous transgenic plant which degrades lignocellulose when the transgenic plant is ground to produce a plant material comprising:
- (a) the DNA of SEQ ID NO;
4 encoding a cellulase which is operably linked a nucleotide sequence encoding a signal peptide that directs the cellulase to a plastid or apoplast of the transgenic plant; and
(b) the DNA of SEQ ID NO;
11 encoding a ligninase comprising a lignin peroxidase which is operably linked to a nucleotide sequence encoding a signal peptide that directs the ligninase to the plastid or apoplast of the transgenic plant,wherein the transgenic plant degrades the lignocellulose when ground to produce the plant material.
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Abstract
The present invention provides transgenic plants which after harvest degrade the lignin and cellulose therein to fermentable sugars which can further be fermented to ethanol or other products. In particular, the transgenic plants comprise ligninase and cellulase genes from microbes operably linked to a DNA encoding a signal peptide which targets the fusion polypeptide produced therefrom to an organelle of the plant, in particular the chloroplasts. When the transgenic plants are harvested, the plants are ground to release the ligninase and cellulase which then degrade the lignin and cellulose of the transgenic plants to produce the fermentable sugars.
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Citations
38 Claims
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1. A herbaceous transgenic plant which degrades lignocellulose when the transgenic plant is ground to produce a plant material comprising:
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(a) the DNA of SEQ ID NO;
4 encoding a cellulase which is operably linked a nucleotide sequence encoding a signal peptide that directs the cellulase to a plastid or apoplast of the transgenic plant; and(b) the DNA of SEQ ID NO;
11 encoding a ligninase comprising a lignin peroxidase which is operably linked to a nucleotide sequence encoding a signal peptide that directs the ligninase to the plastid or apoplast of the transgenic plant,wherein the transgenic plant degrades the lignocellulose when ground to produce the plant material. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 36)
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12. A method for producing a herbaceous transgenic plant which degrades lignocellulose when the transgenic plant is ground to produce a plant material comprising:
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(a) providing a first transgenic plant which includes the DNA of SEQ ID NO;
4 encoding a cellulase, which is operably linked to a nucleotide sequence encoding a signal peptide that directs the cellulose to a plastid or apoplast of the transgenic plant, and a second transgenic plant which includes the DNA of SEQ ID NO;
11 encoding a ligninase comprising a lignin peroxidase which is operably linked to a nucleotide sequence encoding a signal peptide that directs the ligninase to the plastid or apoplast of the transgenic plant; and(b) mating by sexual fertilization the first and the second transgenic plants to produce a third transgenic plant which includes the DNA encoding the cellulase and the DNA encoding the ligninase, wherein the transgenic plant which includes the DNA encoding the cellulase and the DNA encoding the ligninase degrades the lignocellulose when ground to produce the plant material. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 37)
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24. A method for converting lignocellulose in a herbaceous transgenic plant material to fermentable sugars comprising:
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(a) providing a herbaceous transgenic plant which includes the DNA of SEQ ID NO;
4 encoding a cellulase, which is operably linked to a nucleotide sequence encoding a signal peptide that directs the cellulase to a plastid or apoplast of the transgenic plant and the DNA of SEQ ID NO;
11 encoding a ligninase comprising a lignin peroxidase which is operably linked to a nucleotide sequence encoding a signal peptide wherein the signal peptide directs the ligninase to the plastid or apoplast of the transgenic plant;(b) growing the transgenic plant for a time sufficient for the transgenic plant to accumulate a sufficient amount of the cellulase and the ligninase in the plastid or apoplast of the transgenic plant; (c) harvesting the transgenic plant which has accumulated the cellulase and ligninase in the plastid or apoplast of the transgenic plant; (d) grinding the transgenic plant for a time sufficient to produce the herbaceous transgenic plant material wherein the cellulase and ligninase produced by the transgenic plant are released from the plastid or apoplast of the transgenic plant; (e) incubating the herbaceous transgenic plant material for a time sufficient for the cellulase and ligninase in the plant material to produce the fermentable sugars from the lignocellulose in the herbaceous transgenic plant material; and (f) extracting the fermentable sugars produced by the cellulase and the ligninase from the lignocellulose in the herbaceous transgenic plant material. - View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 38)
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Specification