PLANTS EXPRESSING CELL WALL DEGRADING ENZYMES AND EXPRESSION VECTORS
Vectors for expression of proteins in plants are described. The proteins may be enzymes and the enzymes can be but are not limited to cell wall degrading enzymes. A number of plants designed to express specific cell wall degrading enzymes are provided. The plants may have industrial and/or agricultural applications. Methods and materials for making the expression vectors and for making the plants are provided. Processes for which the plants could be used in industrial and agricultural applications are also provided.
1 . A transgenic plant including a nucleic acid encoding an amino acid sequence with at least 90% identity to a sequence selected from SEQ ID NOS: 44-115.
2 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 44-45, 49-54, 57-59, 85-86, 94-96, 104-109 and 113-115.
3 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 47 and 55.
4 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 46, 48 and 56.
5 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 60-67, 70 and 75.
6 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 68-69, 71-74, 76-77 and 112.
7 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 78-84.
8 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 97-103.
9 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 87-93 and 110-111.
10 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 44, 45, 49 and 54.
11 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 45, 87, 104-106 and 113.
12 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 90% identity to a sequence selected from SEQ ID NOS: 50-53, 57-59, 94-96, 104-109 and 113-115.
13 . The transgenic plant of claim 1 , wherein the amino acid sequence has at least 95% identity to a sequence selected from SEQ ID NOS: 44-115.
14 . The transgenic plant of claim 1 , wherein the amino acid sequence has 100% identity to a sequence selected from SEQ ID NOS: 44-115.
15 . The transgenic plant of claim 1 , wherein the transgenic plant is one of corn, switchgrass, miscanthus, sugarcane or sorghum.
16 . The transgenic plant of claim 1 , wherein the transgenic plant is corn.
17 . The transgenic plant of claim 1 , wherein the transgenic plant is switchgrass.
18 . The transgenic plant of claim 1 , wherein the transgenic plant is made by agrobacterium mediated transformation using a plasmid having a nucleotide sequence having at least 90% identity to the sequence of one of SEQ ID NOS: 188-283.
19 . The transgenic plant of claim 1 , wherein the transgenic plant is made by agrobacterium mediated transformation using a plasmid having a nucleotide sequence having at least 90% identity to the sequence of SEQ ID NO: 207.
20 . A transgenic plant including a first nucleic acid that is capable of hybridizing under conditions of moderate stringency to a second nucleic acid consisting of a nucleotide sequence selected from SEQ ID NOS: 116-187 or the complement thereof.
21 . The transgenic plant of claim 20 , wherein first nucleic acid is capable of hybridizing under conditions of high stringency to the second nucleic acid consisting of a nucleotide sequence selected SEQ ID NOS: 116-187 or the complement thereof.
22 . The transgenic plant of claim 20 , wherein the transgenic plant is one of corn, switchgrass, miscanthus, sugarcane or sorghum.
23 . The transgenic plant of claim 20 , wherein the transgenic plant is corn.
24 . The transgenic plant of claim 20 , wherein the transgenic plant is switchgrass.
25 . The transgenic plant of claim 20 , wherein the transgenic plant is made by agrobacterium mediated transformation using a plasmid having a first nucleic acid capable of hybridizing under conditions of high stringency to a second nucleic acid consisting of the sequence of one of SEQ ID NOS: 116-187.
26 . The transgenic plant of claim 20 , wherein the transgenic plant is made by agrobacterium mediated transformation using a plasmid having the sequence of one of SEQ ID NOS: 188-283.
27 . A vector including a first nucleic acid capable of hybridizing under conditions of high stringency to a second nucleic acid consisting of the sequence of one of SEQ ID NOS: 116-187.
28 . A vector including a nucleic acid having a sequence with at least 90% identity to a reference sequence selected from SEQ ID NOS: 188-283.
29 . The vector of claim 28 , wherein the nucleic acid has a sequence with at least 90% identity to the reference of SEQ ID NOS: 207.
30 . A method of processing plant biomass comprising:
pretreating a plant or part thereof through mixing the plant or part thereof with liquid to form a mixture having a liquid to solid ratio of less than or equal to 15, and providing conditions to maintain the mixture at a temperature less than or equal to 100° C.; and
providing one or more enzyme for enzymatic hydrolysis of lignocellulosic material.
31 . The method of claim 30 , wherein the plant is a transgenic plant of claim 1 .
32 . The method of claim 30 further comprising providing a transgenic plant or part thereof of claim 1 .
33 . The method of claim 30 , wherein the liquid to solid ratio is selected from a value less than or equal to 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1.
34 . The method of claim 30 , wherein the liquid to solid ratio is 8 or less.
35 . The method of claim 30 , wherein the step of pretreating includes maintaining the temperature of less than or equal to 100° C. for at least four hours.
36 . The method of claim 30 , wherein the temperature of less than or equal to 100° C. is 40° C. to 90° C.
37 . The method of claim 30 , wherein the liquid is water.
38 . The method of claim 30 , wherein the liquid includes water, ammonium bisulfite and ammonium carbonate.
39 . The method of claim 38 , wherein the ammonium bisulfite is at a concentration of 8% to 38% on a wt./wt. basis with the plant or part thereof.
40 . The method of claim 38 , wherein the ammonium carbonate is at a pH of 7.6 to 8.5 and concentration of 4% to 19% on a wt./wt. basis with the plant or part thereof.
41 . The method of claim 30 , wherein the step of providing one or more enzyme includes providing at least one of an endoglucanase, a β-glucosidase and a cellobiohydrolase.
42 . The method of claim 41 , wherein the step of providing one or more enzyme includes providing a xylanase.