Consolidated pretreatment and hydrolysis of plant biomass expressing cell wall degrading enzymes
Methods for consolidated pretreatment and hydrolysis of genetically engineered plants expressing cell wall degrading enzymes are provided. Expression cassettes and vectors for making transgenic plants are described. Plants engineered to express one or more cell wall degrading enzymes using expression cassettes and vectors of the invention are also provided.
1. An engineered plant including a first polynucleotide sequence encoding an amino acid sequence with at least 90% identity to a first reference sequence selected from the group consisting of: SEQ ID NO: 7 and SEQ ID NO: 8.
2. The engineered plant of claim 1 further including a second polynucleotide sequence encoding an amino acid sequence with at least 90% identity to a second reference sequence selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID: 12, wherein the SEQ ID NO selected as the second reference sequence is different than the SEQ ID NO selected as the first reference sequence.
3. The engineered plant of claim 2 further comprising a third polynucleotide sequence encoding an amino acid sequence with at least 90% identity to a third reference sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID: 12 , wherein the SEQ ID NO selected as the third reference sequence is different than the SEQ ID NO selected as the first reference sequence and different than the SEQ ID NO selected as the second reference sequence.
4. The engineered plant of claim 1 , wherein the first polynucleotide sequence, the second polynucleotide sequence, or the third polynucleotide sequence further includes a targeting polynucleotide sequence encoding a targeting peptide selected from the group consisting of: an amyloplast targeting signal, a cell wall targeting peptide, a mitochondrial targeting peptide, a cytosol localization signal, a chloroplast targeting signal, a nuclear targeting peptide, and a vacuole targeting peptide.
5. The engineered plant of claim 4 , wherein the targeting peptide is selected from the group consisting of: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24.
6. The engineered plant of claim 1 , wherein the engineered plant is selected from the group consisting of: corn, sugar cane, sugar beet, sorghum, switchgrass, miscanthus, eucalyptus , willow and poplar.
7. The engineered plant of claim 1 , wherein the first polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the first reference sequence of SEQ ID NO: 7.
8. The engineered plant of claim 2 , wherein the first polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the first reference sequence of SEQ ID NO: 7 and the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 8.
9. The engineered plant of claim 1 , wherein the first polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the first reference sequence of SEQ ID NO: 8.
10. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 1.
11. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 2.
12. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 3.
13. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 4.
14. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 5.
15. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 6.
16. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 9.
17. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 10.
18. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 11.
19. The engineered plant of claim 2 , wherein the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID: 12.
20. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 1.
21. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 2.
22. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 3.
23. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 4.
24. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 5.
25. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 6.
26. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 7.
27. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 8.
28. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 9.
29. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 10.
30. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID NO: 11.
31. The engineered plant of claim 3 , wherein the third polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the third reference sequence of SEQ ID: 12.
32. The engineered plant of claim 3 , wherein the first polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the first reference sequence of SEQ ID NO: 7, the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 4, and the third polynucleotide sequence encodes the amino acid with at least 90% identity to the third reference sequence of SEQ ID NO: 5.
33. The engineered plant of claim 32 , wherein the first polynucleotide sequence encodes the amino acid of SEQ ID NO: 7, the second polynucleotide sequence encodes the amino acid sequence of SEQ ID NO: 4, and the third polynucleotide sequence encodes the amino acid of SEQ ID NO: 5.
34. The engineered plant of claim 3 , wherein the first polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the first reference sequence of SEQ ID NO: 8, the second polynucleotide sequence encodes the amino acid sequence with at least 90% identity to the second reference sequence of SEQ ID NO: 4, and the third polynucleotide sequence encodes the amino acid with at least 90% identity to the third reference sequence of SEQ ID NO: 5.
35. The engineered plant of claim 34 , wherein the first polynucleotide sequence encodes the amino acid of SEQ ID NO: 8, the second polynucleotide sequence encodes the amino acid sequence of SEQ ID NO: 4, and the third polynucleotide sequence encodes the amino acid of SEQ ID NO: 5.