Thermal tolerant exoglucanase from
View Patent ↗The invention provides a thermal tolerant cellulase that is a member of the glycoside hydrolase family. The invention further discloses this cellulase as Gux1. Gux1 has been isolated and characterized from Acidothermus cellulolyticus . The invention further provides recombinant forms of the identified Gux1. Methods of making and using Gux1 polypeptides, including fusions, variants, and derivatives, are also disclosed.
1. A composition comprising an isolated protein with catalytic activity of an exoglucanase, said protein comprising a glycoside hydrolase 48 (GH48) catalytic domain of SEQ ID NO:5, a carbohydrate binding domain (CBD) type III of SEQ ID NO:4 and possessing carbohydrate binding activity, and a carbohydrate binding domain (CBD) type II of SEQ ID NO:7 and possessing carbohydrate binding activity.
2. The composition of claim 1 wherein the protein is further defined as comprising a linker and a signal peptide.
3. An isolated protein with catalytic activity of an exoglucanase having the sequence of SEQ ID NO: 1.
4. The protein of claim 3 further defined as being encoded by a polynucleotide having the sequence of SEQ ID NO: 2.
5. The composition of claim 1 wherein the protein further comprises a linker of SEQ ID NO: 6.
6. The composition of claim 1 , wherein the protein further comprises a peptide tag.
7. The composition of claim 6 wherein the peptide tag is 6-His (SEQ ID NO: 8), thioredoxin, hemaglutinin, GST, or OmpA signal sequence tag.
8. The composition of claim 1 wherein the protein further comprises a substrate targeting moiety.
9. The composition of claim 1 further comprising a carrier.
10. The composition of claim 1 wherein the protein is further defined as comprising a heterologous peptide or protein.
11. The composition of claim 10 wherein the heterologous peptide or protein comprises an immunoglobulin.
12. The composition of claim 10 wherein the heterologous peptide comprises a histidine tag.
13. The composition of claim 10 wherein the heterologous peptide comprises a leucine zipper.
14. The composition of claim 1 further defined as comprising, in combination, the sequence of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 7 in that particular order.
15. The composition of claim 10 wherein the heterologous peptide comprises a fusion protein.
16. The composition of claim 15 , wherein the heterologous peptide is a substrate targeting moiety.
17. The composition of claim 15 , wherein the heterologous peptide is a peptide tag.
18. The composition of claim 17 , wherein the peptide tag is 6-His (SEQ ID NO:8), thioredoxin, hemaglutinin, GST, or OmpA signal sequence tag.
19. The composition of claim 15 , wherein the heterologous peptide is an agent that promotes polypeptide oligomerization.