Thermal tolerant avicelase from Acidothermus cellulolyticus
View Patent ↗The invention provides a thermal tolerant (thermostable) cellulase, AviIII, that is a member of the glycoside hydrolase (GH) family. AviIII was isolated and characterized from Acidothermus cellulolyticus and, like many cellulases, the disclosed polypeptide and/or its derivatives may be useful for the conversion of biomass into biofuels and chemicals.
1. An isolated polynucleotide molecule encoding a polypeptide comprising the sequence of SEQ ID NO: 1 or a fragment of SEQ ID NO: 1 having cellulase activity.
2. The isolated polynucleotide molecule of claim 1 , further comprising a nucleic acid sequence encoding a heterologous protein in frame with the polynucleotide molecule.
3. The isolated polynucleotide molecule of claim 2 , wherein the heterologous protein is a peptide tag.
4. The isolated polynucleotide molecule of claim 3 , wherein the peptide tag is 6-His, thioredoxin, hemaglutinin, GST, or OmpA signal sequence tag.
5. The isolated polynucleotide molecule of claim 2 , wherein the heterologous protein is capable of binding a carbohydrate molecule.
6. The isolated polynucleotide molecule of claim 1 , operably linked to a transcriptional or translational regulatory sequence.
7. The isolated polynucleotide molecule of claim 6 , wherein the transcriptional or translational regulatory sequence comprises a transcriptional promoter or enhancer.
8. A vector comprising the polynucleotide molecule of claim 7 .
9. A host cell genetically engineered to express a polypeptide from the polynucleotide molecule of claim 7 .
10. The host cell of claim 9 , wherein the host cell is a plant cell.
11. The host cell of claim 9 , wherein the host cell is a fungus.
12. The host cell of claim 9 , wherein the host cell is a bacterial cell.
13. The host cell of claim 9 , wherein the host cell is a yeast.
14. A method for producing the polypeptide encoded by the polynucleotide molecule of claim 7 , the method comprising:
incubating a host cell genetically engineered to express the polynucleotide molecule of claim 7 under conditions that allow the polypeptide to be produced.
15. The method of claim 14 , further comprising the step of:
isolating the polypeptide from the incubated host cells.
16. The method of claim 14 , wherein the host cell is a plant cell.
17. The method of claim 14 , wherein the host cell is a bacterial cell.
18. The method of claim 14 , wherein the host cell is genetically engineered to express a selectable marker.
19. The polynucleotide molecule of claim 1 , wherein the polypeptide comprises the sequence of SEQ ID NO. 1.
20. The polynucleotide molecule of claim 1 , wherein the polypeptide comprises a fragment of SEQ ID NO. 1 having cellulase activity.