Claims
- 1. A method of increasing secretion of a polypeptide in a cell wherein said cell expresses at least one peptide transport protein comprising:
inactivating at least one peptide transport protein in said cell; and culturing said cell under conditions suitable for expression and secretion of said polypeptide.
- 2. The method of claim 1 wherein said cell is a plant cell.
- 3. The method of claim 1 wherein said cell is a fungal cell.
- 4. The method of claim 1 wherein said cell is a gram-negative microorganism.
- 5. The method of claim 1 wherein said cell is a gram-negative microorganism and is a member of the family Escherichia.
- 6. The method of claim 1 wherein said cell is a gram-positive microorganism.
- 7. The method of claim 1 wherein said cell is a gram-positive microorganism and is a member of the family Bacillus.
- 8. The method of claim 1 wherein said cell is a gram-positive microorganism and is a member of the family Bacillus wherein said member of the family Bacillus is selected from the group consisting of B. licheniformis, B. lentus, B. brevis, B. stearothermophilus, B. alkalophilus, B. amyloliquefaciens, B. coagulans, B. circulans, B. lautus, B. methanolicus, B. anthracis and B. thuringiensis.
- 9. The method of claim 1 wherein said polypeptide is selected from the group consisting of hormone, enzyme, growth factor and cytokine.
- 10. The method of claim 1 wherein said polypeptide is heterologous.
- 11. The method of claim 1 wherein said polypeptide is an enzyme selected from the group consisting of proteases, carbohydrases, reductases, lipases, isomerases, transferases, kinases phophatases, cellulase, endo-glucosidase H, oxidase, alpha-amylase, glucoamylase, lignocellulose hemicellulase, pectinase and ligninase.
- 12. The method of claim 1 wherein said polypeptide is a bacillus protease.
- 13. The method of claim 1 wherein said polypeptide is subtilisin.
- 14. The method of claim 1 wherein said polypeptide is an amylase.
- 15. The method of claim 1 wherein said polypeptide is a bacillus amylase.
- 16. The method of claim 1 wherein said peptide transport protein which is inactivated is a gene product of a dciA operon.
- 17. The method of claim 1 wherein said peptide transport protein which is inactivated is dciAE.
- 18. The method of claim 1 wherein said inactivating comprises mutating a nucleic acid comprising nucleic acid encoding said peptide transport protein.
- 19. The method of claim 18 wherein said mutating causes a frameshift.
- 20. The method of claim 18 wherein said mutating is deleting one or more nucleotides.
- 21. A method for producing a polypeptide in a cell comprising the steps of:
a) obtaining a cell comprising nucleic acid encoding a polypeptide to be produced, said cell further comprising a peptide transport operon wherein at least one gene product of said operon is inactive in said cell; and b) culturing said cell under conditions suitable for expression such that said polypeptide is produced.
- 22. The method of claim 21 wherein said polypeptide is heterologous.
- 23. The method of claim 21 wherein said cell is a gram-positive microorganism and is a member of the family Bacillus.
- 24. The method of claim 21 wherein said cell is a gram-negative microorganism.
- 25. The method of claim 21 wherein said cell is a gram-negative microorganism and is a member of the family Escherichia.
- 24. The method of claim 21 wherein said operon is a dciA operon.
- 25. The method of claim 21 wherein said gene product is inactive as the result of a mutation in said operon.
- 26. The method of claim 25 wherein said mutation is a frameshift mutation.
- 27. The method of claim 25 wherein said mutation is a deletion of one or more nucleotides.
- 28. The method of claim 21 wherein said inactive gene product is dciAE or dciAA.
- 29. A cell comprising a peptide transport operon, wherein said operon has been mutated such that said cell has increased polypeptide secretion.
- 30. The cell of claim 29 wherein said operon is a dciA operon.
- 31. The cell of claim 29 wherein said operon has been mutated to inactivate a gene product of a dciAE gene or a dciAA gene.
- 32. The cell of claim 29 wherein said cell is a gram-positive microorganism.
- 33. The cell of claim 29 wherein said cell is a gram-positive microorganism and is a member of the family Bacillus.
- 34. The cell of claim 29 wherein said cell is a gram-negative microorganism.
- 35. The cell of claim 29 wherein said cell is a gram-negative microorganism and is a member of the family Escherichia.
Priority Claims (1)
Number |
Date |
Country |
Kind |
PCT/US99/31010 |
Dec 1999 |
US |
|
RELATED APPLICATIONS
[0001] The present application is related to PCT/US99/31010 filed Dec. 21, 1999, incorporated herein in its entirety.
Divisions (1)
|
Number |
Date |
Country |
Parent |
09595386 |
Jun 2000 |
US |
Child |
09993525 |
Nov 2001 |
US |