Cellulase-free enzyme compositions and host cells for producing the same
The present invention provides recombinant bacterial cells for producing a detergent-additive protein. In some embodiments, the cells are of the genus Bacillus . In additional embodiments, the cells comprise a genome comprising an inactivated bglC gene, as well as a recombinant nucleic acid for production of at least one secreted detergent-additive protein. In some preferred embodiments, the secreted detergent-additive protein is a protease. The present invention also provides methods of using the bacterial cells to produce at least one detergent-additive protein, as well as cellulase-free compositions containing at least one detergent-additive protein.
1. A recombinant Bacillus sp. host cell comprising a genome comprising an inactivated bglC gene, wherein said cell further comprises a recombinant nucleic acid for production of a secreted detergent-additive protein.
2. The recombinant Bacillus sp. host cell of claim 1 , wherein said inactivated bglC gene contains a deletion, an insertion, a substitution or a rearrangement.
3. The recombinant Bacillus sp. host cell of claim 1 , wherein said Bacillus sp. cell is a B. licheniformis, B. subtilis, B. clausii, B. alkalophilus or B. halodurans cell.
4. The recombinant Bacillus sp. host cell of claim 1 , wherein said secreted detergent-additive protein is an enzyme selected from a protease, an amylase, a pectate lyase, an acyltransferase, an arylesterase and a lipase.
5. The recombinant Bacillus sp. host cell of claim 4 , wherein said enzyme is a protease.
6. The recombinant Bacillus sp. host cell of claim 5 , wherein said protease is a subtilisin.
7. The recombinant Bacillus sp. host cell of claim 1 , wherein said bglC gene encodes a polypeptide that is at least 80% identical to SEQ ID NO:2.
8. A culture of cells comprising:
culture medium; and
a recombinant Bacillus sp. host cell of claim 1 .
9. A method, comprising:
maintaining the culture of cells of claim 8 under conditions suitable to produce said secreted detergent-additive protein.
10. The method of claim 9 , further comprising:
recovering said secreted detergent-additive protein from said culture medium.
11. The method of claim 10 , further comprising combining said secreted detergent-additive protein with a laundry detergent.
12. The method of claim 9 , wherein said secreted detergent-additive protein is a subtilisin protease.
13. A cellulase-free protein composition produced by the method of claim 9 .
14. The cellulase-free protein composition of claim 13 , wherein said composition comprises a secreted detergent-additive protein produced by a Bacillus sp. and is characterized in that the composition is does not have detectable cellulase activity.
15. A cellulase-free laundry detergent comprising a secreted detergent-additive protein produced by the method of claim 9 .
16. The cellulase-free laundry detergent of claim 15 , wherein said detergent comprises a cellulosic polymer.
17. A method of making a cell, comprising:
a) introducing a first recombinant nucleic acid into a Bacillus sp. cell so that said recombinant nucleic acid recombines with the bglC gene of said cell; and
b) introducing a second recombinant nucleic acid into said cell that provides for expression of said secreted detergent-additive protein.
18. The method of claim 17 wherein said nucleic acid inserts into said bglC gene.
19. The method of claim 17 , wherein said nucleic acid deletes at least a portion of said bglC gene.
20. The method of claim 17 , wherein said secreted detergent-additive protein is a subtilisin.