Enhanced protein expression in <i>Bacillus</i>

摘要

The present invention provides cells that have been genetically manipulated to have an altered capacity to produce expressed proteins. In particular, the present invention relates to Gram-positive microorganisms, such as Bacillus species having enhanced expression of a protein of interest, wherein one or more chromosomal genes have been inactivated, and preferably wherein one or more chromosomal genes have been deleted from the Bacillus chromosome. In some further embodiments, one or more indigenous chromosomal regions have been deleted from a corresponding wild-type Bacillus host chromosome.

主权项

1. A method for enhancing the expression of a protein of interest in Bacillus comprising:
(a) introducing a DNA construct including a selective marker and an inactivating chromosomal segment into a Bacillus host strain, wherein said DNA construct is integrated into the chromosome of said Bacillus host strain, resulting in the deletion of an indigenous chromosomal region or fragment thereof from said Bacillus host cell to produce an altered Bacillus strain, wherein said indigenous chromosomal region is the PPS region; and (b) growing the altered Bacillus strain under suitable conditions, wherein expression of a protein of interest is greater in the altered Bacillus strain compared to the expression of the protein of interest in a Bacillus host cell that has not been altered.