Gist-Brocades N.v.

Dsm N.v.

Genencor International, Inc.

Christiaan A Van Eekelen

Johannes C Van Der Laan

Leo J Mulleners

Onno Misset

Roelck A Cuperus

Leonardus J Mulleners

Johan H Lensink

Transformed prokaryotic hosts are provided comprising two or more copies of a DNA sequence stably maintained in their chromosome, said DNA sequence comprising a gene encoding a polypeptide of interest wherein said copies are separated by endogenous chromosomal DNA sequences. Methods are also provided for producing said transformed host strains. The transformed host strains are capable of increased production of the polypeptide of interest compared to host strains which already produce said polypeptide. Preferred host strains are Bacillus novo species PB92 which produces a high-alkaline proteolytic enzyme and B. licheniformis strain T5 which produces a thermostable .alpha.-amylase, and mutants and variants of said strains. Preferred poly-peptide encoding genes are the protease encoding gene obtainable from Bacillus PB92 and the .alpha.-amylase encoding gene obtainable from B. licheniformis strain T5.

Stable gene amplification in chromosomal DNA of prokaryotic

Transformed prokaryotic hosts are provided comprising two or more copies of a DNA sequence stably maintained in their chromosome, said DNA sequence comprising a gene encoding a polypeptide of interest wherein said copies are separated by endogenous chromosomal DNA sequences. Methods are also provided for producing said transformed host strains. The transformed host strains are capable of increased production of the polypeptide of interest compared to host strains which already produce said polypeptide. Preferred host strains are Bacillus novo species PB92 which produces a high-alkaline proteolytic enzyme and B. licheniformis strain T5 which produces a thermostable .alpha.-amylase, and mutants and variants of said strains. Preferred polypeptide encoding genes are the protease encoding gene obtainable from Bacillus PB92 and the .alpha.-amylase encoding gene obtainable from B. licheniformis strain T5.

PB92 serine protease muteins and their use in detergents

New proteolytic enzymes are provided exhibiting improved properties for application in detergents, especially laundry detergents. These enzymes are obtained by expression of a gene encoding a proteolytic enzyme having an amino acid sequence which differs at least in one amino acid from the wild type enzyme. Preferred enzymes are certain mutants derived from the serine protease of Bacillus nov. spec. PB92.

Recombinant genetic means for the production of serine protease muteins

Novel methods and novel industrial Bacillus strains are provided for enhanced production of serine protease. Plasmid constructs comprising the serine protease gene are introduced into a compatible host, generally a Bacillus already overproducing serine protease. Preferred host cells are those from Bacillus novo sp. PB92 and the preferred serine protease gene also originates from Bacillus PB92. Integration and maintenance of the plasmid in the host cell chromosome can be achieved by including a temperature sensitive origin of replication in the plasmid construct and growing under selective conditions.

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Bergschenhoek, Netherlands

Christiaan A Van Eekelen