Bacillus Host Cell

Abstract

A Bacillus licheniformis mutant host cell derived from a parent B. licheniformis host cell, which mutant host cell is mutated in one or more gene(s) encoding one or more secreted polypeptide(s) which is at least 80% identical to one or more of the polypeptides shown in SEQ ID NO's: 2 to 248, wherein the mutant host cell secretes at least 5% less of the one or more secreted polypeptide(s) than the parent host cell, when they are cultivated under comparable conditions.

1 . An isolated mutant host cell of a  Bacillus licheniformis  host cell, which mutant host cell comprises a mutation in a gene encoding a secreted polypeptide which has an amino acid sequence which is at least 80% identical to SEQ ID NO:166, wherein the mutant host cell secretes at least 5% less of the secreted polypeptide than the parent host cell, when they are cultivated under comparable conditions. 
     
     
         2 . A host cell in accordance with  claim 1 , which is mutated by a partial or complete deletion of the gene encoding the secreted polypeptide. 
     
     
         3 . A host cell in accordance with  claim 1 , which is mutated in two or more genes encoding two or more secreted polypeptides. 
     
     
         4 . A host cell in accordance with  claim 1 , which comprises one or more heterologous gene(s) encoding one or more heterologous polypeptide(s). 
     
     
         5 . A host cell in accordance with  claim 4 , wherein the heterologous gene(s) is present in at least two copies. 
     
     
         6 . A host cell in accordance with  claim 4 , wherein the heterologous gene(s) are stably integrated into the genome of the cell. 
     
     
         7 . A host cell in accordance with  claim 4 , wherein the heterologous gene(s) is integrated into the genome of the cell without leaving any antibiotic resistance marker genes at the site of integration. 
     
     
         8 . A host cell in accordance with  claim 4 , wherein the heterologous gene(s) are transcribed from a heterologous promoter or from an artificial promoter. 
     
     
         9 . A host cell in accordance with  claim 4 , wherein the heterologous gene(s) are comprised in an operon, preferably a polycistronic operon. 
     
     
         10 . A host cell in accordance with  claim 4 , wherein the heterologous polypeptide(s) is an antimicrobial peptide, or a fusion peptide comprising a peptide part which in its native form has antimicrobial activity. 
     
     
         11 . A host cell in accordance with  claim 4 , wherein the heterologous polypeptide(s) has biosynthetic activity and produces a compound or an intermediate of interest. 
     
     
         12 . A host cell in accordance with  claim 11 , wherein the compound or intermediate of interest comprises vitamins, amino acids, antibiotics, carbohydrates, or surfactants. 
     
     
         13 . A host cell in accordance with  claim 12 , wherein the carbohydrates comprise hyaluronic acid. 
     
     
         14 . A host cell in accordance with  claim 4 , wherein the heterologous polypeptide(s) is an enzyme, preferably a secreted enzyme. 
     
     
         15 . A host cell in accordance with  claim 14 , wherein the enzyme is an enzyme of a class selected from the group of enzyme classes consisting of oxidoreductases (EC 1), transferases (EC 2), hydrolases (EC 3), lyases (EC 4), isomerases (EC 5), and ligases (EC 6). 
     
     
         16 . A host cell in accordance with  claim 15 , wherein the enzyme is an enzyme with an activity selected from the group of enzyme activities consisting of aminopeptidase, amylase, amyloglucosidase, mannanase, carbohydrase, carboxypeptidase, catalase, cellulase, chitinase, cutinase, cyclodextrin glycosyltransferase, deoxyribonuclease, esterase, galactosidase, beta-galactosidase, glucoamylase, glucose oxidase, glucosidase, haloperoxidase, hemicellulase, invertase, isomerase, laccase, ligase, lipase, lyase, mannosidase, oxidase, pectinase, peroxidase, phytase, phenoloxidase, polyphenoloxidase, protease, ribonuclease, transferase, transglutaminase, and xylanase. 
     
     
         17 . A host cell in accordance with  claim 16 , wherein the enzyme is an amylase or a mannanase. 
     
     
         18 . A process for producing at least one product of interest in a  Bacillus licheniformis  mutant host cell, comprising cultivating a  B. licheniformis  mutant host cell of  claim 1  in a suitable medium, whereby the said product is produced. 
     
     
         19 . A process in accordance with the process of  claim 18 , further comprising isolating or purifying the product of interest. 
     
     
         20 . A method of using the  Bacillus licheniformis  mutant host cell of  claim 1  comprising:
 producing at least one product of interest comprising cultivating the mutant host cell in a suitable medium whereby the said product is produced.   
     
     
         21 . A method in accordance with  claim 20 , further comprising isolating or purifying the product of interest. 
     
     
         22 . A host cell in accordance with  claim 1 , wherein the amino acid sequence is at least 85% identical to SEQ ID NO:166. 
     
     
         23 . A host cell in accordance with  claim 1 , wherein the amino acid sequence is at least 90% identical to SEQ ID NO:166. 
     
     
         24 . A host cell in accordance with  claim 1 , wherein the amino acid sequence is at least 95% identical to SEQ ID NO:166.