Serine proteases of bacillus species

Abstract

The present disclosure relates to serine proteases cloned from Bacillus akibai and Bacillus clarkii , and variants thereof. Compositions containing the serine proteases are suitable for use in cleaning fabrics and hard surfaces, as well as in a variety of industrial applications.

1 . A recombinant polypeptide of a  Bacillus akibai/clarkii -clade subtilisin, or an active fragment thereof, wherein the recombinant polypeptide or the active fragment thereof has proteolytic activity. 
     
     
         2 . The recombinant polypeptide or the active fragment thereof of  claim 1 , wherein the recombinant polypeptide or the active fragment thereof comprises an amino acid sequence of SEQ ID NO:42, 43, 44, 45, 46, 47, or 48. 
     
     
         3 . The recombinant polypeptide or the active fragment thereof of  claim 2 , further comprising an amino acid sequence having at least 70% or 72% identity to an amino acid sequence of SEQ ID NO:3, 6, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41 or 84. 
     
     
         4 - 6 . (canceled) 
     
     
         7 . The recombinant polypeptide of  claim 1 , wherein the polypeptide has protease activity in the presence of a surfactant. 
     
     
         8 . The recombinant polypeptide of  claim 7 , wherein the protease activity comprises casein hydrolysis. 
     
     
         9 . The recombinant polypeptide of  claim 7 , wherein the polypeptide retains at least 50% of its maximal protease activity at a pH range of 8 to 12 and/or a temperature range of 50° C. to 75° C. 
     
     
         10 . (canceled) 
     
     
         11 . The recombinant polypeptide of  claim 1 , wherein the polypeptide has cleaning activity in a detergent composition. 
     
     
         12 . The recombinant polypeptide of  claim 11 , wherein the detergent composition is an automatic dish washing detergent. 
     
     
         13 . The recombinant polypeptide of  claim 12 , wherein the cleaning activity comprises hydrolysis of an egg yolk substrate. 
     
     
         14 . The recombinant polypeptide of  claim 11 , wherein the detergent composition is a laundry detergent. 
     
     
         15 - 16 . (canceled) 
     
     
         17 . The recombinant polypeptide of  claim 14 , wherein the cleaning activity comprises hydrolysis of a substrate selected from the group consisting of blood, milk, ink and combinations thereof. 
     
     
         18 . A composition comprising a surfactant and the recombinant polypeptide of  claim 1 . 
     
     
         19 .- 21 . (canceled) 
     
     
         22 . The composition of  claim 18 , wherein the composition is a detergent composition. 
     
     
         23 . The composition of  claim 22 , wherein the detergent composition is selected from the group consisting of a laundry detergent, a fabric softening detergent, a dishwashing detergent, and a hard-surface cleaning detergent. 
     
     
         24 . The composition of  claim 18 , wherein said composition further comprises at least one calcium ion and/or zinc ion, at least one stabilizer, from about 0.001% to about 1.0 weight % of said recombinant polypeptide, at least one bleaching agent, at least one adjunct and/or one or more additional enzymes or enzyme derivatives selected from the group consisting of acyl transferases, alpha-amylases, beta-amylases, alpha-galactosidases, arabinosidases, aryl esterases, beta-galactosidases, carrageenases, catalases, cellobiohydrolases, cellulases, chondroitinases, cutinases, endo-beta-1, 4-glucanases, endo-beta-mannanases, esterases, exo-mannanases, galactanases, glucoamylases, hemicellulases, hyaluronidases, keratinases, laccases, lactases, ligninases, lipases, lipoxygenases, mannanases, oxidases, pectate lyases, pectin acetyl esterases, pectinases, pentosanases, peroxidases, phenoloxidases, phosphatases, phospholipases, phytases, polygalacturonases, proteases, pullulanases, reductases, rhamnogalacturonases, beta-glucanases, tannases, transglutaminases, xylan acetyl-esterases, xylanases, xyloglucanases, xylosidases, metalloproteases, additional serine proteases, and combinations thereof. 
     
     
         25 - 27 . (canceled) 
     
     
         28 . The composition of  claim 18 , wherein said composition is phosphate-free or contains phosphate and/or is borate-free or contains borate. 
     
     
         29 - 36 . (canceled) 
     
     
         37 . A method of cleaning comprising contacting a surface or an item with the composition of  claim 18 . 
     
     
         38 - 44 . (canceled) 
     
     
         45 . A method for producing a recombinant polypeptide comprising:
 stably transforming a host cell with an expression vector comprising a polynucleotide encoding the polypeptide of  claim 1 ;   cultivating said transformed host cell under conditions suitable for said host cell to produce said polypeptide; and   recovering said polypeptide.   
     
     
         46 - 50 . (canceled) 
     
     
         51 . A polynucleotide comprising a nucleic acid sequence:
 (i) encoding an amino acid sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:14 and SEQ ID NO:17;   (ii) encoding an amino acid sequence having 70% identity to an amino acid sequence of SEQ ID NO:3, 6, 14, 17, 20, 23, 26, 29, or 84;   (iii) encoding an amino acid sequence having 70% identity to an amino acid sequence of SEQ ID NO:3, 6, 14, or 17;   (iv) encoding an amino acid sequence having 72% identity to an amino acid sequence of SEQ ID NO:3, 6, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41, or 84;   (v) encoding an amino acid sequence having 72% identity to an amino acid sequence of SEQ ID NO:3, 6, 11, 14, or 17;   (vi) having at least 70% identity to SEQ ID NO:1, 4, 12, 15, 18, 21, 24, or 27;   (vii) having at least 70% identity to SEQ ID NO:1, 4, 12, or 15;   (viii) having at least 72% identity to SEQ ID NO:1, 4, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, or 39;   (ix) having at least 72% identity to SEQ ID NO:1, 4, 9, 12, or 15; or   (x) having complementarity to SEQ ID NO: 1, 4, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, or 39.   
     
     
         52 . (canceled) 
     
     
         53 . An expression vector comprising the polynucleotide of  claim 51 . 
     
     
         54 - 56 . (canceled) 
     
     
         57 . A textile, leather, or feather processing composition or a contact lens or wound cleaning composition comprising the polypeptide of  claim 1 . 
     
     
         58 - 61 . (canceled)