POLYOL OXIDASES
The present invention provides compositions and methods for producing a polyol oxidase in micoroorganisms, and the use of polyol oxidases in cleaning compositions. The invention includes cleaning compositions that contain combinations of two or more POx oxidases, and cleaning compositions that contain combinations of two or more POx oxidases and a perhydrolase. In particular, the invention provides methods for expressing polyol oxidases in bacterial hosts for use in detergent applications for cleaning, bleaching and disinfecting.
1 . An isolated chimeric polynucleotide comprising a sequence encoding a mature polyol oxidase enzyme (POx) comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:4, said sequence being operably linked to a sequence encoding a heterologous secretory signal peptide.
2 . The isolated polynucleotide of claim 1 , wherein said heterologous secretory signal peptide is derived from a prokaryotic microorganism.
3 . (canceled)
4 . The chimeric polynucleotide of claim 2 , wherein said heterologous secretory signal peptide is selected from the signal peptide encoded by the S. coelicolor gene SCO7637, the signal peptide encoded by the S. lividans gene SCO 0624 and the signal peptide encoded by the B. subtilis gene P43379.
5 . A recombinant expression vector comprising the isolated chimeric polynucleotide of claim 1 .
6 . A host cell comprising the recombinant expression vector of claim 5 .
7 . A host cell comprising a chimeric polynucleotide comprising a sequence encoding a mature polyol oxidase enzyme (POx) comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:4, said sequence being operably linked to a sequence encoding a heterologous secretory signal peptide.
8 . The host cell of claim 7 , wherein said POx has sorbitol oxidase and/or xylitol oxidase activity.
9 . The host cell of claim 7 , wherein said chimeric polynucleotide is present in the genome of said host cell or in a vector that autonomously replicates in said host cell.
10 . The host cell of claim 7 , wherein said host cell is an S. lividans , a B. subtilis or an Acidothermus cellulolyticus host cell.
11 . A method for producing a polypeptide having POx activity comprising:
(a) transforming a host cell with a recombinant expression vector, said expression vector comprising a polynucleotide sequence encoding a POx polypeptide comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:4;
(b) growing said host cell comprising said recombinant expression vector under conditions suitable for the expression of said POx polypeptide; and
(c) recovering said POx polypeptide.
12 . The method of claim 11 , wherein said POx activity is sorbitol and/or xylitol activity.
13 . The method of claim 11 , wherein said host cell is selected from Bacillus cells or Streptomyces cells.
14 . The method of claim 11 , wherein said expression of said POx is extracellular.
15 . The method of claim 11 , wherein said expression of said POx is intracellular.
16 - 28 . (canceled)
29 . The method of claim 11 , wherein said polynucleotide sequence encoding said POx polypeptide is operably linked to a sequence encoding a heterologous secretory signal peptide.
30 . The method of claim 29 , wherein said heterologous secretory signal peptide is derived from a prokaryotic microorganism.
31 . The method of claim 30 , wherein said heterologous secretory signal peptide is selected from the signal peptide encoded by the S. coelicolor gene SCO7637, the signal peptide encoded by the S. lividans gene SCO 0624 and the signal peptide encoded by the B. subtilis gene P43379.