Polypeptides having lipase activity and polynucleotides encoding same
View Patent ↗The present invention relates to isolated polypeptides having lipase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods for producing and using the polypeptides.
1. An isolated polynucleotide encoding a polypeptide having lipase activity, selected from the group consisting of:
(a) a polynucleotide encoding a polypeptide having lipase activity comprising an amino acid sequence having at least 90% identity with the mature polypeptide of SEQ ID NO: 2;
(b) a polynucleotide encoding a polypeptide having lipase activity comprising a nucleotide sequence which hybridizes under at least high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 1 or its full-length complementary strand, wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide and a wash three times, each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; and
(c) a polynucleotide encoding a polypeptide having lipase activity comprising a nucleotide sequence having at least 90% identity with the mature polypeptide coding sequence of SEQ ID NO: 1.
2. The isolated polynucleotide of claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 95% identity with the mature polypeptide of SEQ ID NO: 2.
3. The isolated polynucleotide of claim 2 , wherein the polypeptide comprises an amino acid sequence having at least 97% identity with the mature polypeptide of SEQ ID NO: 2.
4. The polynucleotide of claim 1 , wherein the polypeptide comprises or consists of the amino acid sequence of SEQ ID NO: 2 or the mature polypeptide thereof.
5. The isolated polynucleotide of claim 1 , which comprises a nucleotide sequence which hybridizes under at least high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 1 or its full-length complementary strand, wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide and a wash three times, each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.
6. The isolated polynucleotide of claim 5 , which comprises a nucleotide sequence which hybridizes under at least very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 1 or its full-length complementary strand, wherein very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide and a wash three times, each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.
7. The polynucleotide of claim 1 , which comprises a nucleotide sequence having at least 95% identity with the mature polypeptide coding sequence of SEQ ID NO: 1.
8. The polynucleotide of claim 7 , which comprises a nucleotide sequence having at least 97% identity with the mature polypeptide coding sequence of SEQ ID NO: 1.
9. The polynucleotide of claim 1 , which comprises or consists of the nucleotide sequence of SEQ ID NO: 1 or the mature polypeptide coding sequence thereof.
10. The polynucleotide of claim 1 , which is encoded by the polynucleotide contained in plasmid pJLin173 which is contained in E. coli NRRL B-30782.
11. The polynucleotide of claim 1 , wherein the mature polypeptide is amino acids 21 to 562 of SEQ ID NO: 2.
12. The polynucleotide of claim 1 , wherein the mature polypeptide coding sequence is nucleotides 61 to 1686 of SEQ ID NO: 1.
13. A nucleic acid construct or expression vector comprising the polynucleotide of claim 1 operably linked to one or more control sequences that direct the production of the polypeptide in an expression host.
14. A recombinant host cell comprising the polynucleotide of claim 1 operably linked to one or more control sequences that direct the production of the polypeptide.
15. A method for producing a polypeptide having lipase activity, comprising: (a) cultivating the host cell of claim 14 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.
16. A method for producing a polypeptide having lipase activity, comprising: (a) cultivating a transgenic plant or a plant cell comprising the polynucleotide of claim 1 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.
17. A transgenic plant, plant part or plant cell, which has been transformed with the polynucleotide of claim 1 .