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. A nucleic acid construct comprising an isolated polynucleotide encoding a polypeptide having lipase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct the production of the polynucleotide in an expression host and wherein the polynucleotide is 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: 6;
(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: 5 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: 5.
2. The nucleic acid construct of claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 90% identity with the mature polypeptide of SEQ ID NO: 6.
3. The nucleic acid construct of claim 2 , wherein the polypeptide comprises an amino acid sequence having at least 95% identity with the mature polypeptide of SEQ ID NO: 6.
4. The nucleic acid construct of claim 3 , wherein the polypeptide comprises an amino acid sequence having at least 97% identity with the mature polypeptide of SEQ ID NO: 6.
5. The nucleic acid construct of claim 1 , wherein the polypeptide comprises or consists of the amino acid sequence of SEQ ID NO: 6 or the mature polypeptide thereof.
6. The nucleic acid construct of claim 1 , wherein the polynucleotide comprises a nucleotide sequence which hybridizes under at least high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 5 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.
7. The nucleic acid construct of claim 6 , wherein the polynucleotide comprises a nucleotide sequence which hybridizes under at least very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 5 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.
8. The nucleic acid construct of claim 1 , wherein the polynucleotide comprises a nucleotide sequence having at least 90% identity with the mature polypeptide coding sequence of SEQ ID NO: 5.
9. The nucleic acid construct of claim 8 , wherein the polynucleotide comprises a nucleotide sequence having at least 95% identity with the mature polypeptide coding sequence of SEQ ID NO: 5.
10. The nucleic acid construct of claim 9 , wherein the polynucleotide comprises a nucleotide sequence having at least 97% identity with the mature polypeptide coding sequence of SEQ ID NO: 5.
11. The nucleic acid construct of claim 1 , wherein the polynucleotide comprises or consists of the nucleotide sequence of SEQ ID NO: 5 or the mature polypeptide coding sequence thereof.
12. The nucleic acid construct of claim 1 , wherein the polynucleotide is encoded by the polynucleotide contained in plasmid pJLin175 which is contained in E. coli NRRL B-30783.
13. The nucleic acid construct of claim 1 , wherein the mature polypeptide is amino acids 21 to 598 of SEQ ID NO: 6.
14. The nucleic acid construct of claim 1 , wherein the mature polypeptide coding sequence is nucleotides 61 to 1922 of SEQ ID NO: 5.
15. An expression vector comprising the nucleic acid construct of claim 1 .
16. A recombinant host cell comprising the nucleic acid construct of claim 1 operably linked to one or more control sequences that direct the production of the polypeptide.
17. A method for producing a polypeptide having lipase activity, comprising: (a) cultivating the host cell of claim 16 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.
18. A method for producing a polypeptide having lipase activity, comprising: (a) cultivating a plant cell comprising the nucleic acid construct of claim 1 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.
19. The method of claim 18 , wherein the polypeptide comprises an amino acid sequence having at least 90% identity with the mature polypeptide of SEQ ID NO: 6.
20. The method of claim 18 , wherein the polypeptide comprises an amino acid sequence having at least 95% identity with the mature polypeptide of SEQ ID NO: 6.
21. The method of claim 18 , wherein the polypeptide comprises an amino acid sequence having at least 97% identity with the mature polypeptide of SEQ ID NO: 6.
22. The method of claim 18 , wherein the polypeptide comprises or consists of the amino acid sequence of SEQ ID NO: 6 or the mature polypeptide thereof.
23. The method of claim 18 , wherein the nucleotide sequence hybridizes under at least high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 5 or its full-length complementary strand, wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5X 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 2X SSC, 0.2% SDS at 65° C.
24. The method of claim 18 , wherein the nucleotide sequence hybridizes under at least very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 5 or its full-length complementary strand, wherein very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5X 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 2X SSC, 0.2% SDS at 70° C.
25. The method of claim 18 , wherein the nucleotide sequence has at least 90% identity with the mature polypeptide coding sequence of SEQ ID NO: 5.
26. The method of claim 18 , wherein the nucleotide sequence has at least 95% identity with the mature polypeptide coding sequence of SEQ ID NO: 5.
27. The method of claim 18 , wherein the nucleotide sequence has at least 97% identity with the mature polypeptide coding sequence of SEQ ID NO: 5.
28. The method of claim 18 , wherein the nucleotide sequence comprises or consists of SEQ ID NO: 5 or the mature polypeptide coding sequence thereof.
29. The method of claim 18 , wherein the polynucleotide is encoded by the polynucleotide contained in plasmid pJLin175 which is contained in E. coli NRRL B-30783.
30. The method of claim 18 , wherein the mature polypeptide is amino acids 21 to 598 of SEQ ID NO: 6.
31. The method of claim 18 , wherein the mature polypeptide coding sequence of the polynucleotide is nucleotides 61 to 1922 of SEQ ID NO: 5.