LIPOPEPTIDES AND LIPOPEPTIDE SYNTHETASES
First Claim
1. An engineered lipopeptide synthetase polypeptide which is a deletion mutant of a naturally occurring lipopeptide synthetase polypeptide,wherein the naturally occurring polypeptide comprises a first and second peptide synthetase domain, wherein each peptide synthetase domain comprises a condensation domain (C domain), an adenylation domain (A domain), and a thiolation domain (T domain),wherein the engineered polypeptide comprises a deletion of at least a portion of a C domain, a portion of an A domain, or a portion of a T domain, relative to the naturally occurring lipopeptide synthetase polypeptide, and wherein the engineered polypeptide comprises a fatty acid linkage domain.
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Abstract
Novel lipopeptides, and engineered polypeptides useful in synthesizing lipopeptides are provided. Also provided are methods of making lipopeptides using engineered polypeptides, and methods of using lipopeptides, e.g., as insecticidal and/or antimicrobial agents.
6 Citations
31 Claims
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1. An engineered lipopeptide synthetase polypeptide which is a deletion mutant of a naturally occurring lipopeptide synthetase polypeptide,
wherein the naturally occurring polypeptide comprises a first and second peptide synthetase domain, wherein each peptide synthetase domain comprises a condensation domain (C domain), an adenylation domain (A domain), and a thiolation domain (T domain), wherein the engineered polypeptide comprises a deletion of at least a portion of a C domain, a portion of an A domain, or a portion of a T domain, relative to the naturally occurring lipopeptide synthetase polypeptide, and wherein the engineered polypeptide comprises a fatty acid linkage domain.
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10. An engineered lipopeptide synthetase polypeptide comprising:
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a first peptide synthetase domain of a first peptide synthetase polypeptide, and a second peptide synthetase domain of a second peptide synthetase polypeptide, wherein the first and second peptide synthetase domains are covalently linked such that the engineered lipopeptide synthetase polypeptide produces a lipopeptide comprising an amino acid encoded by the first peptide synthetase domain linked to an amino acid encoded by the second peptide synthetase domain. - View Dependent Claims (11, 12)
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13. An engineered lipopeptide synthetase polypeptide comprising:
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a first peptide synthetase domain of a naturally occurring lipopeptide synthetase polypeptide, and a second peptide synthetase domain of a naturally occurring lipopeptide synthetase polypeptide, wherein the second peptide synthetase domain is covalently linked to a thioesterase domain of a peptide synthetase polypeptide. - View Dependent Claims (14, 15, 16, 17)
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18-21. -21. (canceled)
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24. A lipopeptide consisting of the following amino acid sequence:
- L-Glu-D-Leu-L-Val-L-Asp-D-Leu-L-Leu (SEQ ID NO;
______), wherein the lipopeptide comprises a fatty acid moiety on the L-Glu residue. - View Dependent Claims (25)
- L-Glu-D-Leu-L-Val-L-Asp-D-Leu-L-Leu (SEQ ID NO;
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26. A lipopeptide consisting of the following amino acid sequence:
- L-Glu-X-D-Leu-L-Val-L-Asp-D-Leu-L-Leu (SEQ ID NO;
______), wherein X is any amino acid, and wherein the lipopeptide comprises a fatty acid moiety on the L-Glu residue. - View Dependent Claims (27, 28)
- L-Glu-X-D-Leu-L-Val-L-Asp-D-Leu-L-Leu (SEQ ID NO;
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29. A lipopeptide consisting of the following amino acid sequence:
- L-Leu-D-Leu-L-Val-L-Asp-D-Leu-L-Leu (SEQ ID NO;
______), wherein the lipopeptide comprises a fatty acid moiety on the first L-Leu residue. - View Dependent Claims (30)
- L-Leu-D-Leu-L-Val-L-Asp-D-Leu-L-Leu (SEQ ID NO;
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31-34. -34. (canceled)
Specification