Genes and proteins involved in the biosynthesis of lipopeptides
AssigneeEcopia BioSciences, Inc.
InventorsChris M. Farnet, Alfredo Staffa, Emmanuel Zazopoulos
Life of this patent
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Abstract
Genes and proteins involved in the biosynthesis of lipopeptides by microorganisms, in particular the nucleic acids forming the biosynthetic locus for the A54145 lipopeptide from Streptomyces fradiae and the A54145-like lipopeptide from Streptomyces refuineus. These nucleic acids can be used to make expression constructs and transformed host cells for the production of lipopeptides. The genes and proteins allow direct manipulation of lipopeptides and related chemical structures via chemical engineering of the proteins involved in the biosynthesis of A54145.
Claims22
An isolated, purified or enriched nucleic acid comprising a nucleic acid sequence selected from the group consisting of:
- a. SEQ ID NOS:
- 1, 6, 17, 34, and coding regions thereof;
- b. a nucleic acid having at least 75% identity to a nucleic acid of (a);
- and
- c. a nucleic acid complementary to a nucleic acid of (a) or (b).
An isolated, purified or enriched nucleic acid that hybridizes under stringent conditions to any one of A541 ORFs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 (SEQ ID NOS:
- 3, 5, 8, 10, 12, 14, 16, 19, 21, 23, 25, 27, 29, 31, 33) and can substitute for the ORF to which it specifically hybridizes to direct the synthesis of an A54145 compound or analogue.
An isolated gene cluster comprising ORFs encoding polypeptides sufficient to direct the synthesis of an A54145 compound or analogue.
An isolated polypeptide comprising a polypeptide sequence selected from any one of:
- (a) a polypeptide of any one of SEQ ID NOS:
- 2, 4, 7, 9, 11, 13, 15, 18, 20, 22, 24, 26, 28, 30, 32;
- and
- (b) a polypeptide which is at least 75% identical in amino acid sequence to a polypeptide of any one of SEQ ID NOS:
- 2, 4, 7, 9, 11, 13, 15, 18, 20, 22, 24, 26, 28, 30, 32.
An isolated or purified antibody capable of specifically binding to a polypeptide having a sequence selected from the group consisting of SEQ ID NOS:
- 2, 4, 7, 9, 11, 13, 15, 18, 20, 22, 24, 26, 28, 30, 32.
A method of making a polypeptide having a sequence selected from the group consisting of SEQ ID NOS:
- 2, 4, 7, 9, 11, 13, 15, 18, 20, 22, 24, 26, 28, 30, 32 comprising introducing a nucleic acid encoding said polypeptide, said nucleic acid being operably linked to a promoter, into a host cell.
A method of making an A54145 compound or analog comprising culturing a Streptomyces fradiae bacterium under conditions allowing for expression of A541 ORFs 1 to 15 (SEQ ID NOS:
- 3, 5, 8, 10, 12, 14, 16, 19, 21, 23, 25, 27, 29, 31, 33) present in the E. coli strains DH10B having accession nos. IDAC 260202-1, 260202-2 and 260202-3.
Dependencies are inferred from the claim text.