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US 6,670,157 B1Granted
Polypeptides involved in the biosynthesis of streptogramins, nucleotide sequences coding for these polypeptides and their use
AssigneeAventis Pharma S.A.
InventorsVeronique Blanc, Francis Blanche, Joel Crouzet, Nathalie Jacques, Patricia Lacroix, Denis Thibaut, Monique Zagorec, Laurent Debussche, Valerie De Crecy-Lagard
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Abstract
The invention concerns methods for preparing pristinamycin IIB and methods for preparing modified pristinamycin IIB.
Claims20
A method for preparing pristinamycin IIB comprising:
- a) specifically introducing a disabling mutation into at least one gene target in a microorganism that produces pristinamycin IIA and pristinamycin IIB, wherein said gene target is selected from the snaA gene (SEQ ID NO:
- 2) and genes that hybridize to the snaA gene (SEQ ID NO:
- 2) of Streptomyces pristinaespiralis in the presence of formamide at 42° C. with washes at 50° and 60° C. in 2×SCC and 0.1% SDS, wherein said gene target encodes a polypeptide that catalyzes the conversion of pristinamycin IIB to pristinamycin IIA;
- (b) selecting cells containing the mutated gene, wherein the cells are blocked in the conversion of pristinamycin IIB to pristinamycin IIA;
- (c) culturing said cells;
- and
- (d) recovering said pristinamycin IIB.
4 more elements
A method for preparing modified pristinamycin IIB comprising:
- a) specifically introducing a disabling mutation into at least one gene target in a microorganism that produces pristinamycin IIA and pristinamycin IIB, wherein said gene target is selected from the snaA gene (SEQ ID NO:
- 2) and genes that hybridize to the snaA gene (SEQ ID NO:
- 2) of Streptomyces pristinaespiralis in the presence of formamide at 42° C. with washes at 50° and 60° C. in 2×SCC and 0.1% SDS, wherein said gene target encodes a polypeptide that catalyzes the conversion of pristinamycin IIB to pristinamycin IIA;
- (b) selecting cells containing the mutated gene, wherein the cells are blocked in the conversion of pristinamycin IIB to pristinamycin IIA;
- (c) culturing said cells;
- and
- (d) recovering said pristinamycin IIB;
- and
- (e) modifying the pristinamycin IIB recovered from the cells.
6 more elements
Dependencies are inferred from the claim text.