Identification and characterization of the spinactin biosysnthesis gene cluster from spinosyn producing saccharopolyspora spinosa
AssigneeDow AgroSciences LLC
InventorsNadaraj Palaniappan, Paul Lewer, Paul Richard Graupner
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Abstract
This disclosure concerns methods for producing a spinosyn producing strain that comprises modifying a nucleic acid molecule encoding for spinactin by introducing, mutating, deleting, replacing or inactivating a nucleic acid sequence encoding one or more activities encoded by said nucleic acid molecule. Methods for producing a modified Saccharopolyspora spinosa organism are also disclosed.
Claims32
A method for producing a spinosyn producing strain comprising modifying a nucleic acid molecule encoding for spinactin by introducing, mutating, deleting, replacing or inactivating a nucleic acid sequence encoding one or more activities encoded by said nucleic acid molecule, wherein such introduced, mutated, deleted, replaced or inactivated sequence results in a nucleic acid molecule encoding a spinosyn that synthesizes a polyketide other than the polyketide synthesized from a native spinosyn producing strain.
A method for producing a modified Saccharopolyspora spinosa organism, the method comprising:
- providing a nucleic acid comprising a nucleotide sequence within the spinactin biosynthetic gene cluster of S. spinosa;
- mutating the nucleotide sequence;
- and
- introducing the nucleic acid comprising the mutated nucleotide sequence into a host S. spinosa organism, such that the nucleic acid is stably integrated into the genomic DNA of the host S. spinosa organism,
- thereby producing a modified S. spinosa organism, wherein a gene within the spinactin biosynthetic gene cluster no longer expresses a functional protein.
A method for producing a mixture of spinosyns, the method comprising:
- providing a culture of at least one Saccharopolyspora spinosa organism comprising a means for disrupting spinactin synthesis;
- culturing the at least one S. spinosa organism under fen ientation conditions;
- and
- obtaining a mixture of spinosyns from the S. spinosa fermentation culture.
A modified Saccharopolyspora spinosa organism comprising a means for disrupting spinactin biosynthesis, wherein the organism is capable of producing spinosyn A and/or spinosyn D without producing a significant amount of spinactin.
A compound having the formula (I):
- or a salt or N-oxide thereof.
An isolated polynucleotide comprising a nucleotide sequence having at least 80% identity to SEQ ID NO:2, a nucleotide sequence capable of hybridizing to SEQ ID NO:2, complements thereof, and reverse complements thereof.
Dependencies are inferred from the claim text.