Patent classifications
C12P19/62
Fidaxomicin purification method
A fidaxomicin purification method, comprising: fermenting Actinoplanes sp. HS-16-20 to generate fermented liquid; conducting solid/liquid separation on the fermented liquid, soaking mycelium in an organic solvent, and filtering to obtain a solution containing fidaxomicin; treating the solution with nanofiltration concentrate, and separating to obtain fidaxomicin crude product; conducting preparative column chromatography on the fidaxomicin crude product, eluting with an acid aqueous solution containing an organic solvent, and separating to obtain the refined fidaxomicin product.
Fidaxomicin purification method
A fidaxomicin purification method, comprising: fermenting Actinoplanes sp. HS-16-20 to generate fermented liquid; conducting solid/liquid separation on the fermented liquid, soaking mycelium in an organic solvent, and filtering to obtain a solution containing fidaxomicin; treating the solution with nanofiltration concentrate, and separating to obtain fidaxomicin crude product; conducting preparative column chromatography on the fidaxomicin crude product, eluting with an acid aqueous solution containing an organic solvent, and separating to obtain the refined fidaxomicin product.
BIOSYNTHESIS OF EVERNINOMICIN ANALOGS IN MICROMONOSPORA CARBONACEA VAR AURANTIACA
Disclosed are methods of preparing everninomicin analogs by genetic alteration of Micromonospora carbonacea. Everninomicin analogs prepared by these methods and methods of using these analogs to treat infections are also disclosed.
BIOSYNTHESIS OF EVERNINOMICIN ANALOGS IN MICROMONOSPORA CARBONACEA VAR AURANTIACA
Disclosed are methods of preparing everninomicin analogs by genetic alteration of Micromonospora carbonacea. Everninomicin analogs prepared by these methods and methods of using these analogs to treat infections are also disclosed.
BIOSYNTHETIC GENE CLUSTER OF CARRIMYCIN
The present disclosure provides a biosynthetic gene cluster of carrimycin. The biosynthetic gene cluster comprises 44 gene open reading frames (orf), i.e., 5 orfs (orf10-14) encoding polyketide synthase, 9 orfs (orf1, 4-6, 15 and 36-39) related to polyketone synthesis extension unit and modification, 16 orfs (orf9, 16-22, 24, 26, 28, 29, 33-35 and 41) related to glycosyl synthesis, 6 orfs (orf7, 8, 30-32 and 40) related to glycosyl transfer, 2 orfs (orf3 and 25) related to resistance, 4 orfs (orf2, 23, 27 and 42) possibly related to regulation, a tsr resistance marker gene orf (orf43) and a 4-mycaroseglucoside isovaleryl transferase gene orf (orf44).
BIOSYNTHETIC GENE CLUSTER OF CARRIMYCIN
The present disclosure provides a biosynthetic gene cluster of carrimycin. The biosynthetic gene cluster comprises 44 gene open reading frames (orf), i.e., 5 orfs (orf10-14) encoding polyketide synthase, 9 orfs (orf1, 4-6, 15 and 36-39) related to polyketone synthesis extension unit and modification, 16 orfs (orf9, 16-22, 24, 26, 28, 29, 33-35 and 41) related to glycosyl synthesis, 6 orfs (orf7, 8, 30-32 and 40) related to glycosyl transfer, 2 orfs (orf3 and 25) related to resistance, 4 orfs (orf2, 23, 27 and 42) possibly related to regulation, a tsr resistance marker gene orf (orf43) and a 4-mycaroseglucoside isovaleryl transferase gene orf (orf44).
METHODS AND COMPOSITIONS FOR RECOMBINASE-BASED GENETIC DIVERSIFICATION
Compositions and methods using shufflon recombinases are presented for use in generating genetic diversity in molecules of interest.
CYTOCHROME P450 ENZYME
The invention provides an enzyme with cytochrome P450 activity with mutations at the 109 and/or 198 positions based on P450.sub.SeuC10 from Streptomyces eurythermus NRRL 2539. The invention extends to related nucleic acid material and microorganisms, kits containing the enzyme, methods of producing the enzyme and methods of using the enzyme to generate oxidised organic compounds.
CYTOCHROME P450 ENZYME
The invention provides an enzyme with cytochrome P450 activity with mutations at the 109 and/or 198 positions based on P450.sub.SeuC10 from Streptomyces eurythermus NRRL 2539. The invention extends to related nucleic acid material and microorganisms, kits containing the enzyme, methods of producing the enzyme and methods of using the enzyme to generate oxidised organic compounds.
Process for the preparation of fidaxomicin
The present invention provides an improved process for the preparation of fidaxomicin by culturing Actinoplanes deccanensis in the culture medium of the invention wherein the fidaxomicin is produced in a yield of greater than 500 mg/L broth. The present invention also provides a whole broth extraction process for the isolation of fidaxomicin from the fermentation broth. The present invention also provides fidaxomicin having purity of greater than 97% area by HPLC.