Abstract: Genetic engineering of the erythromycin polyketide synthase genes to effect combinatorial alterations of catalytic activities in the biosynthetic pathway can be used to generate a library of macrolides impractical to produce by chemical methods. The library includes examples of analogs with one, two and three altered carbon centers of the polyketide products.

Abstract: The invention provides compositions, including pharmaceuticals, comprising glycosylated kinamycins. The compositions and methods of the invention can be used to treat infections, i.e., as antibiotics, and as anti-tumor agents. The compositions of the invention can also act as act as electrophilic azo-coupling agents in vitro or in vivo. The invention also provides enzymes capable of generating kinamycin, nucleic acids that encode them, antibodies that bind to them, and methods for making and using them.

Abstract: Three distinct primary regions of acyltransferase (AT) domains of modular polyketide synthases (PKS) can be changed by site-specific mutagenesis of the corresponding coding sequences to change the specificity of the domain and the structure of the polyketide produced by the PKS.

Abstract: Disclosed are multibinding compounds which include macrolide antibiotics, aminoglycosides, lincosamides, oxazolidinones, streptogramins, tetracycline and/or other compounds at which bind to bacterial ribosomal RNA and/or to one or more proteins involved in ribosomal protein synthesis in the bacterium, which are useful in treating bacterial infections. The compounds adversely affect protein expression and have an antibacterial effect. The multibinding compounds of this invention containing from 2 to 10 ligands covalently attached to one or more linkers. Each ligand is macrolide antibiotic, aminoglycoside, lincosamide, oxazolidinone, streptogramin, tetracycline or other compound which binds to bacterial ribosomal RNA and/or one or more proteins involved in ribosomal protein synthesis in the bacterium.

Abstract: Combinatorial libraries of polyketides can be obtained by suitable manipulation of a host modular polyketide synthase gene cluster such as that which encodes the PKS for erythromycin. The combinatorial library is useful as a source of pharmaceutically active compounds. In addition, novel polyketides and antibiotics are prepared using this method.

Abstract: Streptomyces plasmids comprising two SCP2-derived replicons, at least one of which is derived from SCP2@, have a higher copy number and exhibit greater stability than SCP2- or SCP2*-derived plasmids having only a single SCP2-derived replicon and can be used to increase gene expression, relative to the latter plasmids, in host cells.

Abstract: Combinatorial libraries of polyketides can be obtained by suitable manipulation of a host modular polyketide synthase gene cluster such as that which encodes the PKS for erythromycin. The combinatorial library is useful as a source of pharmaceutically active compounds.

Abstract: The stereochemical centers of a polyketide can be changed by replacement of ketosynthase domains in the polyketide synthase (PKS) enzyme that produces the polyketide. The specificity of the AT domains of a PKS is determined by a hypervariable region that can be replaced or altered to change the specificity of the AT domain from a naturally occurring extender unit to another naturally or non-naturally occurring extender unit. Non-naturally occurring extender units, including methylmalonyl N-acetyl cysteamine thioester can be incorporated into polyketides in recombinant host cells or in cell-free systems to make polyketides.

Abstract: Disclosed is a family of P450 monooxygenases, each member of which regioselectively oxidizes avermectin to 4″-keto-avermectin. The P450 monooxgenases find use in methods and formulations for making emamectin from avermectin. Also disclosed are methods for purifying the P450 monooxygenases of the invention, binding agents that specifically bind to the P450 monooxygenases of the invention, and genetically engineered cells that express the P450 monooxygenases of the invention. Also disclosed are ferredoxins and ferredoxin reductases that are active with the P450 monooxygenases of the invention.

Abstract: Linking sequences which modulate cross-talk between modules of Type I polyketide synthases have been identified. Thus, arbitrarily chosen modules can be mixed and matched by supplying the appropriate linkers to obtain desired polyketide synthases and new polyketides. The modules are provided suitable linkers so that the polyketide chain is passed from one module to the other in the correct sequence. Synthetic peptides which mimic linkers can be used to inhibit the synthesis of polyketides. Kinetic channeling, both intrapolypeptide and interpolypeptide, of diketide intermediates in a Type I polyketide synthase can occur.

Abstract: The present invention provides combinatorial methods for rapidly generating a diverse library of glycorandomized structures, comprising incubating one or more aglycons and a pool of NDP-sugars in the presence of a glycosyltransferase. The glycosyltransferase may be one that is associated with or involved in production of natural secondary metabolites, or one which is putatively associated with or involved in production of natural secondary metabolites. The glycosyltransferase may show significant flexibility with respect to its NDP-sugar donors and/or its aglycons. NDP-sugar donors may be commercially available, or may be produced by utilizing mutant or wild type nucleotidyltransferases significant flexibility with respect to their substrates.

Abstract: Five protein families cooperate to form the warhead structure that characterizes enediyne compounds, both chromoprotein enediynes and non-chromoprotein enediynes The protein families include a polyketide synthase and thioesterase protein which form a polyketide synthase catalytic complex involved in warhead formation in enediynes. Genes encoding a member of each of the five protein families are found in all enediyne biosynthetic loci. The genes and proteins may be used in genetic engineering applications to design new enediyne compounds and in methods to identify new enediyne biosynthetic loci.

Abstract: Cell-free systems which effect the production of polyketides employing modular polyketide synthases are described. Libraries of new and/or known polyketides may also be produced in cell-free systems employing aromatic PKS, modular PKS or both.

Abstract: Modified PKS gene clusters which produce novel polyketides in an efficient system in a host cell or in a cell free extract are described.

Abstract: Combinatorial libraries of polyketides can be obtained by suitable manipulation of a host modular polyketide synthase gene cluster such as that which encodes the PKS for erythromycin. The combinatorial library is useful as a source of pharmaceutically active compounds. In addition, novel polyketides and antibiotics are prepared using this method.

Abstract: Recombinant DNA compounds that encode all or a portion of the oleandolide polyketide synthase are used to express recombinant polyketide synthase genes in host cells for the production of oleandolide, oleandolide derivatives, and polyketides that are useful as antibiotics and motilides.

Abstract: The present invention relates to polynucleotide molecules comprising nucleotide sequences encoding an aveC gene product, which polynucleotide molecules can be used to alter the ratio or amount of class 2:1 avermectins produced in fermentation cultures of S. avermitilis. The present invention further relates to vectors, host cells, and mutant strains of S. avermitilis in which the aveC gene has been inactivated, or mutated so as to change the ratio or amount of class 2:1 avermectins produced.

Abstract: The present invention relates to polynucleotide molecules comprising nucleotide sequences encoding the aveC gene product, which polynucleotide molecules can be used to alter the ratio or amount of class 2:1 avermectins produced in fermentation cultures of Streptomyces avermitilis. The present invention further relates to vectors, host cells, and mutant strains of Streptomyces avermitilis in which the aveC gene has been inactivated, or mutated so as to change the ratio or amount of class 2:1 avermectins produced.

Abstract: Host cells comprising recombinant vectors encoding the FK-520 polyketide synthase and FK-520 modification enzymes can be used to produce the FK-520 polyketide. Recombinant DNA constructs comprising one or more FK-520 polyketide synthase domains, modules, open reading frames, and variants thereof can be used to produce recombinant polyketide synthases and a variety of different polyketides with application as pharmaceutical and veterinary products.

Abstract: A multiple-plasmid system for heterologous expression of polyketides facilitates combinatorial biosynthesis. The method can be extended to any modular polyketide synthase (PKS) or non-ribosomal peptide synthase (NRPS) and has the potential to produce thousands of novel natural products, including ones derived from further modification of the PKS or NRPS products by tailoring enzymes.

Abstract: The use of enzymes which catalyze the production of starter and extender units for polyketides is described. In addition, modified loading modules are described, which can accept a variety of starting units such as substituted benzoates, and which can be used to generate substituted derivatives of natural products. These enzymes may be used to enhance the yield of polyketides that are natively produced or polyketides that are rationally designed. By using these techniques, the synthesis of a complete polyketide has been achieved in E. coli. Production can be enhanced in microbial organisms of polyketides and other secondary metabolites by delaying production until after exponential phase and by maintaining relatively constant nutrient levels in the medium. In addition, polyketide production can be enhanced by providing an expression system for a thioesterase II. Thus, by modifying the host organism and changing the culture conditions, synthesis of a secondary metabolite can be enhanced.

Abstract: The invention generally relates to polyunsaturated fatty acid (PUFA) polyketide synthase (PKS) systems isolated from or derived from non-bacterial organisms, to homologues thereof, to isolated nucleic acid molecules and recombinant nucleic acid molecules encoding biologically active domains of such a PUFA PKS system, to genetically modified organisms comprising PUFA PKS systems, to methods of making and using such systems for the production of bioactive molecules of interest, and to novel methods for identifying new bacterial and non-bacterial microorganisms having such a PUFA PKS system.

Abstract: The invention provides a biosynthetic gene cluster for mitomycin, for example, a mitomycin biosynthetic cluster from organisms such as Streptomyces, for instance, S. lavendulae, as well as methods of using gene(s) within the cluster to alter antibiotic biosynthesis and to prepare a polyketide synthase.

Abstract: Regions of sequence identity are recurrent in some modular polyketide synthase (PKS) gene clusters. Such sequences are potentially detrimental to the stability of PKS gene clusters and expression plasmids for the genes in the gene cluster. PKS gene and gene cluster stability can be improved, and reproducible polyketide titers can be obtained using those genes and gene clusters when the regions of sequence identity are reduced or eliminated by replacing one or more identical or homologous segments with non-homologous segments that encode the same or a substantially similar amino acid sequence.

Abstract: Recombinant host cells that comprise recombinant DNA expression vectors that drive expression of a product and a precursor for biosynthesis of that product can be used to produce useful products such as polyketides in host cells that do not naturally produce the product or produce the product or precursor at low levels due to the absence of the precursor or the presence of the precursor in rate limiting amounts.

Abstract: Hybrid and novel polyketide synthases and polyketides are produced by use of a multiple vector system. The combinatorial possibilities offered by placing the various catalytic activities of PKS systems on separate vectors permits the construction of improved libraries of PKS and polyketides. In addition, polyketides can be produced in hosts that ordinarily do not produce polyketides by supplying, along with an expression system for the desired PKS, an expression system for holo ACP synthase.

Abstract: Migrastatin and a migrastatin analog can be produced by fermentation of Streptomyces platensis NRRL 18993 and used in pharmaceutical formulations to treat cancer and/or inhibit metastasis of cancer cells.

Abstract: Novel polyketides and novel methods of efficiently producing both new and known polyketides, using recombinant technology, are disclosed. In particular, a novel host-vector system is described which is used to produce polyketide synthases which in turn catalyze the production of a variety of polyketides.

Abstract: The present invention recognizes that marine organisms comprise nucleic acid molecules that encode polypeptides that catalyze the synthesis of bioactive compounds, such as polyketides including bryopyran rings, such as bryostatins. One aspect of the present invention is a composition including at least one nucleic acid molecule that encodes at least one polypeptide that catalyzes at least one step in the synthesis of at least one polyketide such as a bryopyran ring, wherein said at least one nucleic acid molecule is derived from at least one marine organism. A second aspect of the present invention is a composition including a library of nucleic acid molecules of the present invention. These nucleic acid molecules can be used in a combinatorial biosynthesis of polyketides, bryopyran rings and bryostatins.

Abstract: Combinatorial libraries of polyketides can be obtained by suitable manipulation of a host modular polyketide synthase gene cluster such as that which encodes the PKS for erythromycin. The combinatorial library is useful as a source of pharmaceutically active compounds. In addition, novel polyketides and antibiotics are prepared using this method.

Abstract: Compounds useful in the inhibition of HIV integrase, the prevention or treatment of infection by HIV and the treatment of AIDS, either as compounds, pharmaceutically acceptable salts, pharmaceutical composition ingredients, whether or not in combination with other antivirals, immunomodulators, antibiotics or vaccines are described. Methods of treating AIDS and methods of preventing or treating infection by HIV are also described. The culture Actinoplanes sp. MA7220 (ATCC 202188) is also disclosed, as well as processes for making compounds of the present invention employing the culture.

Abstract: The compound referred to herein as IB-962 12, having structure (I) can be obtained by cultivating the strain of Micromonospora sp. ES25-008, available under the accession number CECT-3333, and can be hydrolyzed to give IB-96212B having structure (II). The sugar substituent which is L-rhodinose, can its elf be derivatized or the sugar can be replaced, in either case, giving further derivatives of IB-96212 having a group other than L-rhodinose at the position of the sugar.

Abstract: The present invention relates to compounds made by a subset of modules from one or more polyketide synthase (“PKS”) genes that are used as starting material in the chemical synthesis of novel molecules, particularly naturally occurring polyketides or derivatives thereof. The biologically derived intermediates (“bio-intermediates”) generally represent particularly difficult compounds to synthesize using traditional chemical approaches due to one or more stereocenters. In one aspect of the invention, an intermediate in the synthesis of epothilone is provided that feeds into the synthetic protocol of Danishefsky and co-workers. In another aspect of the invention, intermediates in the synthesis of discodermolide are provided that feed into the synthetic protocol of Smith and co-workers.

Abstract: The use of enzymes which catalyze the production of starter and extender units for polyketides is described. In addition, modified loading modules are described, which can accept a variety of starting units such as substituted benzoates, and which can be used to generate substituted derivatives of natural products. These enzymes may be used to enhance the yield of polyketides that are natively produced or polyketides that are rationally designed. By using these techniques, the synthesis of a complete polyketide has been achieved in E. col. This achievement permits a host organism with desirable characteristics to be used in the production of such polyketides and to assess the results of gene shuffling.

Abstract: Generic overproduction host cells can be used to produce any polyketide and obviate the need for performing conventional strain improvement.

Abstract: The stereochemical centers of a polyketide can be changed by replacement of ketosynthase domains in the polyketide synthase (PKS) enzyme that produces the polyketide. The specificity of the AT domains of a PKS is determined by a hypervariable region that can be replaced or altered to change the specificity of the AT domain from a naturally occurring extender unit to another naturally or non-naturally occurring extender unit. Non-naturally occurring extender units, including methylmalonyl N-acetyl cysteamine thioester can be incorporated into polyketides in recombinant host cells or in cell-free systems to make polyketides.

Abstract: Novel polyketides and novel methods of efficiently producing both new and known polyketides, using recombinant technology, are disclosed. In particular, a novel host-vector system is described which is used to produce polyketide synthases which in turn catalyze the production of a variety of polyketides.

Abstract: The heterologous expression of the OlePKS in Streptomyces lividans, produces 8,8a-deoxyoleandolide, an aglycone precursor of oleandomycin. The co-expression with DEBS in S. lividans of the P450 monooxygenase OleP produces 8,8a-dihydroxy-6-deoxyerythonolide B and other derivatives that are precursors to important macrolide antibiotics.

Abstract: Recombinant nucleic acids that encode all or a portion of the epothilone polyketide synthase (PKS) are used to express recombinant PKS genes in host cells for the production of epothilones, epothilone derivatives, and polyketides that are useful as cancer chemotherapeutics, fungicides, and immunosuppressants.

Abstract: The present invention relates to polynucleotide molecules comprising nucleotide sequences encoding an aveC gene product, which polynucleotide molecules can be used to alter the ratio or amount of class 2:1 avermectins produced in fermentation cultures of S. avermitilis. The present invention further relates to vectors, host cells, and mutant strains of S. avermitilis in which the aveC gene has been inactivated, or mutated so as to change the ratio or amount of class 2:1 avermectins produced.

Abstract: Spinosyn biosynthetic genes, spinosyn producing microorganisms transformed with the biosynthetic genes, methods using the biosynthetic genes to increase production of spinosyn insecticidal macrolides, and methods using the genes or fragments thereof to change the products produced by spinosyn-producing microorganisms.

Abstract: Hybrid and novel polyketide synthases (PKSs) and polyketides are produced by use of a multiple vector system. The combinatorial possibilities offered by placing the various catalytic activities of PKS systems on separate vectors permits the construction of improved libraries of PKS and polyketides. In addition, polyketides can be produced in hosts that ordinarily do not produce polyketides by supplying, along with an expression system for the desired PKS, an expression system for holo acyl carrier protein (ACP) synthase.

Abstract: Recombinant DNA compounds that encode all or a portion of the oleandolide polyketide synthase are used to express recombinant polyketide synthase genes in host cells for the production of oleandolide, oleandolide derivatives, and polyketides that are useful as antibiotics and motilides.

Abstract: The present invention relates to polynucleotide molecules comprising nucleotide sequences encoding an aveC gene product, which polynucleotide molecules can be used to alter the ratio or amount of class 2:1 avermectins produced in fermentation cultures of S. avermitilis. The present invention further relates to vectors, host cells, and mutant strains of S. avermitilis in which the aveC gene has been inactivated, or mutated so as to change the ratio or amount of class 2:1 avermectins produced.

Abstract: The present invention is directed to compositions and methods for producing avermectins, and is primarily in the field of animal health. The present invention relates to the identification and characterization of two novel genes, herein referred to as the aveR1 and aveR2 genes, that are involved in regulating avermectin polyketide synthase (PKS) expression and avermectin biosynthesis in Streptomyces avermitilis. The present invention is based on the discovery that inactivation of these genes results in an increase in the amount of avermectin produced by S. avermitilis.

Abstract: Yields of polyketides produced in host cells such as Streptomyces can be increased by coexpression of the ptpA gene. The introduction of recombinant vectors encoding ptpA and polyketide synthase genes is more efficient and does not require methyl-free DNA if the host cell is a restriction/methylation deficient strain, such as Streptomyces lividans K4-114, K4-155, or K27-39.

Abstract: An antigenically active hybrid glutamic acid decarboxylase (GAD) comprising an amino-terminal moiety derived from the GAD67 isoform linked directly or indirectly with a middle and carboxy-terminal moiety derived from the GAD65 isoform, and production thereof as a recombinant protein by expression in eukaryotic host cells, particularly yeasts.

Abstract: The invention provides methods and compositions relating to DNA Fragmentation Factor (DFF) polypeptides and related nucleic acids. More particularly, the present invention provides the sequence for the active subunit of DFF. The polylpeptides may be produced recombinantly from host cells transformed from the disclosed DFF encoding nucleic acids or purified from human cells. The invention provides isolated DFF hybridization probes and primers capable of specifically hybridization with the disclosed DFF genes, DFF-specific binding agents such as specific antibodies, and methods of making and using the subject compositions.

Abstract: Processes for the production of isoflavones are described wherein plant material from plants of the genus leguminosae are contacted with water, an enzyme which cleaves isoflavone glycosides to the aglucone form and a C.sub.2 -C.sub.10 organic solvent, so as to form a combination, incubating the combination for a time sufficient to allow isoflavones of the aglucone form to partition into the organic solvent component, and thereafter recovering isoflavones from the organic solvent component.

Abstract: Spinosyn biosynthetic genes, spinosyn producing microorganisms transformed with the biosynthetic genes, methods using the biosynthetic genes to increase production of spinosyn insecticidal macrolides, and methods using the genes or fragments thereof to change the products produced by spinosyn-producing microorganisms.