Abstract: The invention provides a nucleic acid molecule comprising: (a) a nucleotide sequence as shown in SEQ ID No. 35; or (b) a nucleotide sequence which is the complement of SEQ ID No. 35; or (c) a nucleotide sequence which is degenerate with SEQ ID No. 35; or (d) a nucleotide sequence hybridising under conditions of high stringency to SEQ ID No. 35, to the complement of SEQ ID No. 35, or to a hybridisation probe derived from SEQ ID No. 35 or the complement thereof; or (e) a nucleotide sequence having at least 80% sequence identity with SEQ ID No. 35; or (f) a nucleotide sequence having at least 65% sequence identity with SEQ ID No. 35 wherein said sequence preferably encodes or is complementary to a sequence encoding a nystatin PKS enzyme or a part thereof.

Abstract: Provided are a process for producing purified anthocyanidin glucoside in which a rhamnose end of anthocyanidin rutinoside is cleaved using rhamnosidase to convert the anthocyanidin rutinoside component into anthocyanidin glucoside, the anthocyanidin glucoside component being then purified and isolated; or a crystalline anthocyanidin glucoside salt obtained by further crystallizing the purified anthocyanidin glucoside and a process for producing the same.

Abstract: Host cells, such as E. coli, are provided with an expression system for making starter units required for biosynthesis of polyketides using the ato pathway.

Abstract: The present invention provides an isolated polypeptide comprising a nucleotide sequence encoding a protein which is involved in midecamycin biosynthesis, wherein said protein comprises an amino acid sequence selected from SEQ ID NOs: 2 to 10, 13, 14, 16, 19, 20, 22 to 26, and 28 to 38 or a modified amino acid sequence of said amino acid sequence having one or more amino acid modifications without affecting activity of the protein.

Abstract: Described are host cells that contain a polyketide synthase gene and a thioesterase II gene, where the polyketide synthase gene has been modified to prevent utilization of its native starter unit for its expressed polyketide synthase. Also described are host cells containing a polyketide synthase gene and an endogenous thioesterase II gene, where the activity of the endogenous thioesterase II gene product has been decreased or eliminated. Methods for culturing these cells to produce polyketides are also provided, as are the polyketides produced.

Abstract: A method for enhancing a host cell's biosynthetic production of 2-KLG is described. Such method comprises selecting a host cell that has an at least partially intracellular synthetic pathway which utilizes 2,5-DKG to produce 2-KLG; increasing the transport of said 2,5-DKG into said host cell while maintaining the integrity of the host cell; culturing the host cell to produce said 2,5-DKG; and producing 2-KLG. The transport of the 2,5-DKG is increased by transforming into the host cell DNA encoding for one or more enzymes transporting the 2,5-DKG into the host cell.

Abstract: Biosyntheses of compounds whereof at least portions are polyketides produced by means of polyketide synthase (PKS) enzyme complexes are carried out after specific alterations have been made within the acyltransferase (AT) domains of the PKSs. Particular motifs in or near the substrate binding pocket are disclosed, such that alterations therein affect substrate specificity.

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 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: The present invention relates to hybrid glycosylated products, and in particular, to natural products such as polyketides and glycopeptides, and to processes for their preparation. The invention is particularly concerned with recombinant cells in which a cloned microbial glycosyltransferase can be conveniently screened for its ability to generate specific glycosylated derivatives when supplied with polyketide, peptide, or polyketide-peptides as substrates. The invention demonstrates that cloned glycosyltransferases when rapidly screened for their ability to attach a range of activated sugars to a range of exogenously supplied or endogenously generated aglycone templates, show a surprising flexibility towards both aglycone and sugar substrates, and that this process allows the production of glycosylated polyketides in good yield.

Abstract: The present invention provides a method capable of simultaneous processing of plural test samples for the receptor binding property of a chemical substance, which does not require immobilization of the receptor or a special device, and a reagent to be used for this method. That is a method for assaying the receptor binding property of an assay target substance is provided, the method comprising the steps of (a) competitively reacting a known concentration of a ligand and the assay target substance with a known concentration of the receptor in a solution, (b) measuring, without physically removing the ligand bound with the receptor prior to the assay, the amount of a free ligand in the solution using one or more antibodies against the ligand, and (c) determining the receptor binding property of the assay target substance using the amount of the free ligand as an index.

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: A novel pathway for the synthesis of polyhydroxyalkanoates is provided. A method of synthesizing a recombinant polyhydroxyalkanoate monomer synthase is also provided. These recombinant polyhydroxyalkanoate synthases are derived from multifunctional fatty acid synthases or polyketide synthases and generate hydroxyacyl acids capable of polymerization by a polyhydroxyalkanoate synthase. Also provided is a biosynthetic gene cluster for methymycin and pikomycin as well as a biosynthetic gene cluster for desosamine.

Abstract: Disclosed are nucleic acid and amino acid sequences for acetolactate synthase, acetolactate synthase regulatory regions, &agr;-tubulin promoter, a promoter from a Thraustochytriales polyketide synthase (PKS) system, and fatty acid desaturase promoter, each from a Thraustochytriales microorganism. Also disclosed are recombinant vectors useful for transformation of Thraustochytriales microorganisms, as well as a method of transformation of Thraustochytriales microorganisms. The recombinant nucleic acid molecules of the present invention can be used for the expression of foreign nucleic acids in a Thraustochytriales microorganism as well as for the deletion, mutation, or inactivation of genes in Thraustochytriales microorganisms.

Abstract: Genes and proteins involved in the biosynthesis of macrolides by microorganisms, in particular the nucleic acids forming the biosynthetic locus for the 16-member macrolide rosaramicin from Micromonospora carbonacea. These nucleic acids can be used to make expression constructs and transformed host cells for the production of rosaramicin. The genes and proteins allow direct manipulation of macrolides and related chemical structures via chemical engineering of the proteins involved in the biosynthesis of rosaramicin.

Abstract: Disclosed is a process for producing a high &agr;G-AA content product, which comprises the steps of: contacting a solution as a material solution, containing &agr;G-AA, L-ascorbic acid, and a saccharide(s), with an anion exchange resin packed in a column to adsorb the &agr;G-AA and L-ascorbic acid on the anion exchanger; washing the anion exchanger with water to remove the saccharide(s) therefrom; feeding to the column an aqueous solution, as an eluent, of an acid and/or a salt with a concentration of less than 0.5 N to fractionate a fraction rich in 2-O-&agr;-D-glucopyranosyl-L-ascorbic acid and a fraction rich in L-ascorbic acid; and collecting the former fraction.

Abstract: A biosynthetic gene cluster for methymycin and pikromycin as well as a biosynthetic gene cluster for desosamine is provided.

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: The present invention relates to glycosyl-etoposide prodrugs, a process for the preparation thereof and the use thereof in combination with functionalized tumor-specific enzyme conjugates for treating cancers.

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: The present invention provides a new bioactive compound of formula (I), which has an antimicrobial activity against pathogenic microorganisms, and a process for production thereof. Also provided are a pharmaceutical composition comprising the compound and pharmaceutically acceptable carrier, a use of the WF002 as a medicament and use of the compound for manufacture of the medicament for treatment of infectious disease.

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 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: The invention relates to a compound of formula (l) 1

Abstract: The present invention relates to a continuous process for preparing optically pure (S)-3,4-dihydroxybutyric acid derivatives expressed by the following Formula 1 and more particularly, to the continuous process which enables preparing optically pure (S)-3,4-dihydroxybutyric acid derivatives economically in large quantities, by: (a) Preparing &agr;-(1,4) linked oligosaccharide having adequate sugar distribution by reacting amylopectin which is easily available from natural product with enzyme under a specific condition; and (b) Performing oxidatioin by running basic anion exchange resin with an oxidant to give (S)-3,4-dihydroxybutyric acid-anion exchange resin complex, dissociating the (S)-3,4-dihydroxybutyric acid from anion exchange resin complex and esterification sequentially under a specific condition.  wherein Represents linear or branched alkyl group with 1˜5 carbon atoms.

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: Synthesis of an amino-disaccharide, amino-oligosaccharide or a derivative thereof, characterized in that a monosaccharide, a disaccharide, an oligosaccharide, a glycoside or a derivative thereof, in the presence of a glycosidase as catalyst, is reacted with an amino-deoxy-saccharide or a glycoside or derivative thereof, and that the amino-saccharide is optionally isolated from the product mixture directly or after chemical/enzymatic modification.

Abstract: The present invention has for its object to provide a method for producing an L-allysine acetal which involves a fewer steps and is efficient. This invention relates to method for producing an L-allysine acetal which comprises; converting a D,L-allysine acetal of the following general formula (1) (wherein R1 and R2 are the same or different, and each of them represents an alkyl group having 1 to 8 carbon atoms, or they combinedly form a ring and represent an alkylene group having 2 to 8 carbon atoms) to a mixture of a 2-oxo-6,6-dialkoxyhexanoic acid of the following general formula (2) (wherein R1 and R2 are as defined above) and an L-allysine acetal of the following general formula (3) (wherein R1 and R2 are as defined above) by reacting in the presence of an enzyme capable of stereoselective oxidative deamination of D-amino acids and; isolating said L-allysine acetal after said converting.

Abstract: The etiological agent responsible for non-A, non-B hepatitis has been identified and termed hepatitis C virus (HCV). The HCV genome is a linear, positive-stranded RNA molecule with a length of approximately 9,400 nucleotides. With the exception of rather short untranslated regions at the termini, the genome consists of one large, uninterrupted, open reading frame encoding a polyprotein of approximately 3,000 amino acids. This polyprotein has been shown to be cleaved co-translationally into individual viral structural and non-structural (NS) regions. The structural protein region is further divided into capsid (Core) and envelope (E1 and E2) regions. The NS regions are divided into NS-1 to NS-5. The present invention is directed toward peptides derived from the HCV NS4 region that contain immunologically important epitopes. A series of biotinylated peptides were prepared by solid phase peptide synthesis and these peptides have proven useful for the detection of HCV-specific antisera.

Abstract: A method is provided for determining in a medium the presence or amount of an analyte which is capable of binding to a ligand partner to form a ligand complex, which method comprises: (a) contacting the medium with either: (i) a ligand partner conjugated with an enzyme being capable of catalysing a reaction, or one or more reactions in a cascade thereof, to produce hydrogen peroxide, or (ii) a ligand partner and either a competing analyte or an analog of said analyte, a competing analyte or analyte analog being capable of forming a ligand complex with the ligand partner and the competing analyte or analyte analog being conjugated with an enzyme capable of catalysing a reaction, or one or more reactions in a cascade thereof, to produce hydrogen peroxide, (b) optionally separating complexed and uncomplexed enzyme conjugates, (c) causing or allowing the reaction or cascade of reactions to occur to produce hydrogen peroxide by contacting the complexed or uncomplexed enzyme conjugate with a corresponding enzyme su

Abstract: Methods of capturing and labeling a species, include attracting magnetically attractable particles to a solid support by magnetic forces, which particles have an affinity for the species, contacting the particles on the support with a sample containing the species to capture the species onto the particles on the support, and binding the species captured on the particles directly or indirectly to a detectable label before and/or whilst the species is captured on the particles on the support. The label may be bound to the captured species via an immunological binding partner which binds selectively to the species and may be a fluorescent label, luminescent label, enzyme label, dye label, phosphorescent label, metal-chelating label, radio label, spin label, heavy metal label, nucleic acid or nucleic acid analog hybridization label, avid or avid-like label suitably bound to or incorporated in particles which also bear a binding agent such as an antibody causing the particles to bind to the captured species.

Abstract: The invention relates in part to methods and compositions for identifying the presence, measuring the amount, stabilizing, and facilitating recovery of troponin complexes or individual troponin isoforms in a sample.

Abstract: The present invention relates to a method for the specific labeling of a protein containing selenocyst(e)ine and/or cyst(e)ine groups, comprising the following steps:at least one incubation of a protein-containing sample with at least one modifying agent specific for selenocyst(e)ine and/or cyst(e)ine groups, followed byat least one further incubation of said protein-containing sample with at least one labeling agent specific for selenocyst(e)ine and/or cyst(e)ine groups;wherein at least one substance interacting with said protein is added prior to and/or during and/or after at least one of said incubations.

Abstract: The present invention provides methods for the simultaneous measurement of triiodothyronine (T.sub.3) and thyroxine (T.sub.4) in biological fluids such as serum by direct equilibrium dialysis and immunoassay. Specifically, the method comprises dialyzing the serum sample to equilibrium in a physiological buffer system so that the free T.sub.3 and the free T.sub.4 are separated from T.sub.3 and T.sub.4 bound to serum proteins. The method further comprises combining a measured quantity of the dialyzed serum sample having free T.sub.3 and free T.sub.4 with reagents comprising a measured quantity of T.sub.3 labelled with a detectable marker and a measured quantity of T.sub.4 labelled with a detectable marker; an anti-T.sub.3 antibody of sufficient specificity and in sufficient quantity to bind a measurable quantity of the free T.sub.3, and an anti-T.sub.4 antibody of sufficient specificity and in sufficient quantity to bind a measurable quantity of the free T.sub.4.

Abstract: The present invention relates to compounds that are bis-biotins. These compounds comprise two biotinyl radicals connected by a chain of atoms, usually at least 16 atoms in length. The bis-biotin is conjugated to a member of a specific binding pair ("sbp member") wherein the chain is not part of the sbp member. Also disclosed are compositions comprising a complex of avidin and a bis-biotin as described above. The compounds and compositions of the invention find use in an assay for an analyte wherein there is employed a reagent system comprising an avidin reagent and a biotin reagent. The improvement of the present invention comprises using as the biotin reagent a bis-biotin as described above. Also disclosed are kits comprising the present bis-biotins and methods of preparing a bis-biotinylated conjugate of a member of a specific binding pair ("sbp member") for use in a specific binding assay.

Abstract: The present invention relates to a method for biotinylating a desired subset of a larger population of proteins, wherein the subset of proteins is first selectively bound to a solid phase, thereby separating it from undesired proteins of the population, and then, still bound to solid phase, it is biotinylated. Unreacted biotin may then be washed from the solid phase, and the biotinylated protein may be uncoupled from the solid phase.

Abstract: A method of determining affinity and kinetic properties for the solution interaction between an analyte and a binding partner therefor, which method comprises: (a) mixing a solution of said analyte with a solution of said binding partner, contacting the resulting reaction solution with (i) immobilised binding partner, or analogue, for said analyte, and/or (ii) immobilised analyte, or analogue, and monitoring the binding of said analyte and/or binding partner in said reaction solution to the respective immobilised species to determine the variation with time of the concentration of free analyte and/or binding partner in said solution; and/or (b) contacting a solution of the reaction complex of said analyte and a binding partner therefor with (i) immobilised binding partner, or analogue, and/or (ii) immobilised analyte, or analogue, and monitoring the binding of said analyte and/or binding partner in said reaction complex solution to the respective immobilised species to determine the variation with time of the

Abstract: The invention relates to a method for carrying out heterogeneous immunoassays, in particular to a method for separating a coated solid phase from a liquid phase by means of precipitation and subsequent centrifugation, with a detectable activity remaining in the liquid phase.

Abstract: Disclosed is a high throughput compatible assay that is useful for the identification of specific antagonists of TRAF-receptor interactions. The modular flexibility of the assay makes it possible to introduce simple modifications in order to measure the interaction of any TNF receptor cytoplasmic domain (or TRAF-binding protein) with any of the six TRAF proteins, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5 and TRAF6.

Abstract: Methods and compositions are provided for forming complexes between dsDNA and oligomers of heterocycles, aliphatic amino acids, particularly omega-amino acids, and a polar end group. By appropriate choice of target sequences and composition of the oligomers, complexes are obtained with low dissociation constants. The formation of complexes can be used for identification of specific dsDNA sequences, for inhibiting gene transcription, and as a therapeutic for inhibiting proliferation of undesired cells or expression of undesired genes.

Abstract: A biosensor apparatus for detecting a binding event between a ligand and receptor. The apparatus includes an electrode substrate coated with a high-dielectric hydrocarbon-chain monolayer, and having ligands attached to the exposed monolayer surface. Binding of a receptor to the monolayer-bound ligand, and the resultant perturbation of the monolayer structure, causes ion-mediated electron flow across the monolayer. In one embodiment, the monolayers have a coil-coil heterodimer embedded therein, one subunit of which is attached to the substrate, and the second of which carries the ligand at the monolayer surface.

Abstract: A method of determining the concentration of a sample antigen in the presence of an interferant by(1) running two immunoassays on the sample: one assay where the interferant influences the binding of both the sample antigen and a labeled antigen and a second assay where the interferant influences the binding of the sample antigen but not the labeled antigen;(2) obtaining a plot of the possible sample antigen concentrations versus the possible interferant concentrations corresponding to the readout for the sample for each of the two immunoassays; and(3) determining the sample antigen concentration and the interferant concentration which correspond to the point that appears in both the immunoassays plots.

Abstract: An arrangement for the investigation of hydrophilic macromolecules (1) in an aqueous solution having a solid carrier surface (5) onto which a lipid film (24) is disposed, wherein the molecules (1) to be investigated are bound, by means of a molecular coupling system (20), to the lipid film (24) and thus are immobilized, is characterized in that the molecular coupling system (20) comprises at least two, preferably three molecular or nuclear components (21, 22, 23), which can be coupled to each other, of which a first component (21) is bound to the lipid film (24) and a second component (22) is bound to a hydrophilic macromolecule (1) to be investigated. In this way, it is rendered possible to immobilize and investigate any hydrophilic macromolecules, wherein the coating of the solid carrier surface should be able to be carried out in an as easy as possible and reversible manner.

Abstract: The present invention provides a method of detecting antigens, which comprises immobilizing an antigen to a solid support and contacting the solid support with a means for hybridizing a labeled dendrimer to the antibody, through an oligonucleotide complexed thereto. A directly oligonucleotide labeled primary antibody or an oligonucleotide labeled secondary antibody may be employed, and a conventionally labeled dendrimer can subsequently be hybridized to the oligonucleotide through one or more of the outer arms of the dendrimer. The present invention offers the advantage over conventional methods of antigen detection by providing multiple label molecules per antigen, thereby enhancing the observed signal associated with the label.

Abstract: A method for determining the complexed forms of immunologically determinable prostate specific antigen (cPSA) in a blood sample, e.g., by two-site immunometric assays, in which the blood sample is treated to render free PSA (fPSA) immunologically nondetectable. A particularly preferred immunometric assay method employs three anti-PSA antibodies: an antibody that binds to both cPSA and fPSA (anti-tPSA), a second anti-tPSA antibody which is characterized by the unique property that binding to fPSA is blocked by binding of fPSA-specific antibodies, and a third antibody which is a fPSA-specific antibody. Thus, binding of the fPSA-specific antibody to PSA in the sample allows only cPSA to be measured in the immunometric assay. Measurement of cPSA blood levels has been found to provide a method for aiding in the diagnosis and monitoring of prostate cancer that is highly sensitive and specific, and eliminates the need for a significant number of patients to undergo unnecessary prostate biopsy.

Abstract: The present invention provides internally calibrated competitive assays for use on a solid support. Additionally, the invention provides a method of using such assays.

Abstract: This invention relates to an improvement in a method for detecting labeled molecules and especially biotinylated molecules and particularly relates to a method for reducing background signal problems in such detection methods.

Abstract: An assay device for detection or determination of an analyte in a sample uses opposable components and is suitable for assay of human chorionic gonadotropin and other protein or glycoprotein hormones.

Abstract: A method of detecting an antibody in a sample using a labelling compound and comprising the steps of mixing a ligand antigen, antibody or hapten bound to biotin with the sample; an antibody directed against the antibody to be detected bound to paramagnetic particles; and a chemiluminescent acridinium compound bound to avidin or streptavidin to form a solid phase complex; separating the solid phase from the liquid phase; and analyzing the separated solid phase for the presence of chemiluminescent complex.

Abstract: This invention relates to glycosyltransferase, genes encoding glycosyltransferase, recombinant vectors having such a gene, host cells transformed with such a recombinant vector, and uses thereof. This invention makes it possible to glycosylate indolopyrrolocarbazole derivatives conveniently and economically.