Abstract: A process is provided for the bioconversion of glycerol to 1,3-propanediol in which genes from a bacteria known to possess a diol dehydratase enzyme for 1,2-propanediol degradation are cloned into a bacterial host and the host is grown in the presence of glycerol; expression of the foreign genes in the host cell facilitates the enzymatic conversion of glycerol to 1,3-propanediol which is isolated from the culture.

Abstract: A process for the preparation of five-membered or six-membered ring lactams from aliphatic .alpha.,.omega.-dinitriles has been developed. In the process an aliphatic .alpha.,.omega.-dinitrile is first converted to an ammonium salt of an .omega.-nitrile-carboxylic acid in aqueous solution using a catalyst having an aliphatic nitrilase (EC 3.5.5.7) activity, or a combination of nitrile hydratase (EC 4.2.1.84) and amidase (EC 3.5.1.4) activities. The ammonium salt of the .omega.-nitrilecarboxylic acid is then converted directly to the corresponding lactam by hydrogenation in aqueous solution, without isolation of the intermediate .omega.-nitrilecarboxylic acid or .omega.-aminocarboxylic acid. When the aliphatic .alpha.,.omega.-dinitrile is also unsymmetrically substituted at the .alpha.-carbon atom, the nitrilase produces the .omega.-nitrilecarboxylic acid ammonium salt resulting from hydrolysis of the .omega.

Abstract: The present invention provides a nitrile hydratase nucleic acid fragment isolated from Pseudomonas putida which encodes a nitrile hydratase activity capable of catalyzing the hydrolysis of certain racemic nitriles to the corresponding R- or S-amides. Also provided are transformed microorganisms capable of the active expression of said nitrile hydratase activity. Additionally, the invention provides a transformant harboring the nitrile hydratase gene in conjunction with an amidase gene, both of which may be co-expressed producing active nitrile hydratase and amidase enzymes respectively. Methods for the production of such enantiomeric materials are also provided.

Abstract: The present invention provides the amino acid sequence and base sequence of a Pseudonocardia thermophila-derived nitrile hydratase, provides further a method for changing its amino acid sequence and base sequence without substantially changing the functions of said nitrile hydratase, and nitrile hydratases having a base sequence and an amino acid sequence as changed on the basis of said method, and provides furthermore a recombinant plasmid having the gene of said nitrile hydratase, a transformant containing said recombinant plasmid, a method of using said transformant for producing said enzyme, and a method of using said transformant for producing the corresponding amide compound from a nitrile compound.

Abstract: There are described methods for the synthesis of quinoid organic compounds from a renewable energy source such as glucose. The method comprises enhancing the amount of glucose equivalents introduced into the pathway, blocking the common pathway so as to accumulate dehydroquinate and converting the dehydroquinate to quinic acid.

Abstract: The present invention is a method for controlling biopolymer synthesis by determining the genetics and enzymology of polyhydroxybutyrate (PHB) biosynthesis at the molecular level. The purified enzymes and genes provide the means for developing new PHB-like biopolymers having polyester backbones. Specific aims are to 1) control the chain length of the polymers produced in fermentation processes through genetic manipulation, 2) incorporate different monomers into the polymers to produce copolymers with different physical properties, and 3) examine the physical/rheological properties of these new biopolymers in order to develop further design criteria at the molecular level.

Abstract: The present invention relates to a gene derived from Pseudomonas chlororaphis B23 strain which encodes a polypeptide having nitrile hydratase activity being capable of hydrating nitriles to amides. The invention also relates to a recombinant DNA containing the gene, and a transformant transformed with the recombinant DNA. The present invention further relates to a method of producing nitrile hydratase using the transformant and of amides using nitrile hydratase.

Abstract: The present invention provides a human ATP synthase subunit (ASYS) and polynucleotides which encode ASYS. The invention also provides expression vectors, host cells, agonists, antisense molecules, antibodies, or antagonists. The invention also provides methods for producing ASYS and for treating disorders associated with expression of ASYS.

Abstract: A crystallizable, purified chondroitinase ABC having a molecular weight of about 100,000 dalton by the measurement of the SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and the measurement by the gel permeation chromatography method, having alanine as the N-terminal amino acid and proline as the C-terminal amino acid is disclosed. A process for the purification of the crystallizable purified chondroitinase ABC comprising removing nucleic acid from an surfactant solution extract obtained from cells of chondroitinase ABC-producing microorganisms and chromatographically treating by concentration gradient elution using a weak cation exchange resin or a strong cation exchange resin is disclosed. A composition comprising a chondroitinase and serum albumin, gelatin, or a nonionic surfactant is disclosed. The enzyme is isolated from Proteus vulgaris ATCC 6896.

Abstract: A method for detecting autoantibodies to glutamic acid decarboxylase (GAD) in the serum of a patient as diagnostic of a diabetic or prediabetic condition in the patient, comprises contacting a serum sample from the patient with a GAD antigen and detecting binding of autoantibodies to GAD in the sample by the GAD antigen, wherein the GAD antigen comprises a GAD preparation containing an enhanced amount of dimer(s) or oligomer(s) of the 65 kD or 67 kD isoforms, or both, of GAD. A diagnostic kit is also inclosed.

Abstract: A crystallizable, purified chondroitinase ABC having a molecular weight of about 100,000 dalton by the measurement of the SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and the measurement by the gel permeation chromatography method, having alanine as the N-terminal amino acid and proline as the C-terminal amino acid is disclosed. A process for the purification of the crystallizable purified chondroitinase ABC comprising removing nucleic acid from an surfactant solution extract obtained from cells of chondroitinase ABC-producing microorganisms and chromatographically treating by concentration gradient elution using a weak cation exchange resin or a strong cation exchange resin is disclosed. A composition comprising a chondroitinase and serum albumin, gelatin, or a nonionic surfactant is disclosed. The enzyme is isolated from Proteus vulgaris ATCC 6896.

Abstract: The present invention provides the nucleotide sequences of Acetobacter operons, cdg operons encoding genes for the biosynthesis and degradation of cyclic diguanosine monophosphate (c-di-GMP). Specifically, the nucleotide sequences and deduced amino acid sequences of 3 phosphodiesterases isozymes, 3 diguanylate cyclase isozymes, and 2 polypeptides of unidentified function are provided. Also provided for are various strains of microorganisms, including Acetobacter cells genetically manipulated so as to produce elevated and/or reduced levels of one or more cdg operon encoded proteins.

Abstract: 2-Ketoaldonic acid is synthesized by aldolase condensation reaction involving pyruvate and an aldose acceptor in the presence of excess pyruvate. After the reaction is substantially complete, the excess pyruvate is removed from the reaction mixture by treatment with pyruvate decarboxylase.

Abstract: S-phenyl-L-cysteine can be produced in a high yield by reacting thiophenol and L-serine under the action of tryptophan synthase at a pH in a range of from 9.0 to 10.5. Purification of S-phenyl-L-cysteine obtained by this enzyme reaction can be effectively achieved by adjusting the pH of a crystal-containing reaction mixture to a strongly acidic level of 1.5 or lower to dissolve crystals of S-phenyl-L-cysteine, adding activated carbon to the pH-adjusted mixture, maintaining the resultant mixture at a temperature of from 20.degree. to 60.degree. C. under aeration with an oxygen-containing gas, subjecting the thus-obtained mixture to filtration, raising the pH of the resulting filtrate back to a range of from 2.5 to 6.0 to precipitate crystals of S-phenyl-L-cysteine and then recovering the thus-precipitated crystals.

Abstract: Recombinant methods and materials useful in producing lactoperoxidases are disclosed. An illustrative form of lactoperoxidase is the bovine protein shown in FIG. 1. FIG. 1 also shows the DNA sequence natively encoding the bovine lactoperoxidase, including contiguous regions of the gene. Such DNAs are useful in a variety of applications including antisense technology, formation of triple helices, and performance of diagnostic assays.

Abstract: This invention relates to stabilization of phenylalanine ammonia lyase against proteolytic degradation by chemical modification with crosslinking agents, or by genetic modification, a phenylalanine ammonia lyase variant, a method of preparing the variant and a pharmaceutical composition containing phenylalanine ammonia lyase.

Abstract: Flavorant compounds and compositions for food substances are prepared in a food-acceptable manner by bio-conversion of a cysteine S-complex with a food-acceptable micro-organism, particularly a yeast or filamentous fungus, which, upon incubation with the complex, splits the complex at a terminal carboxyl group in a beta position to yield a product including a thiol and at least one metabolite compound, and the product is isolated.

Abstract: The present invention relates to a method of separating a protein, in particular an enzyme, from an aqueous solution of proteins, comprising (a) providing an aqueous mixture of proteins with a salt concentration at or below 1.5 Molar, to which a water soluble polymer has been added, and (b) recovery of the protein on crystalline form.

Abstract: The invention provides a novel GDP-mannose 4,6-dehydratase.

Abstract: The cloning of the heparinase gene from Flavobacterium Heparinum using the polymerase chain reaction is described. The Open Reading Frame (ORF) corresponded to 1152 base pairs encoding a precursor protein of MW 43,800 daltons. The amino acid sequence reveals a 20-residue leader peptide. The gene was expressed in two expression systems in E. coli.

Abstract: The present invention provides novel plant DNA sequences coding for native adenylosuccinate lyase (ADSL). Methods for using the complete or partial ADSL coding sequence as a probe for diagnostic, mapping and other purposes are taught. Generation of transformed host cells capable of expressing ADSL is also taught. Methods of using the transformed host cells are taught, including methods for recombinant production of ADSL enzymes. A method for using the plant ADSL enzyme to screen for inhibitors of ADSL activity is also provided.

Abstract: A method of producing an optically active amino acid is disclosed which comprises converting a mixture of an amino group-containing compound, e.g., an alkanediamine, and fumaric acid into an optically active amino acid by the action of a microorganism. The method is useful in industrially producing an optically active amino acid from the inexpensive starting materials, i.e., fumaric acid and an amino compound, under mild conditions of ordinary temperature and ordinary pressure.

Abstract: A method for regulating enzyme activity, which entails contacting one or more enzymes with a gas containing one or more noble gases or mixtures thereof.

Abstract: The invention is directed to isolated polypeptides useful in ameliorating autoimmune disease.

Abstract: The invention relates to an enzyme, exo-.alpha.-1,4-glucan lyase, capable of successively cleaving the terminal .alpha.-1,4-D-glucosidic bonds from the non-reducing ends of an .alpha.-1,4-glucan. The enzyme is selected from the group consisting of .alpha.-1,4-glucan lyase isolated from an alga and functional derivatives and analogs derived from a nucleotide sequence related to an alga .alpha.-1,4-glucan lyase. Further, a method of enzymatically cleaving the terminal .alpha.-1,4-D-glucosidic bonds from the non-reducing ends of an .alpha.-1,4-glucan, and the degradation product 1,5-anhydrofructose for use as an oxygen radical scavenger or anti-oxidant and/or as a sugar substitute, are disclosed. Additionally, a method of producing an enzyme capable of successively cleaving the terminal .alpha.-1,4-glucosidic bonds from the non-reducing ends of an .alpha.

Abstract: The present invention describes methods for using methioninase as an antitumor reagent in anti-methionine chemotherapy. Specifically, methioninase is used to slow or stop cell division which can be enhanced with competitive inhibitors of methionine. In addition, methioninase can be used in combination with chemotherapeutic agents to increase the therapeutic effectiveness of the agent by inducing cell cycle synchronization.

Abstract: The isolation and characterization of genes involved in proteolytic processing in species of the genus Pichia is described. The availability of such genes has enabled the generation of strains of Pichia which are deficient in proteolytic activity, which strains are useful as hosts for the expression of proteolytically sensitive recombinant products. The isolation and characterization of additional genes from species of the genus Pichia is also described, as well as uses therefore.

Abstract: The invention concerns a process for the production of dihydroxyacetone phosphate (DHAP) by enzymatic oxidation of glycerophosphate in the presence of glycerophosphate oxidase and an H.sub.2 O.sub.2 -decomposing enzyme such as catalase and also concerns the conversion of DHAP formed in situ in a coupled enzymatic aldol addition to produce carbohydrates or corresponding derivatives.

Abstract: The present invention describes the isolation and sequence of genes from Flavobacterium heparinum encoding heparin and heparan sulfate degrading enzymes, heparinase II and heparinase III (EC 4.2.2.8). It further describes a method of expressing and an expression for heparinases I, II and III using a modified ribosome binding region derived from a promoter from glycosaminoglycan lyase genes of F. heparinum. Also, a multi-step protein purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography and hydroxylapatite chromatography is outlined. Antibodies against a post-translational modification moiety common to Flavobacterium heparinum proteins and a method to obtain antibodies specific to these moieties and to the amino acid sequences of heparinases I, II and III are described.

Abstract: Isolated polypeptides useful in ameliorating GAD-associated autoimmune disease as well as diagnostic and therapeutic methods of using the peptides are disclosed.

Abstract: A preparation of recombinant 3-hydroxy-3-methylglutaryl-CoA synthase is disclosed, wherein the preparation has at least 0.024 unitsg specific activity. Preferably, the preparation has at least 0.24 units/mg specific activity and is a crude cell extract. Preferably, at least 90% of the synthase molecules have not been substantially proteolytically cleaved. A preparation of recombinant HMG-CoA synthase wherein the preparation retains 50% activity after storage at 4.degree. C. for six months is also disclosed. A method of evaluating the efficacy of candidate anti-isoprene and anti-cholesterol drugs is also disclosed which comprises exposing the candidate drug to a recombinant 3-hydroxy-3-methylglutaryl CoA synthase preparation.

Abstract: A method for preparing N-acetylneuraminic acid characterized in reacting N-acetylglucosamine with pyruvic acid in an alkaline condition by the action of N-acetylneuraminic acid lyase.

Abstract: A method for controlling and modifying biopolymer synthesis by manipulation of the genetics and enzymology of synthesis of polyhydroxybutyrate (PHB) and polyhydroxyalkanoate (PHA) polyesters at the molecular level in procaryotic and eukaryotic cells, especially plants. Examples demonstrate the isolation, characterization, and expression of the genes involved in the production of PHB and PHA polymers. Genes encoding the enzymes in the PHB and PHA synthetic pathway (beta-ketothiolase, acetoacetyl-CoA reductase and PHB polymerase or PHA polymerase) from Zoogloea ramigera strain I-16-M, Alcaligenes eutrophus, Nocardia salmonicolur, and Psuedomonas olevarans were identified or isolated and expressed in a non-PHB producing organism, E. coli. Specific modifications to the polymers include variation in the chain length of the polymers and incorporation of different monomers into the polymers to produce co-polymers with different physical properties.

Abstract: The present invention relates to a gene derived from Pseudomonas chlororaphis B23 strain which encodes a polypeptide having nitrile hydratase activity being capable of hydrating nitriles to amides. The invention also relates to a recombinant DNA containing the gene, and a transformant transformed with the recombinant DNA. The present invention further relates to a method of producing nitrile hydratase using the transformant and of amides using nitrile hydratase.

Abstract: The nucleic acid coding for an .alpha.-acetolactate synthase from Lactococcus is provided, as well as vectors containing this nucleic acid and the use of these vectors for transforming microorganisms in which the production of .alpha.-acetolactate will be promoted, The nucleic acid comprises one or the other or both of a first segment corresponding to the ilvB gene (which encodes one subunit of .alpha.-acetolactate synthase of Lactococcus lactis subsp. lactis) and a second segment corresponding to the ilvN gene (which encodes a second subunit of .alpha.-acetolactate synthase of Lactococcus lactis subsp. lactis).

Abstract: The invention relates to modified materials based on polyacrylonitrile having amidic groups on their surface. The modification gives the material greater hydrophilic characteristics improving its comfort properties. In addition, it permits the polyacrylonitrile to be dyed also with acidic dyes thus making it possible for it to be used for the preparation of yarns mixed with natural fibres, such as wool for example. The process for their production involves treatment of the material with enzymes of the nitrile hydratasis class obtained from Brevibacterium imperiale.

Abstract: A method for preparing an active and inactive adenylate cyclase from a culture of Bordetella parapertussis having specific reactivity with polyclonal or monoclonal antibodies to adenylate cyclase from Bordetella pertussis is presented. The inactive adenylate cyclase is devoid of calmodulin-activatable adenylate cyclase activity and of affinity for calmodulin. The method comprises culturing a clone of Bordetella parapertussis, homogenizing the culture to produce a homogenate, and isolating the active and inactive adenylate cyclase from the homogenate by urea precipitation. The isolated active and inactive adenylate cyclase may be used to prepare a vaccine for the prevention of Bordetella infection.

Abstract: Processes are provided for removal of cells as cell pellets from liquid milk samples, or from cultures or extracts of other food materials or other materials of biological origin. The concentrated cells in the pellet can be analyzed by various techniques to determine the relative cell count, such as by lysing of the cells followed by measurement of ATP. Nucleic amplification, as by the polymerase chain reaction method, can be carried out using the cellular pellet directly, without need for isolation of nucleic acid from the cells. After the amplification, an assay can be carried out for amplified nucleic acid segment indicative of the presence of cells of interest in the sample. The invention thus provides methods for obtaining cellular components from samples of milk, and cultures or extracts of other materials, including food materials, and for determining relative contamination of milk and such other materials by microorganisms. The invention also provides kits for carrying out its various methods.

Abstract: Aureobacterium barkerei strain KDO-37-2 (ATCC 49977) and KDO aldolase (EC 4.1.2.23) isolated therefrom are disclosed. The KDO aldolase is further disclosed to have a broad substrate specificity with respect to its reverse reaction, i.e. the condensation of aldoses with pyruvate to form a wide range of 2-keto-3-deoxy-onic acids, including 2-keto-3-deoxy-nonulosonic acid, 2-keto-3-deoxy-octulosonic acid, 2-keto-3-deoxy-heptulosonic acid, and 2-keto-3-deoxy-hexulosonic acid. In particular, 3-deoxy-D-manno-2-octulosonic acid (D-KDO), a vital component of lipopolysaccharides found in the bacterial outer membrane may be synthesized from D-arabinose and pyruvate in 67% yield.

Abstract: The invention relates to an enzyme capable of fragmenting a high molecular mass N-acetylheparosan. This enzyme was obtained with the strain Escherichia coli (K5), strain SEBR 3282.

Abstract: A DNA sequence encoding a novel effector enzyme referred to as a cardiac adenylyl cyclase is described. The amino acid sequence of the cardiac adenylyl cyclase encoded by that DNA sequence is also described.

Abstract: This invention relates to methods for the isolation and purification of the recombinantly expressed major protein component of chondroitinase ABC, which is referred to as "chondroitinase I", from Proteus vulgaris (P. vulgaris). This invention further relates to methods for the isolation and purification of the recombinantly expressed second protein component of chondroitinase ABC, which is referred to as "chondroitinase II", from P. vulgaris. These methods provide significantly higher yields and purity than those obtained by adapting for the recombinant enzymes the method previously used for isolating and purifying native chondroitinase I enzyme from P. vulgaris.

Abstract: An improved method for chemotherapy of mammalian malignant cells which have an absolute requirement for methionine but lack methylthioadenosine phosphorylase (MTAse). The method comprises detection of MTAse negative cells in a mammal, administration of methionine .gamma.-lyase in sufficient amounts to reduce the volume of MTAse negative cells in the mammal, and co-administration of methylthioadenosine in amounts sufficient to ensure the continued availability of methionine to the mammal's non-malignant cells. Means for detection of MTAse negative cells are provided. Means for production and use of recombinant chemotherapeutic agents are also provided.

Abstract: A single, reproducible scheme to simultaneously purify all three of the heparin lyases from F. heparinum to apparent homogeneity is disclosed herein. The kinetic properties of the heparin lyases have been determined as well as the conditions to optimize their activity and stability. Monoclonal antibodies to the three heparinases are also described and are useful for detection, isolation and characterization of the heparinases.

Abstract: The present invention provides compounds that function as hydrolytic enzyme inhibitors (inactivators) and substrates. These compounds are useful in assays to detect and measure levels of hydrolytic enzyme activity and are more particularly useful in treatment regimens for various disease states and conditions implicating the underlying specific hydrolytic enzyme. Examples of hydrolytic enzymes include, but are not limited to, phospholipases, lipases, esterases, proteases, etc.

Abstract: Pharmaceutical compositions for delivering an effective dose to a desired site of a heparinase. These compositions are based on the discovery that heparinase alone can inhibit angiogenesis. The effective dosage is dependent not only on the heparinase, but also on the method and means of delivery, which can be localized or systemic. For example, in some applications, as in the treatment of psoriasis or diabetic retinopathy, the inhibitor is delivered in a topical ophthalmic carrier. In other applications, as in the treatment of solid tumors, the inhibitor is delivered by means of a biodegradable, polymeric implant.

Abstract: The present invention relates, in general, to an enzyme that degrades oxalic acid. In particular, the invention relates to the enzyme oxalate decarboxylase and to a DNA sequence encoding same. The invention further relates to a recombinant molecule comprising the oxalate decarboxylase encoding sequence and to a host cell transformed therewith. In addition, the invention relates to a method of protecting a plant from the deleterious effects of oxalic acid and to a method of reducing the oxalic acid content of a plant.

Abstract: A method of catalyzing in vitro reactions using seeds containing enhanced amounts of enzymes is disclosed. The method involves adding transgenic, non-wild type seeds, preferably in a ground form, to a reaction mixture and allowing the enzymes in the seeds to increase the rate of reaction. By directly adding the seeds to the reaction mixture the method provides a solution to the expensive and problematic process of extracting and purifying the enzyme. Methods of treatment are also provided whereby a subject lacking a sufficient supply of an enzyme is administered the enzyme in the form of seeds containing enhanced amounts of the enzyme.

Abstract: A method for controlling and modifying biopolymer synthesis by manipulation of the genetics and enzymology of synthesis of polyhydroxybutyrate (PHB) and polyhydroxyalkanoate (PHA) polyesters at the molecular level in procaryotic and eukaryotic cells, especially plants. Examples demonstrate the isolation, characterization, and expression of the genes involved in the production of PHB and PHA polymers. Genes encoding the enzymes in the PHB and PHA synthetic pathway (beta-ketothiolase, acetoacetyl-CoA reductase and PHB polymerass or PHA polymerase) from Zoogloea ramigera strain I-16-M, Alcaligenes eutrophus, Nocardia salmonicolur, and P. olevorans were identified or isolated and expressed in a non-PHB producing organism, E. coli. Specific modifications to the polymers include variation in the chain length of the polymers and incorporation of different monomers into the polymers to produce co-polymers with different physical properties.

Abstract: A chromatographic process for the copurification of chondroitinase proteins useful in ocular surgery for non-surgical disruption of chondroitin sulfate, the molecule which mediates the attachment between the retina and vitreous body in the human eye. The process involves the use of ion exchange resins in conjunction with an affinity elution with chondroitin sulfate to afford copurification of the proteins.