Abstract: The invention provides human lyases (HLYA) and polynucleotides which identify and encode HLYA. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with aberrant expression of HLYA.

Abstract: The invention relates to an immunogenic composition, characterized in that it comprises an adenyl cyclase-hemolysin (AC-Hly) protein, or an immunogenic portion of this AC-Hly, of a strain of Bordetella chosen from B. pertussis, B. parapertussis or B bronchiseptica, and in that it comprises, in addition, a bacterial extract containing the expression products of the vrg genes of a strain of Bordetella chosen from B. pertussis, B. parapertussis or B bronchiseptica, or a portion of these expression products which is sufficient to induce an immune response in a host to which the extract might be administered.

Abstract: The present inventors have discovered that Mg-chelatase is essential for the growth of Arabidopsis. Specifically, the inhibition of Mg-chelatase CHL H gene expression in Arabidopsis seedlings results in varying levels of chlorosis (yellowing), significantly reduced growth and developmental abnormalities. Thus, Arabidopsis Mg-chelatase can be used as a target for the identification of herbicides. Accordingly, the present invention provides methods for the identification of compounds that modulate Arabidopsis Mg-chelatase expression or activity, comprising: contacting a compound with a Arabidopsis Mg-chelatase, or a subunit thereof, and detecting the presence and/or absence of binding between said compound and said Mg-chelatase, or detecting a change in Mg-chelatase expression or activity. The methods of the invention are useful for the identification of herbicides and other compounds that can modulate plant growth and development.

Abstract: Process is provided for preparing a non-proteinogenic L-amino acid by means of an enzymic biotransformation in which O-acetyl-L-serine is reacted with a nucleophilic compound, while using an O-Acetyl-L-serine sulfhydrylase as catalyst, to give a non-proteinogenic L-amino acid. The process is carried out at a pH in the range between pH 5.0 and 7.4.

Abstract: Disclosed herein are structure-based modelling methods for the preparation of acetohydroxy acid synthase (AHAS) variants, including those that exhibit selectively increased resistance to herbicides such as imidazoline herbicides and AHAS inhibiting herbicides. The invention encompasses isolated DNAs encoding such variants, vectors that include the DNAs, and methods for producing the variant polypeptides and herbicide resistant plants containing specific AHAS gene mutations. Methods for weed control in crops are also provided.

Abstract: The present invention relates to citrate lyase polypeptides and to nucleic acid molecules coding for such polypeptides. Citrate lyase is transcriptionally regulated in adipose tissue, suggesting involvement in a novel pathway in energy expenditure, downstream of fatty acid metabolism. This makes CitE an attractive drug target for treating and/or preventing diseases such as obesity. Accordingly, the invention refers to the novel mammalian CitE-polypeptide, as well as homologs and functionally equivalent variants thereof. Further, the invention refers to the nucleic acid sequence encoding the novel protein and to vectors for expressing the protein in various organisms. The invention also refers to methods for expressing the protein in said organisms and for purifying the protein upon overexpression. Moreover, the invention refers to a methods wherein CitE is used for screening for substances that affect the activity of CitE.

Abstract: Compositions, methods and kits for diagnosing and treating cancer are provided. Therapeutic compositions may comprise agents that modulate the expression or activity of a sphingosine-1-phosphate lyase (SPL). Such compositions may be administered to a mammal afflicted with cancer. Diagnostic methods and kits may employ an agent suitable for detecting alterations in endogenous SPL. Such methods and kits may be used to detect the presence of a cancer or to evaluate the prognosis of a known disease. SPL polypeptides, polynucleotides and antibodies are also provided.

Abstract: Novel synthases and the corresponding nucleic acids encoding such synthases are disclosed herein. Such synthases possess an active site pocket that includes key amino acid residues that are modified to generate desired terpenoid reaction intermediates and products. Synthase modifications are designed based on, e.g., the three-dimensional coordinates of tobacco 5-epi-aristolochene synthase, with or without a substrate bound in the active site.

Abstract: The invention provides isolated nucleic acid molecules, designated DHY nucleic acid molecules, which encode novel DHY-related dehydratase molecules. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing DHY nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a DHY gene has been introduced or disrupted. The invention still further provides isolated DHY proteins, fusion proteins, antigenic peptides and anti-DHY antibodies. Diagnostic methods utilizing compositions of the invention are also provided.

Abstract: A method for disrupting epithelial barrier function in a host in need of the topical administration of a physiologically active substance which comprises applying to the epithelium of the host, barrier-disrupting amount of at least one agent selected from the group consisting of an inhibitor of ceramide synthesis, inhibitor of acylceramide synthesis, inhibitor of glucosylceramide synthesis, and inhibitor of sphingomyelin synthesis, an inhibitor of fatty acid synthesis, an inhibitor of cholesterol synthesis, a degradation enzyme of ceramides, acylceramide, glucosylceramides, sphingomyelin, an inhibitor of phospholipid, glycosphingolipid, including glucosylceramide, acylceramide or sphingomyelin degradation, and both inhibitors and stimulators of metabolic enzymes of free fatty acids, ceramide, and cholesterol, as well as a topical composition useful therefor are disclosed.

Abstract: The invention described herein relates to coenzymes useful for the synthesis of L-carnitine, particularly a compound of coenzyme A, and more particularly gamma-butyrobetainyl-coenzyme A and crotonobetainyl-coenzyme A, to procedures for their preparation and to their use for the production of L(−)-carnitine from crotonobetain and D(−)-carnitine.

Abstract: This invention relates to a process for the preparation of a 3-hydroxycarboxylic acid from a 3-hydroxynitrile. More specifically, 3-hydroxyvaleronitrile is converted to 3-hydroxyvaleric acid in high yield at up to 100% conversion, using as an enzyme catalyst 1) nitrile hydratase activity and amidase activity or 2) nitrilase activity of a microbial cell. 3-Hydroxyvaleric acid is used as a substitute for &egr;-caprolactone in the preparation of highly branched copolyester.

Abstract: The invention provides human lyases and associated proteins (HLYAP) and polynucleotides which identify and encode HLYAP. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with expression of HLYAP.

Abstract: The present invention relates to transformed bacterial hosts capable of expressing a pectate lyase enzyme endogenous to a strain of Thermotoga maritima, especially a Bacillus or E. coli host cell, is useful in a method for producing the Thermotoga maritima pectate lyase. The Thermotoga maritima pectate lyase is useful for industrial use, e.g. for treatment of textiles.

Abstract: Assays, preferably high throughput assays, for determining cysteine concentration, cysteine synthase activity, and identifying herbicides, fungicides, bactericides, and insecticides. Cysteine concentration is quantitated by contacting cysteine with a coumarin dye capable of conjugating with cysteine but not to O-acetyl serine or sulfide; exciting the conjugate with UV light; and detecting fluorescent light emitted by the conjugate. Cysteine synthase activity is determined by combining O-acetyl-L-serine, sulfide and cysteine synthase to form a reaction mixture under conditions suitable for cysteine production; contacting the reaction mixture with an appropriate coumarin dye; subjecting the reaction mixture to UV light; and detecting fluorescent light emission.

Abstract: Novel synthases and the corresponding nucleic acids encoding such synthases are disclosed herein. Such synthases possess an active site pocket that includes key amino acid residues that are modified to generate desired terpenoid reaction intermediates and products. Synthase modifications are designed based on, e.g., the three-dimensional coordinates of tobacco 5-epi-aristolochene synthase, with or without a substrate bound in the active site.

Abstract: The invention provides isolated animal soluble adenylyl cyclase and methods of modulating its expression and activity. Also provided are methods of utilizing soluble adenylyl cyclase for diagnosing pathological conditions and monitoring blood gases.

Abstract: A method of preparing 1,5-D-anhydrofructose in large quantities includes treating &agr;-1,4-glucan with a substantially pure &agr;-1,4-glucan lyase, which has been isolated from algae alone, wherein 1,5-D-anhydrofructose is produced directly from the &agr;-1,4-glucan.

Abstract: Compositions, methods and kits for diagnosing and treating cancer are provided Therapeutic compositions may comprise agents that modulate the expression or activity of a sphingosine-1-phosphate lyase (SPL). Such compositions may be administered to a mammal afflicted with cancer. Diagnostic methods and kits may employ an agent suitable for detecting alterations in endogenous SPL. Such methods and kits may be used to detect the presence of a cancer or to evaluate the prognosis of a known disease. SPL polypeptides, polynucleotides and antibodies are also provided.

Abstract: SPHINGLY polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilising SPHINGLY polypeptides and polynucleotides in therapy, and diagnostic assays for such.

Abstract: The present invention provides several methods for biological production of para-hydroxycinnamic acid (PHCA). The invention is also directed to the discovery of new fungi and bacteria that possess the ability to convert cinnamate to PHCA. The invention relates to developing of a new biocatalyst for conversion of glucose to PHCA by incorporation of the wild type PAL from the yeast Rhodotorula glutinis into E. coli underlining the ability of the wildtype PAL to convert tyrosine to PHCA. The invention is also directed to developing a new biocatalyst for conversion of glucose to PHCA by incorporation of the wildtype PAL from the yeast Rhodotorula glutinis plus the plant cytochrome P-450 and the cytochrome P-450 reductase into E. coli. In yet another embodiment, the present invention provides for the developing of a new biocatalyst through mutagenesis of the wild type yeast PAL which possesses enhanced tyrosine ammonia-lyase (TAL) activity.

Abstract: The present invention relates to a polypeptide comprising the amino acid sequence of SEQ ID NO:1 or an amino acid sequence having one or more amino acids deleted, replaced or added. The polypeptide further comprises polyester synthase activity. The invention further relates to a polyester synthase gene comprising DNA coding for a polypeptide, a recombinant vector comprising the gene, and a transformant transformed with the recombinant vector.

Abstract: A variant of a cell-wall degrading enzyme having a beta-helix structure, which variant holds at least one substituent in a position determined by identifying all residues potentially belonging to a stack; characterizing the stack as interior or exterior; characterizing the stack as polar, hydrophobic or aromatic/heteroaromatic based on the dominating characteristics of the parent or wild-type enzyme stack residues and/or its orientation relative to the beta-helix (interior or exterior); optimizing all stack positions of a stack either to hydrophobic aliphatic amino acids, hydrophobic aromatic or polar amino acids by allowing mutations within one or all positions to amino acids belonging to one of these groups; measuring thermostability of the variants by DSC or an application-related assay such as a Pad-Steam application test; and selecting the stabilized variants. Variant of a wild-type parent pectate lyase (EC 4.2.2.

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 picromycin. The combinatorial library is useful as a source of pharmaceutically active compounds. In addition, novel polyketides and antibiotics are prepared using this method.

Abstract: An assay to screen potential inhibitors of the C4 acid cycle in plants. The assay involves testing inhibition of C4 enzymes by incubating a mixture that includes pyruvate orthophosphate dikinase, phosphoenolpyruvate carboxylase and malate dehydrogenase and their substrates with a potential inhibitor. Detection of inhibition depends on comparing the resulting levels of NADH or NAD+ in the test mixture to the levels of NADH or NAD+ in a control.

Abstract: The invention relates to a nucleic acid molecule, comprising a nucleic acid which codes for a polypeptide with chorismate mutase activity and derivatives thereof, whereby the derivatives have at least 10% of the chorismate mutase activity of the chorismate mutase, according to the identification number of the SEQ ID NO:2. The invention further relates to vectors containing nucleic acid molecules, to host cells comprising nucleic acid molecules and their use in methods for producing polypetides with chorismate mutase activity. The invention also relates to the polypeptides with chorismate mutase activity and antibodies which specifically recognize the same. In addition, the invention relates to methods for producing auxotrophic yeast strains using the nucleic acid molecules and to the preparation of the yeast strains.

Abstract: Novel synthases and the corresponding nucleic acids encoding such synthases are disclosed herein. Such synthases possess an active site pocket that includes key amino acid residues that are modified to generate desired terpenoid reaction intermediates and products. Synthase modifications are designed based on, e.g., the three-dimensional coordinates of tobacco 5-epi-aristolochene synthase with or without a substrate bound in the active site.

Abstract: Compositions, methods and kits for diagnosing and treating cancer are provided. Therapeutic compositions may comprise agents that modulate the expression or activity of a sphingosine-1-phosphate lyase (SPL). Such compositions may be administered to a mammal afflicted with cancer. Diagnostic methods and kits may employ an agent suitable for detecting alterations in endogenous SPL. Such methods and kits may be used to detect the presence of a cancer or to evaluate the prognosis of a known disease. SPL polypeptides, polynucleotides and antibodies are also provided.

Abstract: The invention relates to the stabilization of enzymes through the formation of matrix stabilized enzyme crystals. A matrix stabilized enzyme crystal is formed through cross-linking a polymer having one or more reactive moieties with an enzyme crystal using a low concentration of a multi-functional cross-linking reagent. The invention includes matrix stabilized enzyme crystals of phenylalanine ammonia lyase and a method of using to treat hyperphenylalaninemia.

Abstract: A gene which contains the nucleotide sequence shown in SEQ ID NO:1 from nucleotide 671 to nucleotide 6295 or a nucleotide sequence which can be obtained therefrom by substitution, insertion or deletion of up to 30%, preferably up to 10%, particularly preferably up to 20%, especially preferably up to 5%, of the nucleotides, and whose gene product has the enzymatic activity of an adenylate cyclase, and its use.

Abstract: A method is disclosed for inhibiting carboxyltransferase with bisubstrate analogs. One such analog has been shown to inhibit the carboxyltransferase component of E. coli acetyl-CoA carboxylase. It is also expected to inhibit mammalian acetyl-CoA carboxylase, and thereby to act as an antiobesity agent and an anti-cancer agent.

Abstract: In order to optimize the flux or flow of carbon intermediates from normal cellular metabolism into PHAs it is desirable to optimize the expression of the enzymes of the PHA biosynthetic pathway. Gene fusions are genetic constructs where two open reading frames have been fused into one and encode hybrid proteins and in some cases bifunctional hybrid enzymes. Linkers may be added to spatially separate the two domains of the hybrid protein. In the case of enzymes which catalyse successive reactions in a pathway, the fusion of two genes results in bringing two enzymatic activities into close proximity to each other. When the product of the first reaction is a substrate for the second one, this new configuration of active sites may result in a faster transfer of the product of the first reaction to the second active site with a potential for increasing the flux through the pathway.

Abstract: This invention related to an isolated nucleic acid fragment encoding a tryptophan biosynthetic enzyme. This invention also related to the construction of a Chimeric gene encoding all or a portion of the tryptophan biosynthetic enzyme, in sense or antisense orientation, wherein expression of the Chimeric gene results in the production of altered levels of the tryptophan biosynthetic enzyme in a transformed host cell. The tryptophan biosynthetic enzymes include anthranilate phosphoribosyltransferase (E.C. 2.4.2.18) from corn (Zea mays), soybean (Glycine max) and wheat (Triticum aestivum); indole-3-glycerol phosphate synthase (E.C. 4.1.1.48) from corn (Zea mays), rice (Oryza sativa), soybean (Glycine max), and wheat (Triticum aestivum); and phosphoribosylanthranilate isomerase (E.C. 5.3.1.24) from corn (Zea mays), rice (Oryza sativa), (and wheat (Triticum aestivum).

Abstract: This invention provides novel genes encoding enzymes which decompose difficult-to-decompose thiophene compounds. By using these genes, sulfur atoms can be released from the thiophene compounds in fossil fuel such as petroleum, and the diffusion of sulfur into the environment caused by the combustion of the fossil fuel can be prevented.

Abstract: A process for production of L-methionine &ggr;-lyase crystals by using polyethylene glycol, characterized by comprising the first step of warming a solution containing L-methionine &ggr;-lyase before or after addition of polyethylene glycol thereto and the second step of adding an inorganic salt.

Abstract: In order to provide technical information necessary for the production of L-phenylalanine ammonia-lyase (PAL) by utilizing genetic engineering techniques, the structural gene for PAL and the amino acid sequence of PAL have been elucidated in Rhodosporidium toruloides, and novel recombinant DNA plasmids (e.g., pSW101, pYtrp6 and pKY201) have been created by inserting a DNA strand coding for the PAL gene between the 3′-terminus of the promoter region and the 5′-terminus of the terminator region. Moreover, transformants having such a novel recombinant DNA plasmid have been created, and a process for the production of PAL by growing such a novel transformant so as to cause PAL to be produced and accumulated in the culture has been established. Furthermore, there has been established a novel technique for the production of L-phenylalanine by reacting an ammonia donor with cinnamic acid in the presence of the PAL prepared by the aforesaid novel process.

Abstract: A bioengineered synthesis scheme for the production of shikimic acid from a carbon source is provided. Methods of producing shikimic acid from a carbon source based on the synthesis scheme are also provided.

Abstract: Novel polypeptides and the corresponding nucleic acids encoding such polypeptides are disclosed herein. The invention provides methods of making modified polypeptides by altering one or more amino acid residues involved in the active site of a preselected polypeptide.

Abstract: The present invention provides a purified and isolated nucleic acid encoding mycobacterial isocitrate lyase, as well as mutated forms of the nucleic acid. Further provided are purified and isolated isocitrate lyase proteins and mutated isocitrate lyase proteins. Additionally, the present invention provides vectors which comprises nucleic acid sequences encoding mycobacterial isocitrate lyase and mutated forms of this nucleic acid, as well as host cells containing these vectors. Also provided is a mycobacterium containing one or more mutations in its isocitrate lyase gene. Further provided by the present invention are agents that inhibit the activity or expression of a mycobacterial lyase protein, a method of identifying these, and a method of producing them. Finally, the present invention also provides a method of identifying genes required for persistence of mycobacteria.

Abstract: A DNA sequence encoding a human adenylyl cyclase is described. The amino acid sequence of the adenylyl cyclase is also described.

Abstract: The present inventors have discovered that chorismate mutase and chorismate synthase are essential for plant growth. Specifically, the inhibition of chorismate mutase or chorismate synthase gene expression in plant seedlings results in severe chlorosis, reduced growth and developmental abnormalities. Thus, in one aspect the invention provides compositions for the modulation of plant growth or development comprising chorismate synthase and chorismate mutase antisense and sense polynucleotides, dsRNA and ribozymes, and related expression cassettes and vectors. The compositions of the invention are particularly useful for the modulation and inhibition of plant growth. The invention further provides plants, plant cells, and seeds containing the polynucleotides of the invention. The inventors have proven that chorismate synthase and chorismate mutase can be used as targets for the identification of herbicides.

Abstract: The present invention provides dihydropicolinate synthase and dihydrodipicolinate reductase enzymes from Bacillus methanolicus, polynucleotides encoding the enzymes, and methods of producing L-lysinse in microorganisms expressing the enzymes.

Abstract: The construction of multicopy expression systems for methioninase are disclosed. The higher expression systems employ a promoter operably linked to two or more copies of a tandemly repeated methioninase encoding sequence. Such multicopy expression systems were found to produce unexpectedly high levels of methioninase when expressed in an appropriate host.

Abstract: Compositions and methods for enhancing disease resistance to a pathogen in a plant are provided. Methods of the invention comprise stably transforming a plant with an antisense nucleotide sequence for a gene encoding an enzyme in the C-5 porphyrin metabolic pathway and operably linking said antisense sequence to a pathogen-inducible promoter, such that invasion of a cell by a pathogen elicits a hypersensitive-like response that results in confinement of the pathogen to cells of initial contact. Transformed plants and seeds are provided. Nucleotide sequences encoding a wild-type maize urod gene useful in the present invention and the amino acid sequence for the protein encoded thereby are provided. These compositions are also useful for regulating cell death in specifically targeted tissues. A maize lesion mimic, dominant mutant phenotype, designated Les22, and the molecular basis for its manifestation are also provided.

Abstract: This invention relates to an isolated nucleic acid fragment encoding a branched-chain biosynthetic enzyme. The invention also relates to the construction of a chimeric gene encoding all or a portion of the branched-chain biosynthetic enzyme, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the branched-chain biosynthetic enzyme in a transformed host cell.

Abstract: cDNAs encoding, E-&agr;-bisabolene synthase, &dgr;-selinene synthase and &ggr;-humulene synthase from Grand Fir (Abies grandis) have been isolated and sequenced, and the corresponding amino acid sequences have been determined. Accordingly, isolated DNA sequences (SEQ ID No:12; SEQ ID No:19 and SEQ ID No:23) are provided which code for the expression of E-&agr;-bisabolene synthase (SEQ ID No:13), &dgr;-selinene synthase (SEQ ID No:20) and &ggr;-humulene synthase (SEQ ID No:24), respectively, from Grand Fir (Abies grandis). In other aspects, replicable recombinant cloning vehicles are provided which code for E-&agr;-bisabolene synthase, &dgr;-selinene synthase and &ggr;-humulene synthase, or for a base sequence sufficiently complementary to at least a portion of E-&agr;-bisabolene synthase, &dgr;-selinene synthase or &ggr;-humulene synthase DNA or RNA to enable hybridization therewith.

Abstract: The enzyme 2,3-dihydroxybenzoic acid decarboxylase has industrial applications for the decarboxylation of ring mounted carboxyls, specifically the decarboxylation of 2,3-dihydroxybenzoic acid to catechol. This invention relates to the isolation of a nucleic acid sequence from Aspergillus niger that encodes an enzyme that decarboxylates 2,3-dihydroxybenzoic acid. This invention further discloses the nucleic acid sequence, the protein sequence, vectors comprising the nucleic acid sequence, cells transformed with the nucleic acid sequence, and methods for the production of 2,3-dihydroxybenzoic acid decarboxylase.

Abstract: The present invention relates to the isolation of two DNA promoters from a coffee plant. The isolated promoters, one inducible and one constitutive, are capable of inducing the expression of a second DNA operably linked to the promoter. The present invention also relates to host cells, expression systems and transgenic plants containing the promoters of the invention.

Abstract: The present invention provides amino acid sequences of peptides that are encoded by genes within the human genome, the enzyme peptides of the present invention. The present invention specifically provides isolated peptide and nucleic acid molecules, methods of identifying orthologs and paralogs of the enzyme peptides, and methods of identifying modulators of the enzyme peptides.

Abstract: Pectin degrading enzymes derived from or endogeneous to Bacillus licheniformis or other Bacillus species which are at least 99% homologous to Bacillus licheniformis based on aligned 16S rDNA sequences have optimum activity at pH higher than 8. The pectin degrading enzymes belongs to the enzyme classes pectate lyases (EC 4.2.2.2), pectin lyases (EC 4.2.2.10) and polygalacturonases (EC 3.2.1.15) and are useful in industrial processes under alkaline conditions such as in textile processing and as an active ingredient eg in laundry detergents and hard surface cleaning products.