Abstract: The invention is directed to a novel purified mouse C5-epimerase, fragments thereof, nucleic acids encoding the same and the recombinant production thereof. The invention is also directed to fragments of such epimerase, especially N-terminal fragments that are useful in fusion protein constructs to enhance the activity of recombinantly-produced heterologous epimerase enzymes.

Abstract: Disclosed are 23 genes, termed “GEP” genes, found in Streptococcus pneumonia, which are located within operons that are essential for survival. Also disclosed is a related essential gene found in Bacillus subtilis. These genes and the polypeptides that they encode, as well as homologs thereof, can be used to identify antibacterial agents for treating bacterial infections such as streptococcal pneumonia.

Abstract: The invention relates to the use of the N-acetyl amino acid racemase from Amycolatopsis orientalis subspecies lurida for the racemiszation of N-carbamoyl amino acids. This use permits the 100% preparation of optically pure amino acids starting from racemic hydantoins in an enzymatic overall process.

Abstract: The invention relates to an isolated polynucleotide having a polynucleotide sequence which codes for the alr gene, and a host-vector system having a coryneform host bacterium in which the air gene is present in attenuated form and a vector which carries at least the air gene according to SEQ ID No 1, and the use of polynucleotides which comprise the sequences according to the invention as hybridization probes.

Abstract: The present invention relates to a 5-substituted hydantoin racemase, which efficiently catalyzes racemization reactions at a high optimum temperature for racemization reactions, DNA coding for the racemase, and processes for producing optically active amino acids.

Abstract: A method for racemizing with N-acylamino acid racemase (NAAR) derived from Sebekia benihana and a method for producing optically active amino acids using the racemaization method are provided. The racemase of the present invention can efficiently catalyze the racemization of acylamino acid substrates including N-acylalanine, N-acylaspartic acid, N-acylleucine, and N-acylvaline. Furthermore, this method can be applied to efficient production of optically active amino acids, which are useful, for example, as medicinal raw materials.

Abstract: The present invention relates to isolated DNA sequences which code for the expression of plant 1-deoxy-D-xylulose-5-phosphate reductoisomerase protein, such as the sequence presented in SEQ ID NO:1 which encodes a 1-deoxy-D-xylulose-5-phosphate reductoisomerase protein from peppermint (Mentha x piperita). Additionally, the present invention relates to isolated plant 1-deoxy-D-xylulose-5-phosphate reductoisomerase protein. In other aspects, the present invention is directed to replicable recombinant cloning vehicles comprising a nucleic acid sequence which codes for a plant 1-deoxy-D-xylulose-5-phosphate reductoisomerase, to modified host cells transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence of the invention.

Abstract: The present invention relates to the processes of racemization and deprotection of special N-protected amino acids in the acylase/racemase system for the total conversion of special N-protected racemic amino acids into optically pure amino acids.

Abstract: There are provided a DNA coding for phosphohexuloisomerase, which is a protein defined in the following (A) or (B), and a method for producing the enzyme:

Abstract: The invention relates to DNA which encodes Arabidopsis 1-eoxy-xylulose-5-phosphate reductoisomerase and to a method of identifying modulators of the 1-deoxy-D-xylulose-5-phosphate reductoisomerase and 1-deoxy-D-xylulose-5-phosphate synthase activity.

Abstract: The invention relates to a polypeptide having (1) a ligand domain that specifically binds to a molecule on an external surface of a cell, and (2) a DNA binding domain of a Topoisomerase I. The polypeptide optionally contains a translocation domain to facilitate DNA delivery into a cell.

Abstract: The present invention provides a process for producing biologically active recombinant yeast protein disulfide isomerase, which includes the steps of: a) deleting, substituting, or adding one or more bases in a region encoding an endoplasmic reticulum retention signal in a gene encoding protein disulfide isomerase of yeast to modify the gene so as not to encode part or all of the endoplasmic reticulum retention signal; b) incorporating the modified gene into an expression vector; c) transforming host cells with the expression vector; and d) culturing the host cells transformed with the expression vector in a culture medium kept at a nearly neutral pH, thereby causing protein disulfide isomerase to be secreted in an active state outside the host cells. According to the process of the invention, recombinant yeast protein disulfide isomerase can be secreted in a large amount in a biologically active state into a culture medium, and can be collected by a simple purification method.

Abstract: An isolated polynucleotide comprising a polynucleotide sequence chosen from the group consisting of a) polynucleotide which is identical to the extent of at least 70% to a polynucleotide which codes for a polypeptide which comprises the amino acid sequence of SEQ ID No. 2, b) polynucleotide which codes for a polypeptide which comprises an amino acid sequence which is identical to the extent of at least 70% to the amino acid sequence of SEQ ID No. 2, c) polynucleotide which is complementary to the polynucleotides of a) or b), and d) polynucleotide comprising at least 15 successive nucleotides of the polynucleotide sequence of a), b) or c), and a process for the fermentative preparation of L-amino acids using coryneform bacteria in which at least the gpmB gene is present in enhanced form, and the use of polynucleotides which comprise the sequences according to the invention as hybridization probes.

Abstract: Disclosed are isolated polypeptides having glucose isomerase activity selected from: (a) a polypeptide having an amino acid sequence which has at least 95% identity with amino acids of SEQ ID NO:2; (b) a variant of the polypeptide having an amino acid sequence of SEQ ID NO:2 comprising a substitution, deletion, and/or insertion of one or more amino acids; (c) a fragment of (a) that has glucose isomerase activity; and (d) a polypeptide having a pH optimum in the range of 5.7 to 6.3 at 60° C., a pH optimum in the range of 6.1 to 6.7 at 90° C and a temperature optimum of above 90° C. Also disclosed are isolated nucleic acid sequences encoding the polypeptides, nucleic acid constructs, vectors, and host cells comprising the nucleic acid sequences as well as methods for producing and using the polypeptides.

Abstract: A UDP-galactose epimerase enzyme is described. In addition, there is described a nucleotide sequence coding for the same.

Abstract: The present invention is directed to an N-acetyl amino acid racemase (AAR) as well as the gene coding for it, to a plasmid, vector and microroganism containing the gene. A method of producing enantiomer-enriched amino acids, and derivatives thereof, is also provided.

Abstract: Disclosed herein are expression systems that make use of Ero1 to enhance disulfide bond formation and thereby to increase the yield of properly folded recombinant proteins. Also disclosed herein are recombinant Ero1 polypeptides, nucleic acids, vectors, and cells for expressing such polypeptides.

Abstract: The present invention relates to protein disulfide isomerases which are encoded by a nucleic acid sequence which hybridizes with (i) the DNA sequence of SEQ ID NO:1 or (ii) the DNA sequence of SEQ ID NO:2, under the following conditions: presoaking in 5×SSC and prehybridizing for 1 h at ˜40° C. in a solution of 5×SSC, 5×Denhardt's solution, 50 mM sodium phosphate, pH 6.8, and 50 &mgr;g of denatured sonicated calf thymus DNA, followed by hybridization in the same solution supplemented with 50 &mgr;Ci 32-P-dCTP labelled probe for 18 h at ˜40° C. followed by washing three times in 2×SSC, 0.2% SDS at 40° C. for 30 minutes; and fragments thereof. The present invention also relates to DNA sequences encoding the protein disulfide isomerases, compositions comprising said protein disulfide isomerases and methods of use thereof.

Abstract: A biosynthetic method for producing vitamin C (ascorbic acid, L-ascorbic acid, or AA) is disclosed. Such a method includes fermentation of a genetically modified microorganism or plant to produce L-ascorbic acid. In particular, the present invention relates to the use of microorganisms and plants having at least one genetic modification to increase the action of an enzyme involved in the ascorbic acid biosynthetic pathway. Included is the use of nucleotide sequences encoding epimerases, including the endogenous GDP-D-mannose:GDP-L-galactose epimerase from the L-ascorbic acid pathway and homologues thereof for the purposes of improving the biosynthetic production of ascorbic acid. The present invention also relates to genetically modified microorganisms, such as strains of microalgae, bacteria and yeast useful for producing L-ascorbic acid, and to genetically modified plants, useful for producing consumable plant food products.

Abstract: Hexulose phosphate synthase was purified from Mycobacterium gastri to determine an amino acid sequence of the enzyme. Oligonucleotide primers to be used for PCR were synthesized on the basis of obtained amino acid sequence information. A DNA fragment, which was amplified by performing PCR by using a template of genomic DNA prepared from Mycobacterium gastri, was used as a probe so that colony hybridization was carried out with respect to a library comprising fragments obtained by digesting Mycobacterium gastri genomic DNA with PstI to obtain a positive clone. Thus, the DNA fragment which contains ORF encoding hexulose phosphate synthase was obtained. Furthermore, the DNA fragment cloned contained other ORF(ORF-1). The expression system of ORF-1 was constructed (pT-HPIS-1), and it was introduced into Escherichia coli cell. After the induction of the expression of ORF-1, the hexulose phosphate isomerase activity was found in the extract of cells harvoring pT-HPIS-1.

Abstract: A Vaccinia topoisomerase-adapted linker, which contains an oligonucleotide primer binding site of known sequence, useful for specifically joining the linker to the end of a polynucleotide of unknown sequence and allowing subsequent PCR amplification of the polynucleotide or DNA is provided. Kits containing the invention Vaccinia topoisomerase-adapted linker and one or more linker-specific oligonucleotides for annealing to the linker in PCR are also provided. In addition, the invention provides methods for using the invention linkers in linker-mediated PCR amplification procedures for isolation and optional sequencing of isolated PCR amplification products.

Abstract: The invention provides YfiI pseudouridine synthase polypeptides and polynucleotides encoding YfiI pseudouridine synthase polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing YfiI pseudouridine synthase polypeptides to screen for antibacterial compounds.

Abstract: The present invention relates to a novel ppGpp synthetase and expression systems for improved production of proteins of interest in Gram-positive bacteria, involving use of the novel ppGpp synthetase.

Abstract: An L-ribose isomerase which isomerizes aldoses such as L-ribose, D-lyxose, D-talose, D-mannose, L-allose and L-gulose into their corresponding ketoses such as L-ribulose, D-xylulose, D-tagatose, D-fructose, L-psicose and L-sorbose. The enzymatic reaction is a reversible equilibrium reaction. The L-ribose isomerase can be obtained from microorganisms of the genus Acinetobacter.

Abstract: The present invention relates to isolated nucleic acid sequences encoding polypeptides having cellobiose dehydrogenase activity. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the nucleic acid sequences as well as recombinant methods for producing the polypeptides.

Abstract: The present invention relates to isolated DNA sequences which code for the expression of plant 1-deoxy-D-xylulose-5-phosphate reductoisomerase protein, such as the sequence presented in SEQ ID NO:1 which encodes a 1-deoxy-D-xylulose-5-phosphate reductoisomerase protein from peppermint (Mentha x piperita). Additionally, the present invention relates to isolated plant 1-deoxy-D-xylulose-5-phosphate reductoisomerase protein. In other aspects, the present invention is directed to replicable recombinant cloning vehicles comprising a nucleic acid sequence which codes for a plant 1-deoxy-D-xylulose-5-phosphate reductoisomerase, to modified host cells transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence of the invention.

Abstract: The invention relates to inhibitors of PPIase activity and pharmaceutical compositions containing such inhibitors. Such compounds are useful for treatment of disorders characterized by inappropriate cell proliferation. In particular the compounds disclosed herein inhibit the activity of members of the Pinl/parvulin class of PPIases. These compounds have been designed based on the high resolution X-Ray derived crystal structure of the human enzyme Pin1.

Abstract: Topoisomerase III polypeptides and DNA and RNA encoding such Topoisomerase III polypeptides and a procedure for producing such polypeptides by recombinant techniques is disclosed. Also disclosed are methods for utilizing such Topoisomerase III for the treatment of infection, particularly bacterial infections. Antagonists against such Topoisomerase III and their use as a therapeutic to treat infections, particularly bacterial infections are also disclosed. Also disclosed are diagnostic assays for detecting diseases related to the presence of Topoisomerase III nucleic acid sequences and the polypeptides in a host. Also disclosed are diagnostic assays for detecting polynucleotides encoding Streptococcal Topoisomerase El and for detecting the polypeptide in a host.

Abstract: Topoisomerase I polypeptides and DNA and RNA encoding such Topoisomerase I polypeptides and a procedure for producing such polypeptides by recombinant techniques is disclosed. Also disclosed are methods for utilizing such Topoisomerase I for the treatment of infection, particularly bacterial infections. Antagonists against such Topoisomerase I and their use as a therapeutic to treat infections, particularly bacterial infections are also disclosed. Also disclosed are diagnostic assays for detecting diseases related to the presence of Topoisomerase I nucleic acid sequences and the polypeptides in a host. Also disclosed are diagnostic assays for detecting polynucleotides encoding Staphylococcal Topoisomerase I and for detecting the polypeptide in a host.

Abstract: Camptothecin compounds having a C10 11 O—(CF2)nO group where n is 1 or 2 are effective anti-tumor compounds. These compounds inhibit the enzyme topoisomerase I and may alkylate DNA of the associated topoisomerase I-DNA cleavable complex.

Abstract: Topoisomerase I polypeptides and DNA and RNA encoding such topoisomerase I polypeptides and a procedure for producing such polypeptides by recombinant techniques is disclosed. Also disclosed are methods for utilizing such topoisomerase I for the treatment of infection, particularly bacterial infections. Antagonists against such topoisomerase I and their use as a therapeutic to treat infections, particularly bacterial infections are also disclosed. Also disclosed are diagnostic assays for detecting diseases related to the presence of topoisomerase I nucleic acid sequences and the polypeptides in a host. Also disclosed are diagnostic assays for detecting polynucleotides encoding streptococcal topoisomerase I and for detecting the polypeptide in a host.

Abstract: Purified &bgr;-amino acids are of considerable interest in the preparation of pharmacologically active compounds. Although enantiomerically pure &bgr;-amino acids, such as L-&bgr;-lysine, can be produced by standard chemical synthesis, this traditional approach is time consuming, requires expensive starting materials, and results in a racemic mixture which must be purified further. However, DNA molecules encoding lysine 2,3-aminomutase can be used to prepare L-&bgr;-lysine by methods that avoid the pitfalls of chemical synthesis. In particular, L-&bgr;-lysine can be synthesized by cultures of host cells that express recombinant lysine 2,3-aminomutase. Alternatively, such recombinant host cells can provide a source for isolating quantities of lysine 2,3-aminomutase, which in turn, can be used to produce L-&bgr;-lysine in vitro.

Abstract: The invention provides isolated nucleic acid compounds encoding a glycosyltransferase enzyme of Amycolatopsis orientalis. Also provided are vectors carrying genes that encode said enzyme, transformed heterologous host cells for expressing said enzyme, and methods for producing glycopeptide compounds using the cloned genes that encode said enzyme.

Abstract: There is provided a gene for 2-aminothiazoline-4-carboxylate (ATC) racemase, a recombinant DNA comprising the gene or DNA fragment thereof, a transformant or transductant comprising the recombinant DNA, and a process for preparing the 2-aminothiazoline-4-carboxylate racemase using the transformant or transductant. According to the invention, the ATC racemase may be efficiently produced by microorganisms such as E. coli and Pseudomonas and L-cysteine and L-cystine may be efficiently synthesized using the enzyme.

Abstract: This invention provides novel assays for topoisomerase activity and for identifying topoisomerase inhibitors. The assays include both solid-phase and liquid-phase methods that are amenable to high throughput screening methods. The assays of the invention are readily adaptable to numerous types of topoisomerase, and are capable of identifying novel classes of topoisomerase activity modulators.

Abstract: The present invention relates to methods and compositions for the modulation of bacterial topoisomerase enzymes within bacterial cells. More specifically, the present invention relates to bacterial assays wherein the levels of bacterial topoisomerase enzymes or the levels of target sites within the enzymes are varied within bacterial test strains in order to screen for compounds that target, i.e., interact with the topoisomerase enzymes, causing DNA damage and hence, bacterial growth inhibition and/or cell death. The present methods and compositions are useful for the identification and characterization of novel therapeutic antibacterial compounds.

Abstract: The invention provides isolated nucleic acid compounds encoding the stem peptide biosynthetic gene murI of Streptococcus pneumoniae. Also provided are vectors and transformed heterologous host cells for expressing the MurI enzyme product and a method for identifying compounds that inhibit stem peptide biosynthesis.

Abstract: The invention provides human peptidyl-prolyl isomerases (HPPIP) and polynucleotides which identify and encode HPPIP. 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 HPPIP.

Abstract: Topoisomerase III polypeptides and DNA and RNA encoding such Topoisomerase III polypeptides and a procedure for producing such polypeptides by recombinant techniques is disclosed. Also disclosed are methods for utilizing such Topoisomerase III for the treatment of infection, particularly bacterial infections. Antagonists against such Topoisomerase III and their use as a therapeutic to treat infections, particularly bacterial infections are also disclosed. Also disclosed are diagnostic assays for detecting diseases related to the presence of Topoisomerase III nucleic acid sequences and the polypeptides in a host. Also disclosed are diagnostic assays for detecting polynucleotides encoding Staphylococcal Topoisomerase III and for detecting the polypeptide in a host.

Abstract: A human UDP galactose-4-epimerase polypeptide and DNA (RNA) encoding such polypeptide and a procedure for producing such polypeptide by recombinant techniques is disclosed. Also disclosed are methods for utilizing such polypeptide for the treatment of galactosemia. Also disclosed are diagnostic assays for the detection of mutations in the nucleic acid sequencing encoding human UDP galactose-4-epimerase protein and for detecting an altered level of human UDP galactose-4-epimerase protein in a sample derived from a host.

Abstract: The present invention provides an FK506 binding protein gene containing a base sequence coding for the amino acid sequence shown under SEQ ID NO:1, in particular such a FK506 binding protein gene containing the base sequence shown under SEQ ID NO:2, a method of producing a recombinant FK506 binding protein through expression of such gene, and the recombinant protein thus produced.The use of the gene of the present invention enables FK506 binding protein expression, and the protein is useful particularly in elucidating the mechanism of immunosuppression in living bodies or in developing or screening out therapeutic agents for autoimmune diseases (e.g. rheumatism, SLE, etc.), among others.

Abstract: The present invention provides a gene coding for maleate isomerase excellent in stability, a recombinant vector having the gene, and a transformant transformed with the recombinant vector. According to the present invention, maleate isomerase excellent in stability can be produced efficiently in a large amount.

Abstract: The invention provides YfiI pseudouridine synthase polypeptides and polynucleotides encoding YfiI pseudouridine synthase polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing YfiI pseudouridine synthase polypeptides to screen for antibacterial compounds.

Abstract: Hybrid polypeptides are provided formed with encapsulating regions from genes that encode for anabolic proteins. More particularly, the present invention relates to recombinant nucleic acid molecules that code for genes which encapsulate an attached protein within a matrix; preferably, these genes encapsulate a desired ("payload") polypeptide within starch, and more specifically within the starch granule matrix. Expression vectors comprising these recombinant nucleic acid molecules, and hosts therefor, and more specifically the starch-bearing portions of such hosts, transformed with such vectors, are also provided. Preferably, grain containing a foreign protein encapsulated within the starch is provided, useful to produce mammalian, fish and avian food. The invention also encompasses methods of producing purified protein from starch and particularly from starch granules, and industrial uses of such protein.

Abstract: Novel proteins of Flavobacterium sp. R1534 and the DNA sequences which encode these proteins are disclosed which provide an improved biosynthetic pathway from farnesyl pyrophosphate and isopentyl pyrophosphate to various carotenoid precursors and carotenoids, especially .beta.-carotene, lycopene, zeaxanthin and cantaxanthin.

Abstract: DNA sequences obtained from S. clavuligerus ATCC 27064, recombinant vectors incorporating such sequences and hosts transformed with such vectors are disclosed. The DNA comprises one or more genes coding for one or more enzymes involved in the biosynthesis of penicillin and cephalosporin .beta.-lactams and such enzymes are expressed by hosts into which the recombinant vectors are transformed. The DNA and the enzymes encoded thereby have utility in the preparation of penicillins and cephalosporins, both known and novel, possessing pharmacological, especially antimicrobial, activity.

Abstract: The present invention provides a human parvulin-like protein (HPVLP) and polynucleotides which identify and encode HPVLP. The invention also provides genetically engineered expression vectors and host cells comprising the nucleic acid sequences encoding HPVLP and a method for producing HPVLP. The invention also provides for agonists, antibodies, or antagonists specifically binding HPVLP, and their use, in the prevention and treatment of diseases associated with expression of HPVLP. Additionally, the invention provides for the use of antisense molecules to polynucleotides encoding HPVLP for the treatment of diseases associated with the expression of HPVLP. The invention also provides diagnostic assays which utilize the polynucleotide, or fragments or the complement thereof, and antibodies specifically binding HPVLP.

Abstract: An L-ribose isomerase which isomerizes aldoses such as L-ribose, D-lyxose, D-talose, D-mannose, L-allose and L-gulose into their corresponding ketoses such as L-ribulose, D-xylulose, D-tagatose, D-fructose, L-psicose and L-sorbose. The enzymatic reaction is a reversible equilibrium reaction. The L-ribose isomerase can be obtained from microorganisms of the genus Acinetobacter.

Abstract: The invention provides a human cyclophilin-type peptidyl-prolyl cis/trans isomerase (CPCI) and polynucleotides which identify and encode CPCI. 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 CPCI.

Abstract: The present invention provides a human parvulin-like protein (HPVLP) and polynucleotides which identify and encode HPVLP. The invention also provides genetically engineered expression vectors and host cells comprising the nucleic acid sequences encoding HPVLP and a method for producing HPVLP. The invention also provides for agonists, antibodies, or antagonists specifically binding HPVLP, and their use, in the prevention and treatment of diseases associated with expression of HPVLP. Additionally, the invention provides for the use of antisense molecules to polynucleotides encoding HPVLP for the treatment of diseases associated with the expression of HPVLP. The invention also provides diagnostic assays which utilize the polynucleotide, or fragments or the complement thereof, and antibodies specifically binding HPVLP.