Abstract: Methods are disclosed which employ cold-shock proteins, such as Trigger Factor, to confer cold-tolerance to cells and to enhance viability of cells at temperatures at which the cells would not normally remain viable. Proteins having peptidyl-proline isomerase activity are also disclosed as being useful to confer cold-tolerance to cells. Cells which overexpress cold-shock proteins are useful in cell-based expression systems which are intended to express proteins at low temperatures.

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: 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: A novel topoisomerase IV, nucleotide sequences coding for said enzyme, corresponding vectors, and the use of said enzyme for screening biologically active materials.

Abstract: The present invention provides a polynucleotide (pdih) the partial sequence for which was initially isolated from a lung cDNA library and which identifies and encodes a novel human protein disulfide isomerase (PDIH). The invention provides for genetically engineered expression vectors and host cells comprising the nucleic acid sequence encoding PDIH. The invention also provides for the use of purified PDIH and its agonists in the commercial production of recombinant proteins and in pharmaceutical compositions for the treatment of diseases associated with the abnormal expression of PDIH. Additionally, the invention provides for the use of antisense molecules to pdih or inhibitors of PDIH in pharmaceutical compositions for treatment of diseases.

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: A novel polypeptide, acylglucosamine 2-epimerase as shown in FIG. 1 and derivatives thereof, DNA coding for said enzyme, a recombinant vector containing said enzyme, a transformant integrated thereinto said vector, a method for producing said enzyme and a method for producing N-acetylmannosamine and N-acetylneuraminic acid using renin binding protein.

Abstract: The invention relates to sucrose isomerases, to DNA sequences coding therefor, and to novel processes for the production of non-cariogenic sugars.

Abstract: The invention relates to sucrose isomerases, to DNA sequences coding therefor, and to novel processes for producing noncariogenic sugars.

Abstract: Cyclophilin-60 proteins, nucleic acids, and monoclonal antibodies are provided.

Abstract: Disclosed is a human is a hTopI-.alpha. polypeptide and DNA (RNA) encoding such hTopI-.alpha. polypeptide. Also provided is a procedure for producing such polypeptide by recombinant techniques and antibodies and antagonists against such polypeptide. Also provided are methods of using the antibodies and antagonist inhibitors to inhibit the action of hTopI-.alpha. for therapeutic purposes such as an antitumor agent, to detect an autoimmune disease, or retroviral infections and to treat adenocarcinoma of the colon. Diagnostic methods for detecting mutations in the coding sequence and alterations in the concentration of the polypeptides in a sample derived from a host are also disclosed.

Abstract: A protein derived from a strain of methylotrophic yeast which has a protein disulfide isomerase activity having the amino acid sequence as set forth in SEQ ID No. 1, or protein in which the amino acid sequence has been modified by deletion or addition of one or a few amino acids, or substitution with other amino acid(s) and which has a protein disulfide isomerase activity; and a process for production thereof.

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 III and for detecting the polypeptide in a host.

Abstract: The invention provides a method and expression system for enhancing secretion of hyperproduced homologous and heterologous exoproteins in gram-positive bacteria such as Bacillus sp. The method and system comprise overproduction of PrsA protein in a gram-positive bacterial host also overproducing at least one exoprotein of interest. Use of the method and system of the invention results in greatly enhanced secretion of the synthesized exoproteins into the growth medium. Once in the growth medium these secreted exoproteins can be recovered and purified in a straightforward manner.

Abstract: The present invention relates xylose isomerases obtainable from strains of Thermotoga. More specifically the invention provides DNA sequences encoding xylose isomerases, expression vectors and host cells comprising the DNA sequences of the invention, and methods of producing xylose isomerases. The invention also provides isolated xylose isomerases free from homologous impurities and obtained by the method of the invention.

Abstract: A fungal protein disulfide isomerase obtainable from fungal species belonging to Aspergillus, especially A. oryzae, or A. niger, is disclosed. Furthermore sequences for recombinant production of the protein disulfide isomerase are disclosed.

Abstract: Protein disulfide isomerase, which catalyzes the exchange reaction between sulfhydryl and disulfide in a protein for formation of the most stable natural type disulfide bonds, is useful for formation of natural type disulfide bonds in a protein which is produced in a prokaryotic cell.

Abstract: An expression cassette, operable in a recombinant host, comprising a heterologous DNA coding sequence encoding a protein, which is functional, alone or in cooperation with one or more additional proteins, of catalyzing an oxidation step in the biological pathway for conversion of cholesterol into hydrocortisone, which step is selected from the group consisting of:the conversion of cholesterol to pregnenolone;the conversion of pregnenolone to progesterone;the conversion of progesterone to 17.alpha.-hydroxy-progesterone;the conversion of 17.alpha.-hydroxyprogesterone to cortexolone;the conversion of cortexolone to hydrocortisone, and thecorresponding control sequences effective in said host.

Abstract: Mutant proteins of human DNA topoisomerase I having an amino acid sequence in which tyrosine at the 592nd position of human DNA topoisomerase I is lacking or replaced with phenylalanine and which contains at least 30 amino acids in succession of the amino acid sequence subsequent to the 542nd amino acid of human DNA topoisomerase I.Above described mutant proteins react with anti-Scl-70, antibody in the sera of autoimmune disease patients of diffuse scleroderma and can be produced by genetic engineering using E. coli, and therefore they are useful as a diagnostic agent of scleroderma.

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 the use of certain chemical compounds which interfere with the biosynthesis of cholesterol and medicaments comprising such compounds for stimulating the meiosis of oocytes and spermatozoon in vivo, ex vivo and in vitro.

Abstract: The invention relates to novel microorganisms, their use and method of producing L-.alpha.-amino acids. In particular, microorganisms DSM 7329 and 7330 are suitable for the production of L-.alpha.-amino acids from corresponding hydantoins or carbamoyl-.alpha.-amino acids. These novel microorganisms are simple to cultivate and make possible high L-.alpha.-amino acid yields from different substrates.

Abstract: The present invention relates to the use of these cyclophilins, hereinafter referred to as "cyclophilin-like proteins (CLP)" in a method for identifying compounds capable of binding to and/or inhibiting the enzymatic activity of these proteins. Such compounds may be further screened for their ability to inhibit parasites which are not susceptible to the anti-parasitic effects of CsA.

Abstract: The present invention clarifies the primary structure of human-originated PGIS and the nucleotide sequence encoding same. The PGIS and its DNA are useful as reagents for the development of therapeutic agents for the cardiovascular diseases induced by the production imbalance between PGI.sub.2 and TXA.sub.2, and as diagnostics for determining the in vivo tissue expression level and distribution of PGIS or mRNA thereof. Moreover, they can be used as therapeutic agents for cardiovascular diseases, which introduce PGIS and the like into human or other animals in a lesion-specific manner. The production method of the present invention is useful for the easy and efficient mass production of the human-originated PGIS. The antibody of the present invention is useful for the purification of the human-originated PGIS and immunohistochemical analysis of the cause of a disease.

Abstract: L-Ketohexoses such as L-fructose and L-tagatose are produced from L-psicose and L-sorbose as substrates by allowing D-ketohexose 3-epimerase to act on these substrates. The yield and the production cost of the produced L-fructose and L-tagatose are industrially acceptable. The enzyme is preparable from microorganisms, e.g. those of the genus Pseudomonas.

Abstract: An oxido-reducto synthesis is carried out in a liquid medium with enzymatic transformation of a substrate with an oxido-reducto cofactor dependent enzyme in the presence of a cofactor for the transformation; the enzyme is immobilized by a support which is stationarily disposed in a flow path along which a liquid medium containing the substrate flows in contact with the support.

Abstract: The present invention provides a polynucleotide (pdih) the partial sequence for which was initially isolated from a lung cDNA library and which identifies and encodes a novel human protein disulfide isomerase (PDIH). The invention provides for genetically engineered expression vectors and host cells comprising the nucleic acid sequence encoding PDIH. The invention also provides for the use of purified PDIH and its agonists in the commercial production of recombinant proteins and in pharmaceutical compositions for the treatment of diseases associated with the abnormal expression of PDIH. Additionally, the invention provides for the use of antisense molecules to pdih or inhibitors of PDIH in pharmaceutical compositions for treatment of diseases resulting secretion of PDIH.

Abstract: A novel polypeptide, acylglucosamine 2-epimerase as shown in FIG. 1 and derivatives thereof, DNA coding for said enzyme, a recombinant vector containing said enzyme, a transformant integrated thereinto said vector, a method for producing said enzyme and a mrthod for producing N-acetylmannosamine and N-acetylneuraminic acid using renin binding protein.

Abstract: The present invention provides a method for identifying or selecting from a population of eukaryotic cells cultivated on or in a medium containing at least one compound, cells which have a metabolic advantage as a result of having being transformed, wherein:i) the cells are transformed with a nucleotide sequence or a co-introduced nucleotide sequence one of which comprises a region which: (a) encodes a protein which is involved in the metabolism of the compound, and/or (b) regulates the activity of the protein; andii) the compound is mannose or xylose or a derivative or a precursor of these, or a substrate of the protein, or is capable of being metabolized by the transformed cells into such a substrate, with the proviso that the compound is not mannose when the protein is mannose 6 phosphate isomerase.

Abstract: An FK506 binding protein of mammalian origin of approximate size (M.sub.r) 52,000, isolated by FK506 affinity chromatography and a corresponding human cDNA of approximate size 2.2 Kb, isolated by screening a human placenta cDNA library with a DNA probe whose sequence predicts a consensus amino acid sequence present in five FKBP12 sequences and in the human FKBP13 sequence.

Abstract: L-aspartic acid is produced by repeating the following respective steps:(1) a reaction step of producing ammonium L-aspartate from an aqueous solution containing monoammonium maleate in accordance with an isomerization reaction and an enzyme reaction caused by aspartase in the presence of ammonia;(2) an ammonia-eliminating step of converting substantially all produced ammonium L-aspartate into monoammonium salt by distilling or stripping a reaction solution obtained in the step (1);(3) a crystallization step of crystallizing L-aspartic acid and producing monoammonium maleate from a solution obtained in the step (2) by adding maleic acid, maleic anhydride or both;(4) a solid-liquid separation step of separating L-aspartic acid crystals precipitated in the step (3) from a mother liquor containing monoammonium maleate; and(5) a recycle step of supplying the mother liquor containing monoammonium maleate obtained in the step (4) to the step (1) to be used as a raw material for the reaction.

Abstract: Disclosed is a human is a hTopI-.alpha. polypeptide and DNA (RNA) encoding such hTopI-.alpha. polypeptide. Also provided is a procedure for producing such polypeptide by recombinant techniques and antibodies and antagonists against such polypeptide. Also provided are methods of using the antibodies and antagonist inhibitors to inhibit the action of hTopI-.alpha. for therapeutic purposes such as an antitumor agent, to detect an autoimmune disease, or retroviral infections and to treat adenocarcinoma of the colon. Diagnostic methods for detecting mutations in the coding sequence and alterations in the concentration of the polypeptides in a sample derived from a host are also disclosed.

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: A glucosamine 6-phosphate deaminase having specific physicochemical properties. A process for producing the glucosamine 6-phosphate deaminase which comprises incubating a microorganism belonging to the genus Vibrio and harvesting the glucosamine 6-phosphate deaminase from the culture thus obtained.

Abstract: Protein disulfide isomerase, which catalyzes the exchange reaction between sulfhydryl and disulfide in a protein for formation of the most stable natural type disulfide bonds, is useful for formation of natural type disulfide bonds in a protein which is produced in a prokaryotic cell.

Abstract: A highly thermostable polypeptide possessing protein disulfide isomerase (PDI) activity, a gene coding for the polypeptide and a process for producing the polyeptide are provided. The polypeptide possessing PDI activity is characterized by A) having a capability of catalyzing a disulfide exchange in proteins, B) recognizing mainly ribonuclease A as a substrate, C) having a suitable active temperature of 20.degree. to 70.degree. C., D) being stable at a pH value of 6 to 9, and E) having a molecular weight of about 60,000 to 62,000. Since it has a higher thermostability and exhibits a stable activity in a wider dithiothreitol concentration range as compared with the conventional PDI, it is possible to provide a novel enzyme active protein which can be advantageously used for a refolding reaction of certain proteins.

Abstract: D-ketohexose 3-epimerase, a novel epimerase, is obtained by cultivating bacteria of the genus Pseudomonas including Pseudomonas cichorii ST-24 (FERM BP-2736). D-Ketohexose 3-epimerase epimerizes D-ketohexose, D-ketopentose and L-ketopentose at their C-3 positions to form their corresponding epimeric counterparts in a high yield at a high conversion rate. Interconversion reaction using D-Ketohexose 3-epimerase yields mixture of intact ketose and its epimeric counterpart which can impart an appropriate sweetness, gloss and improve taste quality when used in foods, beverages, feeds, pet foods, dentifrice, cachou, sublingual agents and internal medicines.

Abstract: Fumaric acid is produced by reacting maleic acid in an aqueous solution with a microorganism which has maleate isomerase activity or with a preparation from the microorganism having the maleate isomerase activity, and producing fumaric acid in a reaction solution by enzymatic isomerization of maleic acid carried out under the condition that the dissolved oxygen concentration in the reaction solution is substantially maintained at 4 ppm or less, for example, by sealing the reaction solution with one or more gases selected from N.sub.2, Ar, and He.

Abstract: An enzyme reaction stabilizer consisting of poly-L-lysine or its salt as an effective constituent, a method for using the enzyme reaction stabilizer, and an enzyme preservative consisting of poly-L-lysine or its salt as an effective constituent are disclosed in this invention. While enzyme reactions are carried out, the enzyme reaction stabilizer and the method for using it prevents both the deactivation of enzymes due to the proliferation of included microorganisms and the decomposition of the reaction products and there is an advantage in that the enzyme reaction stabilizer can easily be separated from the reaction products. On the other hand, while the enzyme solution is preserved, the enzyme preservative of this invention prevents the deactivation of the enzyme due to the proliferation of included microorganisms in the enzyme solution or the enzyme preserving solution.

Abstract: A type 1 topoisomerase, designated topoisomerase V, has been isolated and substantially purified from the halophilic thermophilic methanogen bacterium Methanopyrus kandleri. The topoisomerase was purified by a process including the steps of lysing cells of M. kandleri to form a lysate, treating the lysate with polyethyleneimine to form a precipitate and a supernatant, precipitating the polyethyleneimine supernatant with ammonium sulfate, chromatographing the ammonium sulfate precipitate on phosphocellulose to produce a phosphocellulose eluate, chromatographing the phosphocellulose eluate on heparin to produce a heparin eluate, and chromatographing the heparin eluate on a column capable separating proteins by molecular size therein to produce a substantially purified thermostable DNA topoisomerase V. Topoisomerase V can relax DNA and can unlink DNA by reducing the linking number of closed circular DNA.

Abstract: A method for producing a fusion peptide. A vector is provided with nucleic acid encoding a carrier peptide and at the 3' end of the nucleic acid, a unidirectional restriction endonuclease cleavage site recognized by a restriction endonuclease with the ability to create a non-palindromic 3-base overhang. The vector is cleaved with the restriction endonuclease to produce a cleaved vector. One or more nucleic acids encoding a desired peptide and having at least a 3-base overhang at each end configured and arranged for ligation with the cleaved vector is then ligated to the cleavage site.

Abstract: Disclosed are DNA sequences which are useful for the synthesis of carotenoids such as lycopene, .beta.-carotene, zeaxanthin or zeaxanthin-diglucoside, that is, DNA sequences encoding carotenoid biosynthesis enzymes. These DNA sequences are the sequences 1- 6, respectively, shown in the specification.Also disclosed is a process for producing a carotenoid compound which is selected from the group consisting of prephytoene pyrophosphate, phytoene, lycopene, .beta.-carotene, zeaxanthin and zeaxanthin-diglucoside, which comprises transforming a host with at least one of the DNA sequences 1- 6 described above and culturing the transformant.

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: The present invention relates to a DNA fragment containing a gene encoding acylamino acid racemase. The acylamino acid racemase can be produced industrially at high efficiency using the DNA fragment.

Abstract: A polypeptide having mutarotase activity is obtained from a host microorganism that has been transformed with a molecule having a recombinant DNA sequence. The molecule having a recombinant DNA sequence is prepared by removing the first 60 nucleotides of a DNA sequence originating from the genome of Acinetobacter calcoaceticus that codes for the polypeptide, modifying the following 21 nucleotides and fusing of the resultant structural gene with the start of the tetracycline-repressor gene and with an effective promotor sequence. The tetracycline-regressor gene and the promotor sequence preferably originate from the same microorganism such as E. coli in which expression of the polypeptide is carried out, and result in increased yield of the expressed polypeptide having mutarotase activity.

Abstract: A purified preparation of FKBP-13, an FK506- and rapamycin-binding immunophilin; fragments and chimeras of FKBP-13 capable of binding FK506 and/or rapamycin; and DNA molecules encoding such polypeptides.

Abstract: A highly thermostable polypeptide possessing protein disulfide isomerase (PDI) activity, a gene coding for the polypeptide and a process for producing the polypeptide are provided. The polypeptide possessing PDI activity is characterized by A) having a capability of catalyzing a disulfide exchange in proteins, B) recognizing mainly ribonuclease A as a substrate, C) having a suitable active temperature of 20.degree. to 70.degree. C., D) being stable at a pH value of 6 to 9, and E) having a molecular weight of about 60,000 to 62,000. Since it has a higher thermostability and exhibits a stable activity in a wider dithiothreitol concentration range as compared with the conventional PDI, it is possible to provide a novel enzyme active protein which can be advantageously used for a refolding reaction of certain proteins.

Abstract: The present invention relates to the use of these cyclophilins, hereinafter referred to as "cyclophilin-like proteins (CLP)", in a method for identifying compounds capable of binding to and/or inhibiting the enzymatic activity of these proteins. Such compounds may be further screened for their ability to inhibit parasites which are not susceptible to the anti-parasitic effects of CsA.

Abstract: A process for determining the concentration of cyclosporine in a sample, which comprises adding to the sample a predetermined amount of an isomerase which isomerizes N-terminal peptide to proline, then measuring the enzyme-catalyzed inhibition of the isomerization of a proline-containing substrate, and determining the concentration or cyclosporine from a calibration curve.

Abstract: A structural gene encoding a polypeptide with .DELTA.8-7 sterol isomerase activity is disclosed, Recombinant DNA molecules useful for transforming yeast and mutant yeast transformed with such recombinant DNA molecules are also disclosed. The structural gene is useful for modulating the accumulation of sterols in yeast comprising increasing the expression level of a structural gene encoding a polypeptide having .DELTA.8-7 sterol isomerase activity in a mutant yeast having single or double defects in the expression of sterol biosynthetic enzymes is provided.