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. The activity of an enzyme on a substrate can be improved by contacting the enzyme with a noble gas under specific temperatures and pressures. The preferred noble gases are krypton, neon, xenon, argon or mixtures of these gases. The enzymes whose activity can be improved are hydrolases, lyases, isomerases and ligases.

Abstract: A new homogeneous cytosolic binding protein (FKBP12.6), having a specific binding activity of about 4.8 mg FK-506 per mg protein and a molecular weight of about 10-12 kilodaltons, reversibly binds the immunosuppressant FK-506, but not cyclosporine A (CSA). The protein is unstable to heating at 56.degree. C. for 30 minutes losing its FK-506 binding affinity. The FKBP12.6 protein is isolated from the cytosol of mammalian tissues, preferably bovine or human brain tissue, and can be used in diagnostic and purification procedures involving FK-506-type macrolide immunosuppressants. The FKBP12.6 protein also has peptidyl-proline isomerase enzymatic activity, catalyzing the cis-trans isomerization of proline-containing peptide bonds. In addition, FKBP12.6 binds to and inhibits the phosphatase calcineurin in the presence of FK-506.

Abstract: A method for reacting an enzyme in a non-aqueous organic solvent is disclosed. The method comprises preparing a lyophilizate of a salt which activates the enzyme and an enzyme wherein the lyophilizate contains a weight ratio of salt to enzyme of at least 60% salt sufficient to activate the enzyme in an organic solvent. The method then calls for dispersion of the lyophilizate in a non-aqueous organic solvent in the presence of a substrate for the enzyme.

Abstract: A variety of chemical reactions are rate enhanced by the use of antibodies whose antibody binding sites are complementary to the reactant orientations or transition states which lead to the desired product. The reactions include pericyclic reactions such as Claisen and Cope rearrangements and Diels Alder reactions, peptide bond hydrolysis reactions, peptide fragment ligations, lactonizations and cyclic peptide syntheses, glycosylations, aldol additions, nucleoside syntheses and transesterification reactions. Haptens which are stable analogues of unstable transition states, or which are analogues of desired products, in some cases with leaving groups still attached for purposes of avoidance of product inhibition, are used to generate the antibodies.

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 shown in the specification.Also disclosed is a process for producing a carotenoid or a carotenoid related compound which is selected from the group consisting of geranylgeranyl 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 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 of 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 yeast PPIase characterized by possessing following properties: (1) acting on and isomerizing the bond X.sub.aa -Pro (wherein X.sub.aa stands for any amino acid and Pro stands for L-proline), (2) exhibiting a single molecular weight of about 17,000 daltons in the sodium dodecyl sulfate-polyacrylamide concentration gradient gel electrophoresis, (3) exhibiting a single isoelectric point of about 6.2 in the isoelectric focusing, and (4) inhibited by CsA;an E. coli PPIase-.beta. characterized by possessing the following properties: (1) acting on and isomerizing the bond X.sub.aa -Pro (wherein X.sub.aa stands for any amino acid and Pro for L-proline), (2) exhibiting a single molecular weight of about 20,000 daltons in the sodium dodecyl sulfate-polyacrylamide concentration gradient gel electrophoresis, (3) exhibiting a single isoelectric point of about 5.0 in the isoelectric focusing, and (4) no being inhibited by CsA;an E. coli PPIase-.alpha.

Abstract: A xylose isomerase gene from Thermus bacteria, such as Thermos aquaticus (ATCC 27634) and a gene having 60% or more of homology to the nucleotide sequence of Thermus aquaticus xylose isomerase gene of FIG. 1-3. A xylose isomerase from Thermus aquaticus characterized in that the xylose isomerase has the optimal pH of about 7, the stable pH range of from about 6 to 8.5, the optimal temperature of about 95.degree. C. and the molecular weight of about 44,000, and is stabilized with manganese or magnesium. A process for preparing a xylose isomerase comprising transforming a microorganism with a plasmid containing the above gene and a promoter, culturing the transformed microorganism and harvesting the produced xylose isomerase. A process for preparing fructose comprising isomerization of glucose to fructose in the presence of the above xylose isomerase.

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: The activity of a lipase is improved in processes where the lipase is contacted with a noble gas. The gases neon, argon, xenon, and krypton when contacted with the lipase improve the activity of the lipase when the process is performed at a pressure less than 100 atmospheres.

Abstract: A process for preparing a .sup.13 C-labelled compound which comprises reacting an alcohol .sup.13 C-labelled carbon source compound and a substrate in the presence of an enzyme system consisting of oxidase belonging to EC1 group and capable of converting the alcohol .sup.13 C-labelled carbon source compound to an aldehyde .sup.13 C-labelled carbon source compound, lyase belonging to EC4 group and capable of synthesizing a carbon-carbon bond and at least one of isomerases belonging to EC5 group and capable of isomerizing substrates to obtain a .sup.13 C-labelled compound specifically labelled with .sup.13 C in a specific carbon position.

Abstract: A yeast PPIase characterized by possessing the following properties: (1) acting on and isomerizing the bond X.sub.aa -Pro (wherein X.sub.aa stands for any amino acid and Pro stands for L-proline), (2) exhibiting a single molecular weight of about 17,000 daltons in the sodium dodecyl sulfate-polyacrylamide concentration gradient gel electrophoresis, (3) exhibiting a single isoelectric point of about 6.2 in the isoelectric focusing, and (4) inhibited by CsA;an E. coli PPIase-.beta. characterized by possessing the following properties: (1) acting on and isomerizing the bond X.sub.aa -Pro (wherein X.sub.aa stands for any amino acid and Pro for L-proline), (2) exhibiting a single molecular weight of about 20,000 daltons in the sodium dodecyl sulfate-polyacrylamide concentration gradient gel electrophoresis, (3) exhibiting a single isoelectric point of about 5.0 in the isoelectric focusing, and (4) no being inhibited by CsA;an E. coli PPIase-.alpha.

Abstract: A mannose isomerase having excellent properties for industrial use, such as high thermal stability and resistance to high substrate concentrations, can be produced by culturing a strain of Pseudomonas (sp. AM-9582), and extracting it from the cells of AM-9582. Mannose can be effectively produced from fructose of high concentrations using the enzyme.

Abstract: Process for manufacture of D-xylose characterized by the fact that:in a first step, syrup of D-xylulose is subjected to an enzymatic isomerization in M.sub.3 providing a mixture of D-xylose and D-xylulose,in a second step, the abovesaid mixture is subjected to chromatographic treatment in M.sub.4 leading to at least two fractions of which one is highly enriched in D-xylose (fraction X.sub.1) et of which the other is highly enriched in D-xylulose (fraction X.sub.2),in a third step, the fraction X.sub.2 is recycled through a pipe P to M.sub.3,the D-xylose being recovered from the fraction X.sub.1, the latter can also be subjected directly to a hydrogenation step.

Abstract: This invention is in the field of glucose isomerization enzymes. More specifically, the invention is directed to a novel xylose isomerase, a process for the preparation of this enzyme, the use of this enzyme in glucose isomerization processes, and glucose isomerization processes. The enzyme is preferably derived from Thermotoga maritima or Thermotoga neapolitana. The enzyme has a temperature optimum above 90.degree. C., pH optimum in the range of from 6 to 7 and a residual activity at 90.degree. C. of more than 40% after 30 minutes and/or residual activity at 98.degree. C. of more than 20% after 30 minutes. The enzyme can also be in immobilized form.

Abstract: A process for making L-aminoacylase includes a cultivation of microorganism selected from a specy of Alcaligenes, especially the Alcaligenes denitrificans DA 181, and a separation of a produced L-aminoacylase from the bacterial cells for obtaining the L-aminoacylase which may be further purified for the production of L-amino acid. The acylase made by such a process may have an increased stability, beneficial for its commercial and medical values.

Abstract: A mannose isomerase having excellent properties for industrial use, such as high thermal stability and resistance to high substrate concentrations, can be produced by culturing a strain of Pseudomonas (sp. AM-9582), and extracting it from the cells of AM-9582. Mannose can be effectively produced from fructose of high concentrations using the enzyme.

Abstract: A new homogeneous cytosolic binding (HCB) protein, having a specific binding activity of about 26 .mu.g FK-506 per mg protein and a molecular weight of about 10-12 kilodaltons, reversibly binds the immunosuppressant FK-506 but not cyclosporine A (CSA). The protein is stable to heating at 56 degrees C. for 30 minutes retaining its FK-506 binding affinity, and has the (partial) amino terminal amino acid sequence: H.sub.2 N-Gly-Val-Gln-Val-Glu-Thr-Ile-Ser-Pro- Gly-Asp-Gly-Arg-Thr-Phe-Pro-Lys- Ar g-Gly-Gln-Thr-X-Val-Val-His-Tyr-Thr-Gly-Met-Leu-Glu-Asp-Gly-Lys-Lys-Phe-As p (wherein X is undefined). The HCB protein is isolated from the cytosol of mammalian tissues, preferably human neoplastic T-cell lines, e.g., Jurkat, and can be used in diagnostic and purification procedures involving FK-506 macrolide type immunosuppressants. The HCB protein also catalyzes the cis-trans isomerization of proline-containing peptide bonds.

Abstract: A method for synthesizing camptothecin and camptothecin analogs using a novel hydroxyl-containing tricyclic intermediate and the camptothecin analogs produced by the process. The camptothecin analogs are effective inhibitors of topoisomerase I and show anti-leukemic and anti-tumor activity.

Abstract: The invention provides a novel bile acid sulfate sulfatase, a process for its preparation, and a method of assaying bile acid 3.alpha.-sulfates and total bile acids using the bile acid sulfatase.

Abstract: The invention provides a novel bile acid sulfate sulfatase, a process for its preparation, and a method of assaying bile acid 3.alpha.-sulfates and total bile acids using the bile acid sulfatase.

Abstract: A process for obtaining N-acetylneuraminic acid from N-acetylglucosamine is disclosed. The process is carried out in a reactor which contains both N-acylglucosamine-2-epimerase (E.C. 5.1.3.8) which isomerizes GlcNAc into ManNAc, and N-acetylneuraminic acid pyruvate lyase (E.C. 4.1.3.3) which catalyzes the reaction of the resulting ManNAc with pyruvic acid to give Neu5Ac. GlcNAc and Pyr are fed into the reactor and Neu5Ac is obtained in the outflow. The process is preferably carried out continuously and in particular in an enzyme membrane reactor at pH 7.5 and 25.degree. C., especially using residence times of 0.2 to 10 h, and with an excess of GlcNAc in comparison with Pyr which is subsequently added if necessary. Epimerase and lyase are preferably present in the reactor in a ratio of activities which is equivalent to the reciprocal value of the quotient of the conversion rates.

Abstract: An economical method of converting mannose to fructose uses a mannose isomerase from Pseudomonas cepacia immobilized on an alumina containing polyethyleneimine crosslinked with an excess of glutaraldehyde. The method utilizes mannose-containing aqueous solutions as the feedstock, and affords solutions in which at least 55% of the mannose has been converted to fructose. Because of the relatively higher levels of fructose than can be obtained by isomerizing glucose to fructose using glucose isomerase, substantial savings in separation of high fructose-containing products can be achieved. The process described represents the first economical mannose isomerase process.

Abstract: The present invention relates to acylamino acid racemase, production and use thereof.The acylamino acid racemase of the present invention racemizes optically active N-acyl-.alpha.-aminocarboxylic acid alone at pH values around the neutral level at a normal temperature under normal pressure in the presence of optical active amino acid; its use in combination with D- or L-aminoacylase enables the production of optically active D- or L-.alpha.-amino acid from DL-acyl-.alpha.-aminocarboxylic acid at a high level of efficiency.

Abstract: The invention relates to DNA sequences, recombinant DNA molecules and transformed host organisms, and to their use in a process for the genetic engineering preparation of a polypeptide having the biological activity of the enzyme mutarotase.

Abstract: A method for stabilization of extra-chromosomal elements in bacteria during cultivation which comprises transformation of a host bacterium having a defect in a chromosomal gene needed for the synthesis or maintenance of the cell envelope with an extra-chromosomal element capable of complementing the chromosomal gene defect of the host bacterium, the extra-chromosomal element also including an expressible DNA-sequence coding for a desired product.

Abstract: A method of culturing an amino acid racemaseproducing microorganism of the genus Pseudomonas, which comprising culturing said microorganism in a culture medium containing at least one compound selected from the group consisting of glycerol, ethanol, tartaric acid, fumaric acid and succinic acid as a carbon source, and recovering microorganism cells containing the amino acid racemase in an increased amount.

Abstract: The invention relates to a process for the microbiological preparation of aldose-1-epimerase by cultivating microorganisms in a nutrient medium and release of the enzyme from the cells, which process is characterized in that microorganisms of the genus Acinetobacter are used.

Abstract: Proteins having chorismate mutase-prephenate dehydratase (CMPD) activity, but lacking phenylalanine sensitivity are produced by genetic engineering. The proteins contain a sequence substantially corresponding to the N-terminal 337 amino acids of Escherichia coli CMPD. Expression vectors including genes coding for those proteins and regulatory DNA enabling their expression are used to transform host microorganisms, which are cultured to produce phenylalanine.

Abstract: A process is disclosed for chemically modifying naturally occurring proteins to produce enzyme-like modified proteins. The process comprises partially denaturing a cofactor containing holoprotein by removal of the cofactor to produce a partially denatured cofactorless or so-called apoprotein. The partially denatured protein is contacted with an inhibitor of a selected model enzyme and cross-linked. The resultant protein product is an enzyme-like modified protein having the catalytic characteristics of the model enzyme whose inhibitor is contacted with the partially denatured apoprotein.

Abstract: A process for preparing uridine diphosphate-N-acetylgalactosamine, which comprises treating a reaction solution obtained by the enzymatic conversion of uridine diphosphate-N-acetylglucosamine to uridine diphosphate-N-acetylgalactosamine, with uridine diphosphate-N-acetylglucosamine pyrophosphorylase to decompose the remaining uridine diphosphate-N-acetylglucosamine in the solution and then separating therefrom the uridine diphosphate-N-acetylgalactosamine for purification. In one aspect of this invention, it relates to a method for measuring the activity of .alpha.-N-acetylgalactosaminyl transferase characterized by the use of said reaction solution as the substrate for the transferase.

Abstract: Disclosed is a process for purifying an enzyme contained in a solution such as cell extract liquor or fermentation culture liquor. The crude enzyme solution is brought into contact with either a strongly acidic cation exchange resin of high porous type or a strongly basic anion exchange resin of high porous type to adsorb the enzyme on the resin. An eluting agent is then passed through the resin to elute out the enzyme as a purified enzyme solution.

Abstract: Spherically shaped bacterial cell aggregates are produced by spheronizing extruded flocculated cells. The cells are flocculated from aqueous medium with a cross-linked polyamine which is the reaction product of an epihalohydrin/polyamine copolymer and a cross-linking agent. Prior to being extruded, a filter cake having 68 to 76 weight percent water is produced by filtration of the flocculated cells, and the filter cake is ground into particles no greater than 60 mesh. Spheronizing is with a plate rotating at a tangential velocity of 4.5 to 12 meters per second within a cylinder containing the plate. Toughness of the spherical aggregates produced can be increased by the addition of a binder after filtration and before extrusion. During spheronizing, fines may be produced. These fines can be recycled by mixing them with the wet filter cake and binder before extrusion.

Abstract: Process for preparing L-fructose from L-mannose by contacting L-mannose with L-mannose isomerase produced by a mutant microorganism selected from the group consisting of the genera Escherichia, Lactobacillus, and Klebsiella cultivated in the absence of an inducing sugar.

Abstract: Sucrose mutase in whole cells of Protaminobacter rubrum #l is immobilized by contacting the cells with tannic acid, polyethylenimine, and an adduct of glutaraldehyde and an epihalohydrin/polyamine copolymer. The resultant reaction product has improved sucrose mutase activity and physical characteristics for use in a packed-bed reactor to convert sucrose to palatinose.

Abstract: Whole microbial cells containing enzymes not sensitive to glutaraldehyde such as sucrose mutase or glucose isomerase are immobilized by forming a reaction product of the cells in an aqueous medium with glutaraldehyde, reacting the reaction product with polyethylenimine to flocculate the reaction product, and recovering the flocculated reaction product from the aqueous medium. Reacting the cells with glutaraldehyde prior to reacting with polyethylenimine results in flocculated cells that can be more easily separated from the aqueous medium.

Abstract: A process for the preparation of L-tryptophan comprises reacting indole with serine in the presence of tryptophan synthetase or tryptophanase, wherein DL-serine or D-serine is used and a serine racemizing enzyme is included in the reaction system and reacted with the serine.

Abstract: The invention relates to a novel enzymatic labelling agent for immunoenzymatic assays. The labelling agent is .DELTA..sub.5, 3-keto-steroid isomerase. The enzymatic activity bound to the antibody can be revealed advantageously by: (1) direct reading on the supernatent liquor or on the cake obtained by precipitation by means of a solvent, such as polyethylene glycol, with the use of soluble antibodies; (2) reading in solution after liberation of the enzyme from the antigen-antibody complex by a reducing agent, such as dithiotreitol; (3) immediate reading after filtration of a substrate solution on the insolubilized antibody-antigen-enzyme complex. The method can be used for immunoenzymatic assays automatically on substances present in very low concentrations, such as hypophysial hormones and plasmatic steroids.

Abstract: A shaped bacterial cell aggregate having increased hardness is produced by adding previously-produced dried finely-divided bacterial cell aggregate to bacterial cell aggregate subsequent to its formation but prior to its shaping.