Abstract: This invention relates to a process of preparation of nucleotide polymers wherein a lysate of a bacterial culture is passed successively through three columns (ion exchange resin, hydrophobic resin and molecular sieve), whereby there is obtained a substantially pure polynucleotidephosphorylase solution.Polymerization of this agent leads to non toxic and non pyrogen products.This invention relates also to products thus obtained and to therapeutic compositions containing the same.

Abstract: A method for enzyme immunoassay includes contacting under binding conditions a liquid suspected of containing an analyte, an antianalyte affixed to a solid support and a tracer having an enzyme conjugated thereto. A bound fraction is separated from the liquid and incubated in a second liquid with a masked ligand. The masked ligand is converted by the enzyme on the bound fraction to give free lignad which binds to an antiligand. A signal system, such as a signal enzyme and substrate therefor, or a label-loaded vesicle and vesicle lysing agent, is added to generate a signal used to detect or measure the analyte in the liquid. The invention includes a kit of materials useful in performing the assay of the invention.

Abstract: For selectively and reversibly adsorbing tPA, pro-tPA or mixtures thereof, from an aqueous medium, for example, derived from a melanoma cell culture, the aqueous medium is intimately contacted with an affinity reagent comprising an immobilized Kunitz-type inhibitor, substantially as occurs in and is extractable from seeds of an Erythrina species, e.g., Erythrina latissima, having Erythrina species, the following combination of characteristics:(a) it is an inhibitor of the Kunitz-type;(b) it inhibits trypsin;(c) it inhibits plasmin;(d) it has no effect on urokinase;(e) it inhibits tPA; and(f) in its immobilized form, it is a selected adsorbent for tPA and its precursor pro-tPA, to thereby cause the tPA, pro-tPA or mixtures thereof to become selectively adsorbed on the affinity reagent.

Abstract: A mixture containing a single-chain tissue plasminogen activator (tPA) and/or double-chain tPA is brought into close contact with a column carrying an immobilized Erythrina trypsin inhibitor as an affinity agent. Adsorbed protiens are eluted with eluents having different pHs with or without arginine or benzamidine, so that single-chain tPA is obtained in the eluent with a pH at least 4.5 and double-chain tPA is obtained in the eluent with a pH lower than 4.5.

Abstract: A method of protecting soluble proteins such that their biological activity is preserved after freezing by exposing the protein to an amino acid or trimethylamine-N-oxide and transition metal ion prior to freezing. The protected protein can then be thawed without denaturation or impairment of the protein's biological activity. The protein is preferably exposed to the amino acid or trimethylamine-N-oxide by placing it in a 25-100 mM aqueous solution of organic solute and 1 mM Zn.sup.+2. This method is especially effective in preserving the biological activity of fragile proteins such as the enzyme phosphofructokinase. The present method can be used to preserve pharmaceutically useful proteins in a frozen form for storage and distribution. The treated protein can be thawed and administered directly to a user without removing the cryoprotectant since the amino acid or oxide and trace amounts of many transition metal ions are nontoxic.

Abstract: This invention relates to dipeptidase of lactic acid bacteria in isolated form, characterized by a molecular weight of 55 kD.+-.5 kD; an isoelectric point of 4.4.+-.0.4; a substrate range including the dipeptides leu-leu, leu-met, leu-val, leu-gly, val-leu, phe-leu and ala-ala, but not the dipeptides his-leu, .gamma.-glu-leu, gly-leu, .alpha.-glu-ala and peptides of 3 or more amino acid residues; and a hydrolyzing activity with a temperature optimum at 50.degree. C..+-.10.degree. C., a pH optimum at 8.+-.1, and sensitivity to the metal chelating EDTA and to the reducing agents dithiotreitol and mercaptoethanol.

Abstract: Thiol gels are prepared from hydroxyl containing polymers by forming a 2-fluoro-1-methylpyridoxy derivative of the polymer followed by reacting the derivative with sodium dimethyldithiocarbamate and reducing to produce a sulfhydryl substituted polymer. Alternatively, the derivative is reacted with dithiothreitol to produce a sulfhydryl substituted polymer having free sulfhydryl groups spaced from the polymer. The thiol gel can be activated by means of 2,2'-dipyridyl disulfide and reacted with a free sulfhydryl containing biologically active ligand in order to provide an insolubilized ligand which can be used for various purposes including use as an immobilized biologically active material or use as a matrix in covalent chromatography.

Abstract: A method for producing glucose at a high yield includes the step of saccharifying liquefied starch with a glucoamylase in the presence of a sugar transferase capable of the transfer reaction with a .alpha.-1,4-glucosidic bond.

Abstract: This invention discloses a fibrinophilic urokinase complex which is a complex of urokinase with a urokinase inhibitor or tissue activator inhibitor and has a molecular weight of 97,500.+-.3,000 and a method for the production of this fibrinophilic urokinase complex from urine. The urokinase complex can be used, as a thrombolytic agent exhibiting an outstanding ability to dissolve thrombus.

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: Human cell lines that proliferate in the absence of serum or of any macromolecular growth factors are produced by a process comprising removing serum containing culture medium from the culture of a human cell line that requires serum or macromolecular growth factors for growth, replacing the serum-containing culture medium with culture medium deprived of serum and of any macromolecular growth factors, feeding the adherent or non-adherent cells with serum-free and growth factor-free culture medium using conventional cell culture conditions, culturing the cells until they propagate reproducibly and indefinitely in the absence of serum and of any macromolecular growth factors and a serum and growth factor independent cell line has been established. The novel cell lines can be used for the production of biologically active compounds such as proteins, especially plasminogen activators.

Abstract: Two components, trypsin and kallidinogenase, in human urine are concentrated simultaneously by allowing human urine at neutral pH, collecting bubbles thus formed to obtain the concentrate of the two components, adjusting the concentrate to weak acidity, contacting the acidified concentrate with chitosan to allow the two components to be adsorbed onto chitosan, eluting the components from the adsorbent with aqueous ammonia solution, and neutralizing and heating the eluate at about 60.degree. C. for about 10 hours to make the eluate virus-free, followed by separating the components from the eluate.

Abstract: Deacetoxycephalosporin C synthetase is provided in purified form via chromatography of crude cell-free extracts over a weak anion exchange resin followed by gel filtration and hydroxylapatite chromatography, all carried out in the presence of glycerol, a C.sub.1 -C.sub.3 alkyl monohydric alcohol, e.g., ethanol, a sulfhydryl containing reducing agent, e.g., dithiothreitol, and ascorbate. The purified enzyme which possesses both expandase and hydroxylase activities can be further purified by chromatography over a strong anion exchange resin.

Abstract: A method for the isolation and quantitative detection of a selected single-stranded target polynucleotide from solution. The target polynucleotide is hybridized in solution to a single-stranded mediator polynucleotide, a probe polynucleotide, and an immobilized polynucleotide sequence. The sequence of the mediator polynucleotide comprises a first sequence complementary to a first portion of the target polynucleotide sequence and a second nucleotide sequence complementary to a portion of a single-stranded immobilized polynucleotide sequence. The probe polynucleotide, which carries a detectable label, is complementary to a second portion of the necleotide sequence of the target. The immobilized polynucleotide is immobilized by attachment to a solid support, and, through hybridization to the mediator polynucleotide, functions to immobilize the entire immobilized polynucleotide/target polynucleotide/probe polynucleotide "sandwich".

Abstract: The invention relates to a process for the separation of biotechnologically produced valuable materials form a cell suspension by crossflow microfiltration to obtain an high specific permeate flux while retention stays near 0% for long periods. To enable crossflow microfiltration to be carried out on an industrial scale in biotechnology in the separation of biotechnologically produced extracellular valuable materials from a cell suspension, particularly in the separation of alkaline protease for recovering enzyme, under economically acceptacle conditions, alkaline protease of relatively high molecular weight, more especially enzyme >20,000 daltons, is separated from a fermenter solution using polysulfone tubes having micropore diameters of from 0.3 to 0.5 .mu.

Abstract: An enzyme preparation having specific bilirubin degrading activity is described. It is isolated from plants (e.g. artichokes) of the Compositae family. It exhibits higher specificity towards bilirubin and has higher specific activity (i.e. turnover number of moles of substrate per minute per mg of protein) than similar enzymes isolated from other sources. Assay compositions, analytical elements and methods for use of such are also described.

Abstract: A sugar syrup is prepared by saccharifying a liquefied starch hydrolyzate at a pH of about 4 to about 5.5 and a temperature above 55.degree. C. with a heat stable, aciduric, pullulanase obtained from rice and an .alpha.-1,4 carbohydrase. The pullulanase employed is substantially free of maltase and transglucosidase activity. In one embodiment, a dextrose syrup is prepared by saccharifying a thinned starch hydrolyzate with glucoamylase and the rice pullulanase. Maltose syrup is prepared using the rice pullulanase and a maltogenic enzyme.

Abstract: Extracellular enzymes can be recovered from a whole fermentation beer by adding to the whole beer a mixture of a polymer and an inorganic salt. This total mixture produces an enzyme-rich polymer phase and a cell debris-containing, enzyme-poor salt phase which can be separated to produce an enzyme-rich product from the polymer phase.

Abstract: The separation of xylanases from mixtures thereof with other hemicellulases, particulary cellulase produced by the culturing of hemicellulolytic microorganisms, particularly the fungus Trichoderma harzianum E58 and Trichoderma reesei by ultrafiltration through an ultrafiltration membrane having a low molecular weight cut-off point in the range of about 1,000 to 20,000 daltons to obtain a cellulase rich retentate and xylanase rich ultrafiltrate. The dilute xylanase rich filtrate from the ultrafiltration is concentrated and purified by adsorption and elution from an insoluble matrix, e.g. a cationic exchange resin. The xylanase obtained is suitable for use in the hydrolysis of hemicellulose for which it is selective, particularly in the presence of cellulose and the cellulase rich retentate is suitable for the hydrolysis of cellulose.

Abstract: A fluid starting material based on white of egg is subjected to at least one microfiltration by bringing the said material into contact, under pressure and in tangential flow, with a membrane capable of separating off the particles having a size of between about 0.02 micron and 10 microns, giving, firstly, a microfiltrate which passes through the membrane and contains at least a fraction of the lysozyme of the starting material and, secondly, a product held back by the membrane, which retains the natural properties of the starting material based on white of egg but is depleted in lysozyme. The obtained lysozyme can be conventionally used in particular in cheese-making.

Abstract: Disclosed is a method for causing the dissociation of microbial mycelium and extracellular lipase bound thereto and increasing the measurable activity of the lipase. The method involves treating an aqueous suspension of the mycelium with the anhydride of a dicarboxylic acid which results in dissociation of the mycelium and lipase thereby facilitating recovery of the lipase.

Abstract: Porous glass beads for filtation applications having a homogeneous metaloxane structure and comprising oxides of Si, Zr and optionally Ti and Al. A preferred method for making these beads comprises the steps of (a) providing a mother solution of Si and Zr alkoxides in a water soluble solvent, for instance a lower aliphatic alcohol, (b) providing a liquid dispersant phase in which solution (a) is dispersible and stirring this liquid phase sufficiently to cause (a) to be formed into droplets of substantially uniform size when added to (b), (c) adding (a) to (b) at a rate sufficient to provide said droplets and effecting the hydrolysis of the alkoxides contained therein with consecutive gelation of said droplets into corresponding hardened beads of condensed mixed Si and Zr hydroxides, and (d) separating said beads from the liquid phase and drying to achieve the desired porous mixed oxide structure for the beads.

Abstract: The adsorbent consists of a solid phase, completely or partially penetrated by or surface-coated with a hydrophilic molecular polymer netting to which has been bound substituents or cross bridges having chain sequences of the structureX--CH.sub.2 --CH.sub.2 --SO.sub.2 --CH.sub.2 --CH.sub.2 --S--Ywhere X is ether oxygen, a thioether sulfur atom or a nitrogen atom and Y is a substituted or unsubstituted alkyl, aryl or heteroaromatic group. The solid phase consists of particles with a diameter of less than 1 mm and the molecular polymer netting consisting of a cross-linked polyhydroxy polymer such as a polysaccharide, preferably a polygalactane such as agar or agarose or a cross-linked polyamide, e.g. polyacryl amide. Y can be hydroxy alkyl or mercapto alkyl, or phenyl alkyl or phenyl substituted with halogen or nitro groups. The adsorbent is prepared by first converting in a known manner a hydrophilic polymer containing OH and/or CONH.sub.

Abstract: The invention relates to ATP: polynucleotide adenylyltransferase enzyme and a method for its preparation comprising extraction from monocotyledonous plant tissue and chromatographic separation.

Abstract: A method is disclosed for the recovery of biological material from an aqueous solution comprising contacting a water-insoluble, particulate binder with a solution containing biological material to produce a water insoluble biological material/binder composition which may then be recovered. The aqueous solutions may then be recycled.

Abstract: The subject of the application is a process and a reagent for the differentiated determination of the bone and liver isoenzyme of alkaline phosphatase in a sample, whereby this is mixed with a lectin which is able to bind N-acetylglucosamine residues and is incubated. Thereafter, there is carried out a separation of the lectin-bound from the free isoenzyme portions and the alkaline phosphatase activity is determined in one or in both of the separated media.

Abstract: The disclosure is directed to the resolubilization of an insoluble glucose isomerase-amine complex, wherein the amine has the general formula: ##STR1## The insoluble enzyme complex may be resolubilized to produce a stable concentrated and purified glucose isomerase preparation by reaction with a resolubilizing mixture comprising a cation exchange resin.

Abstract: A phosphonamidate or phosphonate analog-ligand having a conformation that substantially corresponds to the conformation of a hydrolytic transition state of an amide or ester ligand is used to produce antibodies of predetermined specificity. The antibodies include an epitope that binds to and thereby stabilizes the tetrahedral carbon atom of the amide or ester hydrolysis transition state of the ligand to hydrolyze the ligand at a predetermined site.

Abstract: This invention relates to a novel process for the recovery of enzyme crystals. The enzymes may be obtained from any enzyme-producing microorganisms such as bacteria, fungi, and yeasts. The invention contemplates supersaturation and/or crystallization to obtain enzymes in the crystalline form, and is particularly effective for the recovery of heat stable alpha-amylase in a crystal form.

Abstract: Solutions containing peroxidase enzymes may be treated to increase their stability and shelf life by reducing their protease level and filtering through a microporous filter to remove any microbial contamination that might secrete additional protease enzymes.

Abstract: Carboxypeptidase B enzyme is isolated by a process wherein homogenized and autolyzed pancreas is subjected to a thermal treatment, and carboxypeptidase B is separated by fractionation with ammonium sulfate. This process produces carboxypeptidase B having satisfactory specific activity while eliminating the use of pancreas cell fluid or an acetonous powder from pancreas as a starting material. The isolated carboxypeptidase B may be immobilized on a polymer gel containing carboxy groups activated with a carbodiimide.

Abstract: The present invention provides a process for the determination of N-carbamoylsarcosine, wherein a sample solution containing N-carbamoylsarcosine is reacted with N-carbamoylsarcosine-amidohydrolase to give sarcosine, which is then determined.The present invention also provides the enzyme N-carbamoylsarcosine-amidohydrolase, a process for obtaining it and a reagent containing it.

Abstract: Substantially pure rubber polymerase and analogues thereof are disclosed. Substantially pure rubber polymerase is isolated and purified from chemically stabilized Hevea brasiliensis latex. The analogues exhibit substantial homology to the Hevea brasiliensis polymerase or function in an enzymatically equivalent manner. Methods for the isolation and purification of a rubber polymerase are set forth. Use of substantially pure rubber polymerase to produce natural rubber or related copolymers is also disclosed.

Abstract: The disclosure is directed to the treatment of a glucose isomerase extract with an amine having the general formula: ##STR1## An insoluble complex containing enzyme activity is formed. The insoluble enzyme complex may be resolubilized to produce a stable concentrated and purified glucose isomerase preparation.

Abstract: A process for the preparation of an enzyme extract containing glucose 6-phosphate dehydrogenase, glucokinase, pyruvate kinase and fructokinase, derived from microorganism cells, by subjecting Zymomonas mobilis bacterium cells to extraction with an extraction medium comprising a partially water-miscible organic solvent; a non-ionic surfactant; and lysozyme; under neutral to alkaline pH conditions to provide an extract containing said enzymes.

Abstract: This invention relates to a process for the production of a purified glucose isomerase enzyme which comprises contacting an enzyme extract containing glucose isomerase and impurities with a first polysulfone membrane not normally permeable to glucose isomerase, in the presence of a salt concentration capable of selectively inducing permeation of glucose isomerase through the membrane, and obtaining a glucose isomerase containing permeate.

Abstract: A process for removing the water from an enzyme-containing aqueous medium is disclosed which comprises spraying the medium onto a heated, fluidized bed of inert substrate particles and recovering a dry enzyme concentrate therefrom.

Abstract: A method and device is disclosed for the release and separation of substances such as parasites or parasite eggs from meat. In the method, meat is agitated with pepsin and filtered through a series of at least two filters, the last of which retains the parasites or parasite eggs. The device includes a reactor which tapers downwardly, and which has in its lower part an outlet valve connected via a conically widening part with a separator device. The separator device includes a cylindrical connecting piece which accepts holder rings for holding one or more filters or sieves in releasable connection.

Abstract: An enzyme contaminated by bacteria during preservation or repeated use thereof is sterilized by immersing the enzyme contaminated mass in a polyvalent alcohol. This procedure does not inactivate the enzyme.

Abstract: The invention is directed to a process for the purification of the dextran-sucrase produced by Leuconostoc mesenteroides bacteria.The process according to the invention consists of adding to the culture medium, that contains the extra-cellular enzyme and dextran, a quantity of a PEG type polyether so that, in the medium appear two non miscible phases, that are thus maintained under stirring in order to obtain a good contact. Thereafter the lower dextran phase is separated from the upper polyether phase in order to provide a dextran-sucrase enriched enzymatic preparation.The purified enzyme can be used in the synthesis processes of the dextrans possessing specific molecular weights for certain applications.

Abstract: The present invention provides an efficient process for purifying the enzyme phenylethanolamine N-methyltransferase suitable for use in radioenzymatic assays of endogenous compounds.

Abstract: The aspartase from a mutant Bacillus subtilis, NRRL B-15536, is produced in relatively high cell yield within a comparatively short time. The enzyme converts fumaric acid to L-aspartic acid stoichiometrically with outstanding selectivity and productivity. The enzyme is stabilized by divalent magnesium ions, 2-mercaptoethanol, and ammonium furmarate, and can be conveniently purified with high recovery.

Abstract: Cyclase, epimerase and a ring expansion enzyme are isolated separately from a cell free extract of a prokaryotic beta-lactam producing organism to provide three separate and stable enzyme reagents for commercial production of cephalosporins from peptide precursors. Isolation is carried out by using ammonium sulfate, gel filtration and ion exchange chromatography. The enzymes may be immobilized on a suitable support and the production of cephalosporins may be carried out continuously.

Abstract: Aminoacylase is isolated from mammal kidneys by comminuting and homogenizing mammal kidney in water, centrifuging to form an aqueous extract, heating the aqueous extract at 60.degree. to 80.degree. C. for 5 to 15 minutes, centrifuging, adding a salt such as ammonium sulfate to the resultant supernatant, centrifuging to separate solids, dissolving the solids in water, dialyzing and recovering active aminoacylase from the dialyzed solution. The process produces aminoacylase with high specific activity and the process requires fewer purification steps. The aminoacylase obtained may be utilized directly without any subsequent purification to produce immonobilized aminoacylase. Immobilization can be carried out by covalent bonding of the aminoacylase to a partially hydrolyzed Akrilex P type acryl amide-N,N'-methylene-bis(arylamide) copolymer. The recovered aminoacylase can be subjected to two chromatographic purification steps to produce a very pure aminoacylase.

Abstract: Glucoamylase and acid fungal protease co-produced by the fermentation of fungal species capable of producing these enzymes are separated by carrying out the fermentation at a pH at which the protease associates itself with the fungal mycelium and removing the mycelium from the fermentation broth. The mycelium is then resuspended in an aqueous medium and the pH adjusted to cause the dissociation of the protease and mycelium with subsequent recovery of the protease from the aqueous medium.

Abstract: A high potency calf rennet substitute having four proteolytic enzyme components is extracted from the stomach mucosa or whole stomach of seals. The substitute closely matches the characteristics of calf rennet by having good milk clotting efficiency over a wide range of pH of 6.0 to 6.8, a high ratio of milk clotting to proteolytic activity, relative inability to inactivate ribonuclease and limited hydrolysis of casein. The substitute results in a desirable aged flavor in cheddar cheese.

Abstract: A method for the identification of microorganisms comprises adding to a sample containing an unknown microorganism an emissive agent such as a radioactive amino acid to produce a mix of emissive products that depends on the metabolic mechanism of the microorganism. After incubation, the reaction is arrested and the emissive products are separated, as by electrophoresis on a gel plate. The plate may then be autoradiographed by exposure to a photographic film to produce on the latter a characteristic band pattern functioning as an identifier for the microorganism. Identification can be effected by comparing the identifier for the unknown with a collection of identifiers for known microorganisms to find a match with one of these known identifiers. The comparison may be carried out by scanning the unknown identifier to produce a signal which is compared with signals representing known identifiers stored in a computer.

Abstract: The enzymes, cyclase, epimerase and a ring expansion enzyme, are isolated separately from a cell-free extract of a prokaryotic beta-lactam producing organism. The isolated enzymes are used to produce unnatural cephalosporins from polypeptide precursors. Isolation is carried out by adding ammonium sulfate to 40% saturation to the cell-free extract to precipitate contaminating proteins, adding ammonium sulfate to 70% saturation to the resultant supernatant to precipitate the enzymes, suspending the precipitated enzymes in a buffer, separating epimerase from the suspension by gel filtration, and separating cyclase and the ring expansion enzyme from each other by ion exchange chromatography.

Abstract: Extracellular protease and amylase coproduced during the fermentation of a microorganism capable of producing them are separated by the addition of polyethyleneglycol and a cationic epihalohydrin/polyamine copolymer or dextran polymer to the fermentation medium and allowing the polymers to phase separate to form a protease rich phase and an amylase rich phase.

Abstract: In the process of crystallizing egg white lysozyme by cooling a salt solution containing egg white lysozyme, a seed crystal is added to the solution, which solution is then maintained at a temperature of about 10.degree. C. to 28.degree. C. for a certain period of time and subsequently cooled, whereby lysozyme can be crystallized to form an agglomerate of many individual needle crystals. The solution containing crystalline lysozyme thus obtained can be filtered within an extremely short period of time, that is, within only a few minutes.