Abstract: Intestinal cell endogenous heparin mediated absorption of cholesterol or fatty acids in mammals is inhibited through the oral administration of heparin, an active heparin subfraction or heparinase. Suppression of cholesterol esterase-mediated absorption in humans can be inhibited by soluble heparin by two mechanisms, i.e. displacement of the enzyme from the intestinal cell membrane and inhibition of enzymatic activity of the displaced enzyme.

Abstract: The invention concerns a new 6-phosphogluconolactonase which has an enzyme activity of at least 90% with respect to the initial activity after about 20 hours at pH 7.0 and 20.degree. C. In addition a process is described for the production of the enzyme from Leuconostoc. The enzyme is preferably suitable for the determination of ATP or reaction partners which can be converted into 6-phosphogluconolactone.

Abstract: This invention provides a method for killing fungal cells without lysing in fermentation processes in order to prepare the fermentation mixture for processing to recover or extract an extracellularly expressed enzyme from the fermentation mixture. A preferred method of this invention comprises adjusting the pH of the fermentation mixture to less than 2.79 using a mineral acid, then adding sufficient acetic acid to the mixture to affect a substantially complete cell kill in mixture. A salt of the acetic acid can be used. The organic acid or salt can be added, then the pH adjusted to the desired level. Other organic acids can be used, in which case the pH of the mixture is adjusted to the pK.sub.a of the selected organic acid before the organic acid is added to the mixture.

Abstract: Disclosed is a method for the extraction of lipase from mammalian tissue with which it is associated which involves contacting the tissue with an alkaline, aqueous medium having a pH of from greater than 7.0 up to a level at which the alkalinity will deactivate the lipase.

Abstract: The present invention provides methods for detecting and quantitating BCR-ABL gene products and other abnormal ABL gene products of Ph.sup.1 -positive leukemic cells. The invention further provides methods for determining the relative number of leukemic cells compared with normal ABL cells to assess the tumor burden of a patient. In another aspect, the methods of the present invention can be used to determine a specific phase of leukemia, particularly chronic-phase CML.

Abstract: A purified low molecular weight endoglucanase II from Acidothermus cellulolyticus (ATCC 43068) is disclosed. The endoglucanase is water soluble, possesses both C.sub.1, and C.sub.x types of enzyme activity, a high degree of stability toward heat, and exhibits optimum temperature activity at about 81.degree. C. at pH's from about 2 to about 9, and at a inactivation temperature of about 100.degree. C. at pH's from about 2 to about 9.

Abstract: A method for recovering a purified 52,000 dalton fraction of human pancreatic cholesterol esterase is disclosed. The 52,000 dalton fraction of human pancreatic cholesterol esterase is purified by passing a solution having a pH of about 8.2 which contains two or more fractions of human pancreatic cholesterol esterase including the 52,000 dalton fraction over DEAE-cellulose to produce an effluent containing said 52,000 dalton fraction. Since the 52,000 dalton fraction of the esterase does not bind to the DEAE-cellulose whereas the other esterase fractions do bind to the DEAE-cellulose, the 52,000 dalton fraction is then collected in the effluent. The method can be further carried out by passing the purified 52,000 dalton fraction over both hydroxyapatite and heparin-Sepharose to obtain a homogeneous solution of the 52,000 dalton fraction.

Abstract: Disclosed is a method of producing an extracellular product from an aerobic microorganism. The microorganism is grown in an aqueous medium on one side of an oxygen-permeable surface while the opposite side of the surface is contacted with oxygen. The extracellular product is then harvested from the aqueous medium. The method is particularly useful in producing lignin peroxidase from the white rot fungus Phanerochaete chrysosporium.

Abstract: Recombinant protein purification vectors and methods for their use are disclosed. The vectors contain a DNA sequence coding for a gelatin binding region of fibronectin. The vectors express a foreign DNA sequence of interest fused to the fibronectin portion. Secretion signals on the fused product assist the product in being secreted from a production cell. The product can then be purified on a gelatin containing affinity column and digested with a protease such as trypsin to cleave the desired protein from the gelatin binding region. The vectors can also be designed to code for factor XIIIa cross liking sites and to have a chemically reactive cysteine residue.

Abstract: Novel methods and compositions are provided for enhanced yield of heterologous proteins in fungi. The method and compositions involve employing fusion sequences involving a sequence encoding a heterologous product produced in relatively large amount as a stable polypeptide in the host fused to a second sequence in open reading frame with the prior sequence coding for a different heterologous polypeptide, where the two polypeptides are joined by a selectively cleavable linkage. In particular, a sequence coding for superoxide dismutase is joined to another polypeptide of interest at either terminus of the superoxide dismutase in a yeast expression vector under transcriptional control of an active promoter and the vector introduced into a yeast host and the host grown. High yields of the fusion product are obtained in this manner, where the fusion product can be selectively cleaved so as to produce both the superoxide dismutase and the other polypeptide in high yield.The S.

Abstract: The preparation of a pepsin extract from the stomach of pigs, sheep or cattle, which is suitable for the bating of hides, is presented. The process involves macerating the stomach mucous, sedimenting the material, reserving the middle layer for pharmaceutical preparation, combining the other two layers, drying them and adding salt for preservation. The resulting pepsin containing material is particularly suitable for acid enzymic bating of chrome tan hides after the pickle process with the main purpose of obtaining a higher surface yield and a softer leather.

Abstract: A process for the separation of streptokinase from contaminating proteins in a streptokinase-containing mixture, which comprises treating the mixture with a reducing agent to reduce disulphide bridges in the contaminating proteins to free thiol groups, contacting the mixture with a reagent R-X wherein R is a group capable of reacting with a free thiol group and X is a group R.sup.1 capable of reacting with a free thiol group or is a thiol-containing matrix, and thereafter separating the resulting chemically modified contaminating proteins from the mixture to provide streptokinase in a form substantially free of contaminating proteins.

Abstract: A method for preparing an aqueous solution enriched in xylanase from an aqueous mixture containing cellulase proteins and xylanase is disclosed. The method involves adding an amount of a low molecular weight alcohol to an aqueous mixture containing cellulase proteins and an organic salt so as to precipitate out the cellulase proteins other than EG III from the mixture. The method then involves adding an inorganic salt to the supernate produced in the previous step so as to form a second precipitate and a second supernate and then finally collecting the second precipitate from the second supernate to obtain a precipitate enriched in xylanase.

Abstract: A denatured human O.sup.6 -guanine alkyltransferase is disclosed. The enzyme is prepared by a process which involves denaturing the enzyme, contacting the denatured enzyme with a monoclonal antibody specific for the denatured enzyme on a substrate to which the monoclonal antibody is bound to, so that the denatured enzyme and the monoclonal antibody form an immunocomplex and then, eluting the denatured enzyme from the substrate-bound monoclonal antibody so that the denatured human O.sup.6 -guanine alkyltransferase is obtained. The enzyme can be used to develop probes for the Mer.sup.- phenotype and these probes in turn are contemplated for use in identifying drug resistant tumors in patients.

Abstract: The invention relates to NAD(P)H-dependent nitrate reductase from yeasts, to a process for the preparation thereof and to the use thereof in a reagent for determining nitrate. The nitrate reductase is characterized by a molecular weight of about 350 000 D and can be obtained by yeast cells which have been cultivated in a completely synthetic nutrient medium with nitrate as the sole nitrogen source and which contain nitrate reductase being disrupted in phosphate buffer, the crude extract being chromatographed on an anion exchanger, the fractions containing nitrate reductase being mixed with protein, concentrated by ultrafiltration and dried by fluidized bed granulation. The reagent for determining nitrate contains, besides the nitrate reductase prepared in this way, also NAD(P)H and a color reagent for determining nitrite.

Abstract: A process for the manufacture of purified thromboplastin used in the manufacture of an ultra-pure, clear thrombin solution is described.

Abstract: The present invention provides the identification and characterization of two components of a recombinant preparation of DNase. These components are the purified deamidated and non-deamidated human DNases. Taught herein are the separation of these components and the use of the non-deamidated species as a pharmaceutical per se, and in particular in compositions wherein the species is disclosed within a plastic vial, for use in administering to patients suffering from pulmonary distress.

Abstract: Affinity chromatography matrices are disclosed which are useful in purifying interleukin-1.beta. converting enzyme (ICE) from crude or partially purified cell lysate preparations. The chromatographic matrices comprise a specific ICE inhibitor of Formula I which is attached to an affinity column support by means of a bifunctional spacer.

Abstract: This disclosure relates to a novel class of serine proteases isolated from a culture medium of fungus Tritirachium album. The serine proteases disclosed have a high degree of stability in detergent formulations.In addition, this disclosure relates to a process for producing such serine proteases using recombinant techniques.

Abstract: A purified low molecular weight cellulase endoglucanase I having a molecular weight of between about 57,420 to about 74,580 daltons from Acidothermus cellulolyticus (ATCC 43068). The cellulase is water soluble, possesses both C.sub.1 and C.sub.x types of enzyme activity, a high degree of stability toward heat, and exhibits optimum temperature activity at about 83.degree. C. at pH's from about 2 to about 9, and in inactivation temperature of about 110.degree. C. at pH's from about 2 to about 9.

Abstract: The invention proposes a process for purifying a highly glycosylated protein from a crude preparation which comprises the action (i) of adding to said preparation a divalent metal ion in a sufficient amount in order to form a mixture which precipitates and (ii) after precipitation, of harvesting said protein from the mixture supernatant.

Abstract: A method of assay for isotopically exchangeable analytes is disclosed. Analytes are labeled by enzymatic exchange of a hydrogen atom of the analyte and a deuterium or tritium atom. Preferably, analytes are labeled by reaction with an oxidant, a reducing agent which contains a deuterium or tritium atom, and an enzyme capable of catalyzing the reversible exchange of a hydrogen atom between the analyte, the oxidant, and the reducing agent. Kits for conveniently performing the assay methods are also disclosed.

Abstract: Butyrylcholinesterase is produced in a purity of at least 90% by subjecting plasma fraction IV-4 alone or in admixture with fraction IV-1 to both anion exchange chromatography and affinity chromatography.

Abstract: Invertase is produced by a process wherein, first the yeast cells are disrupted to produce a disrupted cell suspension, second the disrupted cell suspension is adjusted to an acidic pH, third denatured undersired proteins are removed with the cell detritus and lastly the invertase is isolated. The improvement of the invention to the state of the art is that it comprises subjecting the disrupted cell suspension prior to removing the undesired proteins and cell detritus, to a pH of less than 4.5 and to a heat treatment in a continuous thermal denaturation system at a temperature in a range of from 44.degree. C. to about 51.degree. C.

Abstract: A culture of Pseudomonas XA cells containing indolyl-3-alkane alpha-hydroxylase(INDH) is produced by aerobic batch fermentation in a culture medium. The culture medium is in a vessel having a means for adjusting agitation rate. During culturing, the cells go through a logarithmic phase and a stationary phase. Oxygen concentration is measured in the culture medium at the beginning of the process to determine a first oxygen concentration. Agitation rate is adjusted to produce a second oxygen concentration in the culture medium of about two to about seven percent of the first oxygen concentration for a time period of about two to about seven hours which ends at the beginning of the stationary phase. An INDH having three subunits of different molecular weight is isolated from the cells. The INDH is stable, free from endotoxin-mediated side effects and has sufficient specific activity to be useful for depletion of tryptophan from aqueous solution. During use, the INDH may be in immobilized form.

Abstract: A methionine N.sup..alpha. -acetyltransferase enzyme has been identified in yeast. The existence of the enzyme defines a gene for methionine N.sup..alpha. -acetyltransferase which may be cloned and altered. Cells expressing the normal or variant forms of this enzyme are valuable tools for determining the amino acid sequence of uncharacterized proteins. Such cells are also valuable tools for expressing a recombinant protein lacking an acetyl group at its .alpha.-amino group.

Abstract: A Triqonopsis variabilis D-amino acid oxidase in substantially pure form and active against cephalosporin C is disclosed. This D-amino acid oxidase is isolated from Trigonopsis variabilis by a method which is performed in three steps, namely:(a) acidifying and heating a crude cell extract of Trigonopsis variabilis to obtain a precipitate and supernatant fraction;(b) treating said supernatant fraction obtained in step (a) with sufficient ammonium sulfate to obtain a second precipitate, said second precipitate containing the D-amino acid oxidase of claim 1; and(c) resuspending the precipitate obtained in step (b) and collecting the D-amino acid oxidase by isoelectric precipitation.

Abstract: A process for the purification of plasminogen activator inhibitor 2 (PAI-2) including the steps of preincubating a PAI-2-containing solution with a compound which cleaves disulfide linkages, mixing the solution with a water-soluble acridine or quinoline base to precipitate impurities in the solution, and separating the precipitated impurities to obtain PAI-2 in purified form.

Abstract: A proteolytic factor is capable of cleaving .beta.-protein precursor at a site near the .beta.-protein N-terminus. Also, a method for treating Alzheimer's disease in a patient includes steps of reducing .beta.-protein precursor proteolysis outside the .beta.-protein domain at a site near the .beta.-protein N-terminus. Also, a method for purifying an enzyme from a sample includes steps of incubating the sample with a labelled substrate of the enzyme or with a labelled fragment of a substrate to which the enzyme binds, treating the sample with a crosslinking agent to crosslink any enzyme-substrate complexes in the sample, and recovering labelled complexes. Also, a method for diagnosis in a subject of a disease characterized by accumulation of amyloid includes determining the level, in a sample of tissue or body fluid from the subject, of an AD proteolytic factor.

Abstract: A biochemical reagent comprises an oligosaccharide, preferably one which has been liberated from an immunogenic glycoprotein or proteoglycan, which is immobilized on a carrier via an intermediate spacer molecule such as a lipid. The lipid molecule should preferably have at least two long lipid tails so that the oligosaccharide is held in spaced relationship to the carrier where is exhibits antibody-binding ability which is almost indistinguishable from that of the original glycoprotein or proteoglycan. The reagent has its application in biochemical testing of oligosaccharides and systems which bind to them.

Abstract: In a process for growing enzyme crystals, small crystals are continuously removed from a crystallizer, dissolved and returned to the crystallizer to maintain a supersaturated state. The method permits the growing of large crystalline enzymes of uniform size of about 0.5 to 1 mm. Solid materials can be coated with crystalline enzymes by placing a solid material in the crystallizer such that crystals deposit on the solid material. The process is preferably used to produce crystalline glucose isomerase.

Abstract: A method for obtaining DNA expressing histidine rich protein of various types of Plasmodia is disclosed. The method involves hybridization with the comparable DNA of P. lophurae. The method of particularly well suited for obtaining P. falciparum DNA, whether it is associated with know or knobless phenotype. Additionally, the invention disclosed a safe method for diagnosing P. falciparum infection.

Abstract: Biocatalytic oxidative processes wherein oxidizable substrates are reacted with peroxides in the presence of a peroxidase such as soybean peroxidase or another legume peroxidase; an oxidative coupling reaction for producing phenolic resins; a method for the purification of peroxidase enzyme-containing extracts generally also are described.

Abstract: A novel urease having an optimal pH for activity in the acidic region is produced by a microorganism belonging to the genus Lactobacillus or Streptococcus. The urease is superior to the conventional urease in pH stability, temperature stability and alcohol stability.

Abstract: An aqueous two-phase protein partitioning system is disclosed which employs polyvinylpyrrolidone as the upper phase and maltodextrin as the lower phase and provides a low-cost system for protein partitioning. The system can also be employed with the amion derivatives of chlorotriazine dyes, which bind in a noncovalent manner with the PVP and serve as a ligand for the proteins to be separated.

Abstract: 2-O-.alpha.-D-Glucopyranosyl-L-ascrobic acid is crystallizable in its supersaturated solution. Crystalline 2-O-.alpha.-D-glucopyranosyl-L-ascorbic acid is substantially nonhygroscopic, free flowing, free of deliquescence, consolidation and direct reducing activity, but is readily soluble in water. Because of these, crystalline 2-O-.alpha.-D-glucopyranosyl-L-ascorbic acid is handleable with an ease, and superiorly high in stability and physiological activities. Thus, crystalline 2-O-.alpha.-D-glucopyranosyl-L-acorbic acid is favorably useful in vitamin C-enriching agents, foodstuffs, pharmaceuticals and cosmetics.

Abstract: A novel lipase from a newly-discovered strain of Pseudomonas alcaligenes microorganism having (i) an optimum pH for activity of about 10.+-.0.5; (ii) an optimum temperature for activity of about 45.degree. to 55.degree. C.; (iii) an optimum pH for stability of about 7.0.+-.0.5; (iv) a molecular weight as measured by gel permeation chromotagraphy of about 8.8.times.10.sup.4 ; and (v) chemical stability for at least 30 days in the presence of the surfactants. Also claimed is a biologically pure culture of the microorganism, and a method for the production of the lipase.

Abstract: A thermoduric and aciduric pullulanase enzyme, and a process for its production from a microorganism designated as Bacillus naganoensis. The pullulanase is useful for the production of dextrose and high-maltose syrups from starch hydrolyzates.

Abstract: Method for producing a proteolytic complex formed after cultivation of a strain of Streptomyces fradiae while maintaining the pH of the culture medium between 7 and 7.5, and extracting said complex by filtration in the presence of a clarifying agent and ultrafiltration on a specific membrane and finally atomization; the invention also relates to the new proteolytic complex thus obtained and its applications in zootechny due to its stimulating effect on the activity of the pancreas.

Abstract: The instant invention is a process for the recovery from an aqueous solution of water soluble biopolymers, in particular polysaccharides, gums, agar, agarose and starches by the addition of polyoxide solution to the biopolymer containing solution. The biopolymer is then separated and collected.

Abstract: Crystalline subtilisin is produced by adding a halide salt, such as sodium chloride or calcium chloride, to a concentrated subtilisin solution (at least about 40 g/1). This process does not produce amorphous subtilisin even at high salt concentrations in the solution. Optionally, subtilisin seed crystals also may be added to the concentrate to speed up the crystallization process.

Abstract: A process is described for the purification of plasminogen activator (PA), wherein a solution containing such as plasminogen activator is brought into contact with a carrier-bound polysulfate of a saccharide or sulfated sugar, the liquid is removed, and the PA bound by this material is eluted.

Abstract: A method for purifying an aqueous peroxidase enzyme containing solution and peroxidase solution purified thereby are disclosed; the method includes the steps of:providing an aqueous solution containing a peroxidase enzyme;adding a water-immiscible or partially water-miscible organic solvent to said aqueous solution to form a mixture having an organic phase and an aqueous phase;agitating said mixture to extract impurities from said aqueous phase into said organic phase;separating said aqueous phase from said organic phase; anddiscarding said organic phase.

Abstract: A novel acetylpolyamine amidohydrolase is described. The enzyme specifically hydrolyzes the amino bond in acetylputrescine, acetylcadaverine, acetylspermidine and acetylspermine with strong substrate affinity. The enzyme is preferably produced by culturing a microorganism belonging to the genus Mycoplana, and is used in the quantitative determination of polyamine contained in a living body sample, which is useful for cancer diagnosis.

Abstract: The present invention provides a restriction endonuclease which recognizes palindromic sequences ##STR1## where C* is methylated, and cleaves these sequences at the position indicated by the arrows. This endonuclease is preferably from a microorganism of the genus Kluyvera. The present invention also provides a process for obtaining this new restriction endonuclease and a method for using the endonuclease.

Abstract: The present invention provides for the use in the production of cellulolytic and xylanolytic enzymes, particularly xylanase and cellulase, of the microorganism Thermoascus aurantiacus in a culture medium containing at least one of a cellulose or hemicellulose substrate whereby to produce thermostable enzymes, particularly cellulase and xylanase.

Abstract: Enzymes isolated from krill of the order Euphausiaceae are used to remove biological contaminants. Preferably, a mixture of enzymes including exo-and endopeptidase is isolated. The enzymes can be used in laundering or to clean or debride living tissue. Isolation may be carried out by homogenizing krill and extracting the enzymes with an aqueous medium. The enzymes may be further purified by gel chromatography. After lipids have been removed, the enzymes can be lyophilized for long time storage.

Abstract: Gas containing granules of a resiliently compressible material having a density variable with pressure are used as a solid phase support in liquid phase reactions such in immunological reactions. Varying pressure causes the density of the granules to change and is used for mixing and separating the granules in liquid phase. In an immunological reaction, the granules having an attached antibody are combined with a solution containing an antigen, then varying pressure is applied to the solution to mix the granules in the solution to fix the antigen to the antibody and then pressure on the solution is varied to force the granules to the top or bottom of the solution whereby the solution can be separated from the granules.

Abstract: Antifoaming agents are commonly used during culturing of enzyme-producing microorganisms. These antifoams often persist through enzyme processing, slowing filtrations, clogging filtration membranes and adversely affecting the quality of the final product. The present invention describes a method for removing antifoams, and often carbohydrates and pigments, from enzyme systems using mineral clay, the preferred clay being bentonite.

Abstract: The invention relates to a process for producing L(-)-tetrahydrofolic acid which comprises allowing dihydrofolate reductase to act upon dihydrofolic acid in the presence of (1) NADP or NADPH, and (2) glucose and glucose dehydrogenase. L(-)-tetrahydrofolic acid is useful as the intermediate for L(-)-leucovarin.