Abstract: A process for the recovery of enzymes from an aqueous system having at least two aqueous phases wherein the partition coefficient with respect to an enzyme may be improved through the addition of organo-compatible, compounds having anionic functionality, such as polyacrylic acid, polyvinyl sulfonate and sodium dodecylsulfate.

Abstract: A method is provided for increasing the solubility of an enzyme in an aqueous solution comprising providing a water-soluble anionic surfactant to the solution and further providing the solution with a pH above that which result in the formation of an enzyme/surfactant precipitate. The application is further directed to an aqueous solution containing an anionic surfactant and having an enzyme concentration contained therein which is higher than that possible in the absence of the surfactant.

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: A process for the extraction and activation of microbially produced bovine prochymosin is disclosed.

Abstract: Enzymes extracted from a prokaryotic .beta.-lactam producing microorganism are immobilized on a support for producing desacetoxycephalosporin and analogs thereof from L-.alpha.-aminoadipyl-L-cysteinyl-D-valine and analogs thereof. The enzymes are an epimerase, a cyclase and a ring expansion enzyme extracted preferably from S. clavuligerus, S. cattleya or S. lipmanii. The support is preferably a diethylaminoethyl ion exchange resin.

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: This invention relates to three enzymes, their isolation from the viscera of bivalves, e.g. the surf clam or cherrystone clam, their characterization and uses. The first two are carboxyl proteinases having molecular weights of about 77,200 and about 36,700 and display activity similar to mammalian D-cathepins. The third is a thiol proteinase having a molecular weight of about 17,400 and displays activity similar to mammalian B-cathepins. In addition to attaching various substrates, the enzymes coagulate cheese milk and tenderize meat.

Abstract: This invention relates to a novel process for the recovery of enzymes obtained from enzyme-producing microorganisms, and to the liquid enzyme product recovered by this process. Typically, the enzyme-containing filtrate from a fermentation of an enzyme-secreting microorganism is concentrated and a precipitation agent such as a salt or an organic solvent is added to the concentrate, thereby forming a cake. Then, a polyol solvent is circulated through the cake to solubilize the enzyme or enzyme complex from the cake and provide a liquid enzyme product. Particularly effective is propylene glycol as the polyol solvent. The liquid enzyme product may be shipped as is or subjected to further treatment to remove the solvent and create an essentially solvent-free enzyme product. The process is especially effective for the recovery of alkaline protease or alpa amylase.

Abstract: A method of specimen treatment preparatory to conducting an immunoassay is disclosed whereby a microbial protein is solubilized by a detergent at elevated temperatures and in the presence of an alkali or alkaline earth metal ion. At elevated temperatures, the detergent is soluble. However, at lower temperatures, the presence of the metal ion renders the detergent insoluble so that it is prevented from interacting in the immunoassay procedure. A specific application is in the solubilization of the principal outer membrane protein of Chlamydia trachomatis.

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: Meat is tenderized by adding thereto a proteolytic enzyme obtained by culing the microorganism, Trichoderma reesei strain MCG 80. The enzyme is an aspartic acid protease with proteolytic properties similar to the animal protease, Cathepsin D. The enzyme acts selectively upon the myofibrillar proteins of meat producing a desirable uniform texture. Culturing of the microorganism in a medium containing glucose and lactose results in high enzyme yield.

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: An improved method for the purification of ribulose, 1,5-bisphosphate carboxylase (RuBisCO) comprises comminuting and homogenizing a plant material, such as leaves, in an aqueous solution. After fractionation to release the RuBisCO, sufficient polyethylene glycol (PEG) is added to cause crystallization of the RuBisCO. It has been found that treatment of the resulting PEG supernatant first by acidification to remove other proteins, and then by addition of a strong base to remove phosphorylated sugars, allows the recycling of the PEG in the process. Moreover, it is found that the phosphorylated sugars are a valuable by-product, suitable for example as a carbon source for the culture of microorganisms.

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: A new alcohol oxidase is isolated from Pichia-type microorganisms in soluble or crystalline form. The crystalline novel enzyme is isolated by preparing an aqueous fluid containing cells of a Pichia-type microorganism, homogenizing the fluid and separating solids therefrom to produce an alcohol oxidase solution, adjusting the solution to have an ionic strength in the range of 0.05 to 0.01 in ionic strength to form a recovery range solution thereby causing the crystalline alcohol oxidase to form. The new enzyme is used to determine alcohol concentrations in fluid samples in which conditions are compatible with the enzyme's activity.

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: Compositions containing proteolytic enzymes having no detectable odor at a concentration of less than about 0.002 Anson units per gram of distilled water, and selected perfume materials for improved odor. Heavy-duty liquid detergents are preferred.

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.

Abstract: This invention relates to a novel process for the separation, purification and recovery of dextranase from impure solutions such as fermentation broth by precipitating a complex of dextranase with tannic acid and thereafter separating the purified dextranase from the tannic acid.

Abstract: Combining active alcohol oxidase with sufficient amount of an azide compound has been found to form a red absorbing combination. Formation of the red absorbing combination enables determining the presence of active alcohol oxidase by adding the azide compound to a preparation and observing the resultant color. Additionally, alcohol oxidase may be purified by adding the azide compound to a preparation containing active alcohol oxidase in an amount effective to produce a red absorbing complex and separating the red absorbing complex from the preparation.

Abstract: For the determination of glycerin by oxidation with oxygen in the presence of glycerinoxidase and measurement of the oxygen consumption or of the H.sub.2 O.sub.2 formation, a glycerinoxidase from Aspergillus spec. DSM 1729 is used. A reagent suitable for this method consists of glycerinoxidase from Aspergillus spec. DSM 1729 and a system for the determination of H.sub.2 O.sub.2 and contains additionally, if desired, an agent for the saponification of esterified glycerin.

Abstract: Novel fibrinolytic factors obtained from the salivary glands of Haementeria ghilianii. Proteins having molecular weights under about 100,000 are isolated from the salivary glands of H. ghilianii. The proteins show cathodic mobility in electrophoresis and uninhibited peptidase activity with fibrinogen in plasma.

Abstract: A debranching enzyme (pullulanese) useful in the preparation of a low calorie beer may be obtained from rice by extraction of the rice with an aqueous buffer system having a pH of about 6. A preferred buffer system is 0.1 M potassium phosphate-0.2 M-sodium chloride. Extraction is preferably carried out at a temperature of about 50.degree. C. for about 3 hours. When malted rice is used as the enzyme source a particularly useful mixture of the debranching enzyme and alpha 1,4 carbohydrases is obtained.

Abstract: Low calorie beer is prepared by introducing into the brewing process a debranching enzyme (pullulanase) obtained from rice, a traditional brewing material. The debranching enzyme reduces the real extract of the beer by cleaving alpha 1,6 linkages of unfermentable limit dextrins to form alpha 1,4 dextrins which can be converted by alpha 1,4 carbohydrases to sugars that can be fermented by brewer's yeast. The enzyme may be introduced into the brewing process by adding rice or the enzyme extracted from rice to the mash or to the wort before or during fermentation. The debranching enzyme may be obtained from polished dry milled rice by extraction with an aqueous buffer solution. When malted rice is used as the enzyme source a particularly useful mixture of the debranching enzyme and alpha 1,4 carbohydrases is obtained.

Abstract: Described herein is a process for isolating the enzymatic protein ribulose 1,5-diphosphate carboxylase from the green leaves of plants. In the process, which is particularly suited to obtaining the protein from tobacco, the leaves are ground or otherwise pulverized in the presence of a reducing agent. A liquid portion containing the desired protein is separated from the pulp and its pH adjusted to within the range of from about pH 6.0 to 5.3 and then cooled to cause the crystallization of the ribulose 1,5-diphosphate carboxylase. After separation of the crystalline material, the supernatant is acidified to yield lower molecular weight proteins.

Abstract: A process for isolating superoxide dismutase from red blood cells comprising the steps of heating hemolyzed red blood cells in the presence of at least one monovalent inorganic neutral salt and at least one transition metal salt; eliminating precipitate from the hemolyzed red blood cells by filtration or centrifugation to obtain a solution; adding further at least one transition metal salt at least once to the solution under application of heat thereto; and eliminating precipitate from the solution.

Abstract: Various enzymes, in particular Cu,Zn-superoxide dismutase (SOD), catalase and carbonic acid anhydrase, are recovered from blood by admixing wholly or partly isolated blood cells with ethanol or a homologous alcohol until a concentration of 10 to 70% by volume and allowing them to stand for hemolysis of the blood cells and denaturation of the hemoglobin, and then adding water up to the double volume or more, and removing the precipitate of cell residuals, hemoglobin and other denatured proteins from the suspension. Then the desired enzymes are isolated from the solution obtained.In particular, SOD and catalase can both be isolated by chromatography of the solution at a pH of 4.7 to 5.5 on a cation exchange resin of the same polarity as SOD in the pH range used and elution of the resin with a buffer solution which has a pH in the range 4.7 to 7.5 and an ionic strength in the range 0.01 to 1.0 M, SOD being eluted at the lowest pH and/or the lowest ionic strength.

Abstract: Fraction I protein from plant leaves is purified and subsequently crystallized. The crystallization methods disclosed herein unexpectedly produce crystallization in all crop leaves examined, although for some species modification by salt addition is required to achieve crystallization and to prevent formation of substantial percentages of amorphous protein precipitates. It has been found that a fraction I protein solution, when mixed with a precipitant solution having a pH generally within the range of 4.8-7.2, in an amount and at a pH sufficient to provide a mixed solution (protein solution mixed with precipitant solution) having a final pH in the range of 6.6-7.0, causes crystallization of fraction I protein from plant leaves, provided that the precipitant solution is at a pH lower than the pH of the protein solution. Optimum results have been obtained when the pH of the precipitant solution is in the range of 5.0 to 6.0 and the protein solution in the range of 7.0 to 7.5.

Abstract: Improved steps and methods for rapid, efficient and accurate assay of the CPK isozyme MB (cardiac fraction) and other determinations using chromatographic column technique. The improvements are especially adapted for carrying out rapid diagnostic tests in clinics and hospitals. Also contemplated improved apparatus and methods and uses of same adapted for the test method, such apparatus having novel features which can be packaged and merchandised in kit form. Other special features include use of electrophoresis either with or without the conventional chromatographic column analyses and using a selected colored (preferred-green) marker dye. Another special feature is that the used columns of the improved design can be conveniently recycled using the same solutions.

Abstract: A single, simple, uniform protocol for the rapid large scale purification and crystallization of ribulose 1,5-bisphosphate carboxylase (RuBisCO) to a greater than 90% purity from a wide variety of plant species, involving the steps of grinding and homogenizing plant material, such as leaves, with a suitable, acqueous buffer solution. The solution is filtered and the residue discarded. While maintaining temperature and pH control, sufficient quantities of polyethylene glycol (PEG) are added while stirring to bring the final concentration of PEG in the range of 8 to 15 (weight/volume) percent. To enhance crystal formation, magnesium chloride can be added to the solution. The precipitated material is discarded and the pure RuBisCO crystals which separate out of the remaining supernatant are collected, washed, dried and stored.

Abstract: Double-stranded RNA synthesized using native human DNA as template is found to be an excellent interferon inducer with low toxicity in spite of homologous native to host cell. It is produced by reacting ATP, GTP, CTP and UTP with one another in the presence of a native human DNA as template by the catalytic action of an active RNA polymerase to form RNA and subjecting the resulting RNA to annealing by heating it at a temperature of 70.degree. to 100.degree. C. and gradually cooling to room temperature or below to form double-stranded regions between their molecules.

Abstract: Myeloperoxidase can be separated and recovered from a supernatant liquid of an aqueous dispersion of a disintegration product of human myelogenous leukocytes, which dispersion has been admixed with at least one member selected from the group consisting of manganese salts and protamine sulfate. The recovered myeloperoxidase in combination with an alkali metal halide can be used as a pharmaceutical composition effective against microorganisms deficient or diminished in catalase synthesizing activity.

Abstract: A method to inactivate the proteolytic enzyme(s) contained in commercial heat stable bacterial alpha-amylase under conditions which retain full alpha-amylase activity. Use of thus purified alpha-amylase enzyme plus surfactants that are approved for use in bread to inhibit firming and improve the keeping quality of bread and other bakery products.

Abstract: A process is disclosed for isolating an enzyme mixture useful in the treatment of devitalized tissue from crude bromelain. The process comprises the steps of: (1) suspending the crude bromelain in a weakly basic buffer, preferably sodium borate buffer, to selectively dissolve the active enzyme mixture; (2) separating the undissolved solids from the solution; and (3) removing small molecules having a molecular weight of 10,000 or less from the solution.

Abstract: An enzyme preparation for tenderizing meat products is produced from endocrine-enzyme raw stock. The testes of slaughtered cattle are comminuted and mixed with acidulated water to obtain a mixture having the pH value within the range 4.5 to 4.7, followed by isolation of an aqueous solution from said mixture, the solution containing hyaluronidase enzyme. An albumin-containing substance is then added, the ratio of albumin-containing substance to the testes being between 0.04-1.6:10-15, to obtain a suspension containing the end product. The suspension is concentrated by evaporation in vacuo at a maximum temperature of 25.degree. C. to form a concentrate of the end product, which is then dried.

Abstract: A process for the recovery of an intracellular enzyme from an aerobic soil microorganism is disclosed. The recovery method is carried out by(a) forming an aqueous suspension of microbial cells containing the desired intracellular enzyme,(b) disrupting the microbial cells in the suspension to release the enzyme from the cells, and(c) before, during, or after step (b) and prior to removal of disrupted microbial cells and other cellular components, introducing a water-miscible organic solvent into the suspension to form a mixture of the organic solvent and the enzyme-containing suspension.The desired enzyme is retained in the liquid phase of the mixture formed in step (c) while undesired cellular components such as other microbial cell proteins precipitate therefrom.

Abstract: Described herein is a process for isolating the enzymatic protein ribulose 1,5-diphosphate carboxylase from the green leaves of plants. In the process, which is particularly suited to obtaining the protein from tobacco, the leaves are ground or otherwise pulverized in the presence of a reducing agent, followed by heating the resulting pulp to about 50.degree. C. A liquid portion containing the desired protein is separated from the pulp and then cooled to cause the crystallization of the ribulose 1,5-diphosphate carboxylase. After separation of the crystalline material, the supernatant is acidified to yield lower molecular weight proteins.

Abstract: Increased yields of the cellulolytic enzymes from Thielavia terrestris are effected by novel separatory methods for the produced enzymes as well as the enhancement of the production of one of the enzymes, namely .beta.-glucosidase, by the addition of glycerol to the media. Further, since the .beta.-glucosidase is responsible for glucose production from cellulose, a method is provided wherein the separated .beta.-glucosidase can be employed as, for example, on a fixed support in a position to receive concentrated streams of partially converted cellulosic materials, thereby leading to enhanced glucose production.

Abstract: A process for purifying glucose isomerase comprises the steps of acid treatment and salt fractionation. An enzyme solution is treated with an acid, such as acetic acid, to a pH from about 3.5 to about 5.0. The proteinaceous solids are collected and extracted with a buffer, such as imidazole, whose solution has a pH of about 6 to about 8. The solution is then collected and a salt, such as ammonium sulfate, is dissolved therein from about 40% to about 50% of its saturation point. The proteinaceous solids which form are removed and additional ammonium sulfate is dissolved to attain from about 41% to about 60% of its saturation point, followed by collection of the solids containing purified enzyme. A composition which preserves enzyme activity upon storage of glucose isomerase and which imparts resistance to thermal deactivation of said enzyme comprises an aqueous solution of glycerol, a buffer whose solution is at a pH of about 6 to about 8, divalent cobalt ions and magnesium ions.

Abstract: Protease is recovered by reacting a first amino acid whose amino group is protected with a protective group with a second amino acid whose carboxyl group is protected with a protective group, in an aqueous medium in the presence of a protease to result a peptide synthesis to deposit the addition compound of a dipeptide and said second C-terminal protected amino acid; dissolving said addition compound by adding an organic solvent to a precipitate separated from the reaction mixture and isolating the protease from an organic solvent suspension or dissolving said addition compound by adding a water immiscible organic solvent and separating the organic solvent phase by a liquid-liquid separation.The addition compound can be dissolved in an organic solvent after separating the precipitate from the reaction mixture.The addition compound can be also dissolved by adding a water-immiscible organic solvent to the reaction mixture and the organic solvent phase can be separated from the aqueous phase.

Abstract: Protease is recovered by reacting a first amino acid whose amino group is protected with a protective group with a second amino acid whose carboxyl group is protected with a protective group, in an aqueous medium in the presence of a protease to result a peptide synthesis to deposit the addition compound of a dipeptide and said second C-terminal protected amino acid; dissolving said addition compound into the aqueous medium by adding a polar organic solvent which is miscible with water and separating an insoluble material from the resulting suspension by a solid-liquid separation to isolate the protease.

Abstract: A fungal enzyme preparation has specific bilirubin degrading activity and, in a preferred embodiment, generates hydrogen peroxide. Extraction methods for the enzyme preparation are disclosed. Assay compositions, elements and methods using the aforementioned enzyme preparation are also disclosed.