Abstract: A method of improving a ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo) to have a higher protein score is disclosed. The method includes the steps of: making a modified RbcL of the RuBisCo, by, on an RbcL unit of the RuBisCo, either substituting Met for Leu, Phe, Val, or Ile or combinations thereof; substituting Lys for Arg, Thr, or His or combinations thereof; or both of these substitutions. The modified RbcL consequently modifies the RuBisCo and is added to a biomass host where it is stable for homologous recombination. Plastid and nucleus integration was observed. Example RbcL sequences are disclosed with the desirable substitutions. The improved RuBisCo can be used as an improved proteinaceous food source for humans and animals.

Abstract: Genes encoding Class II EPSPS enzymes are disclosed. The genes are useful in producing transformed bacteria and plants which are tolerant to glyphosate herbicide. Class II EPSPS genes share little homology with known, Class I EPSPS genes, and do not hybridize to probes from Class I EPSPS's. The Class II EPSPS enzymes are characterized by being more kinetically efficient than Class I EPSPS's in the presence of glyphosate. Plants transformed with Class II EPSPS genes are also disclosed as well as a method for selectively controlling weeds in a planted transgenic crop field.

Abstract: The present invention relates to a method of conveniently and efficiently producing reduced coenzyme Q10 having excellent qualities which is useful in foods, functional nutritive foods, specific health foods, nutritional supplements, nutrients, drinks, feeds, animal drugs, cosmetics, medicines, remedies, preventive drugs, etc. This method is suitable for industrial production thereof. In a method of synthesizing reduced coenzyme Q10 by reducing oxidized coenzyme Q10, followed by crystallization, at least one species selected from among hydrocarbons, fatty acid esters, ethers and nitrites is used as a solvent. Thus, the reduced coenzyme Q10 can be protected from oxidation, and as a result, the formation of the oxidized coenzyme Q10 as a by-product can be minimized, thereby giving reduced coenzyme Q10 having excellent qualities.

Abstract: A process for preparing lysozyme is provided which is characterized by mixing the egg white or its diluted solution with diatomaceous earth, kaolin, zeolite, or the mixtures thereof, and followed by eluting the adsorbed lysozyme with a salt solution. The diatomaceous earth, kaolin, and zeolite can specifically adsorb the lysozyme in egg white, and the adsorbed lysozyme can be easily eluted with a salt solution. According to the process of the present invention, the lysozyme in egg white can be easily and effectively isolated.

Abstract: A method of extracting a polypeptide of interest from a fermentation broth comprising: i) adjusting the pH close to pI of the polypeptide of interest; ii) adding a non-ionic surfactant with a hydrophile-lipophile balance (HLB) of 12 or lower; iii) cooling the mixture for solubilization and incubating at above cloud point for extraction; iv) phase separating at below cloud point to obtain liquid-liquid-solid fractions; and v) recovering the surfactant-rich top phase containing the polypeptide of interest.

Abstract: The present invention provides methods of purification of Hepatitis A Virus from the supernatant of an infected cell culture and production of a preparation of purified HAV antigen. The present invention is also directed to an HAV vaccine composition comprising a preparation consisting of purified mature HAV particles in an amount sufficient to induce a protective immune response in a mammal.

Abstract: An enzyme in a mixture containing the enzyme and a contaminant enzyme is purified by selectively aggregating and precipitating the contaminant enzyme with a surfactant. A deacetylase contaminant is separated from Cephalosporin C acylase using a cationic surfactant in an amount of 0.1 to 0.6%. Surfactants include benzylated or methylated cationic surfactants such as alkyl(palm)dimethylbenzyl ammonium chloride, alkyl(palm)trimethyl ammonium chloride and dodecyltrimethyl ammonium chloride. An immobilized enzyme is regenerated using a protease to remove the enzyme from a carrier. The carrier may be a synthetic adsorbent or an ion exchange resin having pores of 100 nm or less in diameter, and the immobilized enzyme is optionally crosslinked. Immobilized cephalosporin C acylase is regenerated using an alkaline or acidic protease.

Abstract: The present invention provides a crystallization process wherein a starting temperature is selected such that a desirable crystal morphology (e.g., square) is obtained. A temperature shift is then introduced, providing that the shift is not enough to induce further nucleation, where the crystals continue to grow in the desirable fashion, but with different kinetics, e.g., a higher rate of crystallization. As a result, the process gives a crystalline product with desirable morphology at a higher crystallization rate.

Abstract: The utility of soybeans having a composition of greater than 40% of the protein as beta-conglycinin and less than 10% of the protein as glycinin for making highly functional high beta-conglycinin compositions was discovered. The discovered ingredients are useful for mimicking the texturizing properties of casein while also maintaining or improving physiological benefits of soy protein ingredients (e.g., cholesterol and triglyceride lowering properties). The high stability of the high beta-conglycinin compositions against protein—protein aggregation reactions is valuable for creating good tasting beverages and beverage mixes. Cheese with good spreadability, gloss and smoothness was made using an enzyme-modified version of the new ingredient composition. Cheese with good firmness and meltability was also created using a different enzyme-treatment. High beta-conglycinin compositions were found to demonstrate excellent emulsifying and gelling properties in the pH region (5.5-6.

Abstract: Detergent formulations are prepared by directly agglomerating a fermentation broth extract, containing a detergent-type enzyme and a nonionic detergent-type surfactant, with a suitable detergent base mixture, without need for prior isolation of the enzyme.

Abstract: The invention relates to a method for producing optically pure compounds of 3(R)- and 3(S)-hydroxy-1-methyl-4-(2,4,6-trimethoxyphenyl)-1,2,3,6-tetrahydro-pyridine or its carboxylic acid esters by reacting the enantiomer mixtures stereoselectively, using an enzyme.

Abstract: The present invention provides a crystallization process wherein a starting temperature is selected such that a desirable crystal morphology (e.g., square) is obtained. A temperature shift is then introduced, providing that the shift is not enough to induce further nucleation, where the crystals continue to grow in the desirable fashion, but with different kinetics, e.g., a higher rate of crystallization. As a result, the process gives a crystalline product with desirable morphology at a higher crystallization rate. The starting temperature of the process can be between about 4° C. and 20° C. for no more than about 5 hours and the temperature shift of the process can be between about 22° C. and 60° C. for no more than about 20 hours.

Abstract: An enzyme in a mixture containing the enzyme and a contaminant enzyme is purified by selectively aggregating and precipitating the contaminant enzyme with a surfactant. A carrier containing an immobilized enzyme is regenerated by using a protease to remove the enzyme from the carrier. Cephalosporin C acylase is purified from a mixture of the acylase and a deacetylase contaminant by selectively aggregating and precipitating the deacetylase contaminant by adding to the solution a benzylated cationic or methylated cationic surfactant in an amount of 0.1 to 0.6% of the mixture. The surfactant may be alkyl(palm)dimethylbenzyl ammonium chloride. A carrier containing immobilized cephalosporin C acylase is regenerated by contacting the carrier with an alkaline or acidic protease. The acylase is immobilized by being bound to the carrier and optionally crosslinked. The carrier may have pores of 100 nm or less in diameter.

Abstract: A process is disclosed for preparing a detergent powder containing enzymes where the enzymes are extracted from a fermentation broth with a salt and surfactant mixture and directly agglomerated with detergent paste and dried to form the detergent powder. The process results in a two phase system where the enzyme is extracted into the surfactant rich phase and the second phase is salt rich. The process is especially useful for whole or clarified fermentation broths.

Abstract: A method for purification, and isolation in crystalline form, of a cellulase from a broth comprises: treating the broth with a crystallization-effective amount of a water-miscible organic solvent (e.g. a lower aliphatic alcohol or ketone); and isolating the cellulase in question in crystalline form.

Abstract: The present invention relates to a method for crystallization of a protein obtained from a protein-containing solution which involves (a) treating the protein-containing solution with a salt containing a sulphur atom having an oxidation state less than 6, and (b) recovering the protein in crystalline form.

Abstract: A process is disclosed for treating compositions of alcohol oxidase to inactivate catalase therein, which comprises aging the composition comprising alcohol oxidase and catalase at a temperature and for a time period sufficient to inactivate catalase while maintaining the alcohol oxidase activity.

Abstract: Aqueous two phase systems are provided which permit the extractive separation of biological materials. In a preferred embodiment, an aqueous solution of arabinogalactan defines at least one phase, and an aqueous solution of a second solute, such as a polyether, defines at least one other phase. In one embodiment, an aqueous two phase system is provided in which one phase is defined by an aqueous solution containing predominantly ultrarefined arabinogalactan, and the other phase is defined by an aqueous solution containing predominantly a polymer such as a poly(ethylene glycol). Biological materials which can be extracted and separated from mixtures using the aqueous two phase systems include cells, organelles, macromolecules and organic molecules. The aqueous two phase systems can be used in wide range of different applications including in bioconversions for the production and separation of enzyme reaction products.

Abstract: Supercritical and near critical fluids are used to fractionate biomass materials in two steps. In the first step, the biomass is exposed to elevated pressure supercritical or near critical fluid to bring about disruption of the biomass. In the second step, the disrupted biomass is subjected to a multiplicity of supercritical or near critical fluid extraction steps, with different solvation conditions used for each fraction. Thus, fractionation of the biomass is effected.

Abstract: A method for rapidly and inexpensively crystallizing enzymes from an impure mixture is disclosed. The yield of the enzyme is greater than 35% within twelve hours. The crystallizing agent is a salt added in a concentration which is less than the concentration necessary to crystallize the enzyme in amorphous form.

Abstract: Particular enzymes are precipitated from a mixture of proteins by the simultaneous addition of a soluble aluminate and acid. The pH of the mixture is at least one pH unit from the isoelectric point of the particular enzyme.

Abstract: The present invention relates to a method of separating an enzyme from an aqueous solution comprising this enzyme in mixture with other proteins, and recovery of the desired enzyme on crystalline form.

Abstract: A process and apparatus for reducing the volume and mass of solid waste (A) by initially subjecting the solid waste to a digestive enzymatic solution (20) agitated by fluid jets (28) under conditions which convert substrate (12) into a liquid waste which is discharged through a conventional sewage system (30). Non-biodegradable plastic shells (10) may be collected in a strainer basket (24) to be removed for further solid waste treatment such as shredding and the like.

Abstract: Peroxidase can be recovered from seed hulls in an improved method using a freeze-thaw technique. First, the seed hulls are comminuted, placed into water and homogenized. Next, the homogenate is frozen then thawed. The enzyme is then recovered from the aqueous solution by conventional means. Soybean or rice seed hulls can be used in this process.

Abstract: A method for preparing an aqueous solution enriched in EG III from an aqueous mixture containing cellulase proteins, xylanase and EG III is disclosed. The method involves adding an amount of a low molecular weight alcohol selected from the group consisting of ethanol, methanol, propanol and mixtures thereof to the aqueous mixture containing cellulase proteins, xylanase and EG III and an organic salt under conditions wherein substantially all of the cellulase proteins other than EG III and xylanase are precipitated out of the aqueous mixture. The method then involves removing the precipitate from the aqueous mixture so as to recover an aqueous supernate enriched in EG III. Next, the method involves adding an amount of an inorganic salt to the supernate produced in step b) so as to form a second precipitate and a second supernate and then finally collecting the second supernate from the second precipitate to obtain a supernate enriched in EG III.

Abstract: A method of separating a hydrophobic fermentation product selected from the group consisting of lipase, esterase, endoxylanase and an antibiotic from a mixture comprising said product and contaminants which method comprises adding to the mixture sequentially(1) 0.5 to 15% (w/v) of a nonionic surfactant,(2) 0.5 to 60 mg of a flocculating agent per gram of said mixture,(3) 1 to 20% (w/v) of an extra nonionic surfactant,(4) a suitable K, Na, NH.sub.4 or Mg salt selected from the group of chlorides, sulfates, acetates, carbonates or phosphates, whereby the concentration of the salt is chosen so as to have the surfactant layer on top and is between 2 and 30%;to obtain a three phase product mixture, separating the product mixture into liquid-liquid-solid fractions and recovering the hydrophobic fermentation product.

Abstract: Disclosed herein are buffers and methodologies for the capillary zone electrophoretic analysis of hemoglobin and the variants of hemoglobin. In a particularly preferred embodiment, the buffer comprises at least about 100 mM of barbituric acid, derivatives of barbituric acid, combinations of barbituric acid and derivatives of barbituric acid, and combinations of the derivatives of barbituric acid, and has a pH of at least about 8.0.

Abstract: A highly purified alkaline protease preparation is produced by separating a mixture containing amorphous form and crystalline form of alkaline protease from ultrafiltrate containing impurities, through a method which includes the incubation of an aqueous alkaline protease concentrate with sodium chloride and carbohydrate hydrolyse enzymes at an elevated temperature greater than 20.degree. C. and a pH between pH 4.0 and pH 6.5 at constant agitation.

Abstract: A method for preparing an aqueous solution enriched in xylanase only, EG III only and both EG III and xylanase from an aqueous mixture containing cellulase proteins, xylanase and EG III is disclosed. The methods involve adding an amount of a low molecular weight alcohol and an organic salt to an aqueous mixture containing cellulase proteins under conditions wherein substantially all of the cellulase proteins other than EG III and xylanase are precipitated out of the aqueous mixture. The methods can then involve 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 either the second precipitate from the second supernate to obtain a precipitate enriched in xylanase or the second supernate from the second precipitate to obtain a supernate enriched in EG III.

Abstract: The present invention pertains to a method of purification of DNA polymerase I, and the polymerase and Nick-translation activities thereof. In one embodiment, the method of purification is directed to circumstances where there are amplified amounts of the same relative to that which is found naturally occurring. In another embodiment, the purification method is directed to shortening the time period for the purification of the same whether in an amplified amount or not as compared to the time taught in the prior art.

Abstract: A novel compound for producing a purified enzyme precipitate and a method of preparing the purified enzyme precipitate, wherein the compound is an organic compound selected from the group consisting of carboxylic acids having at least 2 carboxyl groups, salts or esters of these carboxylic acids, amino acids, salts or esters of these amino acids.

Abstract: The subject invention provides a method for recovering a solution containing purified, enzymatically active Cu-Zn superoxide dismutase or a polypeptide analog thereof having substantially the same amino acid sequence as, and the bioloical activity of, naturally-occurring Cu-Zn superoxide dismutase from a composition which comprises cells containing Cu-Zn superoxide dismutase or a polypeptide analog thereof. The invention also provides a method of increasing the yield of recovered solutions having an increased concentration of b isoform of an enzymatically-active polypeptide analog of Cu-Zn superoxide dismutase from a composition which comprises cells containing a, b and c isoforms of the polypeptide analog.

Abstract: A purified luciferase from Luciola lateralis is disclosed. The enzyme is characterized by having properties including: an optimum pH range of 7.5 to 9.5, an optimum temperature range of 0.degree. C. to 50.degree. C., and that the enzyme does not act on ADP, CTP, UTP, and GTP. The enzyme is purified by using a process which includes: gel filtration chromatography, hydroxyapatite column chromatography, and a tris(hydroxy)aminomethane-hydro-chloric acid buffer.

Abstract: A process and apparatus for reducing the volume and mass of solid waste (A) by initially subjecting the solid waste to a digestive enzymatic solution (20) agitated by fluid jets (28) under conditions which convert substrate (12) into a liquid waste which is discharged through a conventional sewage system (30). Non-biodegradable plastic shells (10) may be collected in a strainer basket (24) to be removed for further solid waste treatment such as shredding and the like.

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: 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: The present invention pertains to a method of purification of DNA polymerase I, and the polymerase and Nick-translation activities thereof. In one embodiment, the method of purification is directed to circumstances where there are amplified amounts of the same relative to that which is found naturally occurring. In another embodiment, the purification method is directed to shortening the time period for the purification of the same whether in an amplified amount or not as compared to the time taught in the prior art.

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: Disclosed are methods for the recovery and purification of naturally produced chymosin. In particular, disclosed are methods for the recovery and purification of chymosin from aqueous solutions containing chymosin, pepsin and other contaminants.

Abstract: Disclosed are methods for the recovery of microbially produced chymosin. In particular, disclosed are methods for the recovery of chymosin from the fermentation beer arising from culturing microorganisms which have been engineered so as to produce chymosin. Also disclosed are methods for the selective recovery and subsequent purification of microbially produced chymosin.

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: 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: 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: 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 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 process for the purification of alcohol oxidase from whole cells of Pichia pastoris grown on methanol by the sequential steps of autolysis, crossflow filtration, ultrafiltration and recrystallization.

Abstract: This invention relates to bovine rennin and prorennin in the form of refractile bodies.

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: 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.