Abstract: There is disclosed a purified multimerase having an indirect and a direct proteolytic activity, which converts vWF having a singlet structure to vWF having a satellite structure and is active in the presence of the serine protease inhibitor DFP or the calpain protease inhibitor Z-Leu-Leu-Tyr-CHN.sub.2, as well as a method of preparing the multimerase according to the invention.

Abstract: A dry chemistry dye indicator composition provides improved shelf life, stable color indication end point and capability for a system at near normal pH. The novel dry chemistry dye indication system comprises 3-Methyl-6-(sodium sulfonate)-benzothiazolinone-(2)-hydrazone (MBTH-S). A preferred dye systems are based on the dye couple (MBTH-S) and 8-anilino-1-naphthalenesulfonate (ANS), and the dye couple MBTH-S and N-(3-sulfopropyl)analine. These dye indicator systems are used in conventional blood chemistry test strips and are particularly preferred for indication of glucose in blood.

Abstract: A self-addressable, self-assembling microelectronic device is designed and fabricated to actively carry out and control multi-step and multiplex molecular biological reactions in microscopic formats. These reactions include nucleic acid hybridizations, antibody/antigen reactions, diagnostics, and biopolymer synthesis. The device can be fabricated using both microlithographic and micro-machining techniques. The device can electronically control the transport and attachment of specific binding entities to specific micro-locations. The specific binding entities include molecular biological molecules such as nucleic acids and polypeptides. The device can subsequently control the transport and reaction of analytes or reactants at the addressed specific micro-locations. The device is able to concentrate analytes and reactants, remove non-specifically bound molecules, provide stringency control for DNA hybridization reactions, and improve the detection of analytes. The device can be electronically replicated.

Abstract: Antibacterial compounds for Ralstonia solanacearum having substantially (S)-3-(3-indolyl) butanoic acid of a specified structure or its salt as active component can selectively inhibit the growth of Ralstonia solanacearum and suppress bacterial wilt dependably even if used at relatively lower concentration.

Abstract: Compounds of the formula ##STR1## or the alkyl (1-6C), alkenyl (1-6C), or arylalkyl (7-12C) amides or salts including the cycloamido forms thereof;whereinn is 1 or 2;whereinwhen n is 1, X is a mono- or disubstituted or unsubstituted hydrocarbyl (1-20C) moiety optionally containing 1 or 2 nonadjacent heteroatoms (O, S or N), and wherein said substitution is selected from the group consisting of halo, OR, and SR, wherein R is H or lower alkyl (1-4C); when n is 2, one X is as above defined and the other X is lower alkyl (1-4C);Y is selected from the group consisting of ##STR2## wherein m is 1 or 2; andAA.sub.C is an amino acid coupled through a peptide bond to the remainder of the compound of formula 1, are useful as affinity ligands, elution reagents, solution inhibitors, diagnostic reagents and therapeutics. These compounds and analogous tripeptide glutathione analogs can be used as members of panels to obtain specific characteristic profiles for various glutathione-S-transferases.

Abstract: A process and device are provided for achieving cascade flow to achieve rapid separations of target components from fluid mixtures is provided. The devices of the invention have a cascading flow channel. The separation processes of the invention may include singly or in combination, the step of adjusting the total voids volume in all elements employed in the devices of the invention to be equal to or less than 6% of the total volume of fluid that is to be processed. A single separation process or a combination of separation processes may be used where the fluid mixture is whole blood, whole blood that is diluted or is in some other way treated, concentrated, divided into two or more streams, or augmented with blood or blood components, cellular materials such as proteins, antibodies, enzymes and fractions thereof, and nuclear materials such as DNA, RNA and their fragments.

Abstract: The invention relates to an immunogenic composition, characterized in that it comprises an adenyl cyclase-hemolysin (AC-Hly) protein, or an immunogenic portion of this AC-Hly, of a strain of Bordetella chosen from B. pertussis, B. parapertussis or B. bronchiseptica, and in that it comprises, in addition, a bacterial extract containing the expression products of the vrg genes of a strain of Bordetella chosen from B. pertussis, B. parapertussis or B. bronchiseptica, or a portion of these expression products which is sufficient to induce an immune response in a host to which the extract might be administered.

Abstract: An industrially applicable process for producing cis-3-hydroxy-L-proline, which is useful as a raw material for medicines or as an additive to foods. In the process, L-proline is converted into cis-3-hydroxy-L-proline in the presence of an enzyme source which is derived from a microorganism belonging to the genus Streptomyces or Bacillus and which catalyzes hydroxylation of L-proline into cis-3-hydroxy-L-proline, a divalent iron ion and 2-ketoglutaric acid, in an aqueous medium, and the produced cis-3-hydroxy-L-proline is collected from the aqueous medium. A novel enzyme L-proline-3-hydroxylase, a gene of L-proline-3-hydroxylase which is useful for the process, a transformant containing the gene, and a process for producing L-proline-3-hydroxylase using the transformant.

Abstract: Process for purifying serine proteases from a protein mixture by binding the serine protease to an immobilized polypeptide with the activity of an inhibitor DE-3 from Erythrina caffra, removing unbound components from the protein mixture, detaching the serine protease from the inhibitor and separating the immobilized inhibitor from the soluble serine protease and isolating serine protease which is characterized in that a polypeptide is used as the polypeptide which is the product of a prokaryotic or eukaryotic expression of an exogenous nucleic acid. This inhibitor is distinguished by an improved specific activity and is particularly suitable for the purification of plasminogen activators such as tissue plasminogen activators (t-PA and derivatives).

Abstract: A purified xylanase produced by Acidothermus cellulolyticus is disclosed having a pH optimum of between about 3.6-4.2 and a molecular weight of between about 50-55 kD as determined by gel filtration. The disclosed xylanase is useful in the bleaching of pulp for the production of paper and in treating feed compositions.

Abstract: Disclosed are improvements for enzyme-catalyzed reactions involving DNA or RNA, including restriction digests, which are based on conducting such reactions in the presence of lipids.

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: A novel method for immobilizing enzymes, while still keeping them in solution (thus under conditions of homogeneous catalysis), is reported. It consists in blocking enzymes in between two isoelectric membranes, having isoelectric points (pI) on either side of the pI of the enzyme to be "trapped". The reactor consists on a multichamber electrolyzer, in which the electric field is coupled to a hydraulic flow for continuously recycling the enzyme inside and outside the electric field to reservoirs acting as both heat exchangers and as feeders for injecting (or collecting) substrates, cofactors and other reagents. The pH of optimum activity is maintained by co-immobilizing the buffers within the enzyme reaction chamber. This is achieved by selecting appropriate amphoteric buffers, having a pI value comprised between the pI of the two membranes keeping the enzyme isoelectric and possessing a reasonable buffering power at their respective pIs.

Abstract: A method and system for separating a selected molecule from a heterogeneous mixture of molecules in aqueous solution are described. The method comprises (a) providing a separation device comprising a loading reservoir and a receiving reservoir coupled by a channel bearing immobilized microtubules aligned parallel to the longitudinal axis thereof the channel; (b) placing an aqueous solution containing the heterogeneous mixture of molecules in the loading reservoir; (c) adding a motor-ligand composition and ATP to the aqueous solution, wherein the motor-ligand composition comprises a motor protein for attaching to microtubules and moving therealong in the presence of ATP and the ligand is capable of binding the selected molecule, such that the ligand binds the selected molecule to form a complex and the complex moves along the immobilized microtubules to the receiving reservoir; and (d) removing the selected molecule from the receiving chamber.

Abstract: This invention provides a method for killing cells in fermentation mixtures in order to prepare the fermentation mixture for processing to recover or extract a desired product from the fermentation mixture. A preferred method of this invention comprises in either order, adjusting the pH of the fermentation mixture to a value equal to or less than about two pH units below the pK.sub.a of the compatible organic acid using a mineral acid, and adding a sufficient amount of a compatible organic acid and/or organic acid salt to the mixture to effect a substantially complete cell kill. The method of this invention is useful for killing microorganisms such as yeast, bacteria or fungi in any culture or fermentation mixture and is particularly useful in systems where it is desired to kill the cells without lysing them.

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: The subject invention pertains to a novel assay device for detecting oxalate in a sample. The assay device comprises enzyme and dye compositions immobilized on a solid carrier matrix. The subject invention can also be used to measure the concentration of oxalate in a sample. The subject invention further pertains to a novel oxalate oxidase composition and methods of preparing the subject enzyme composition. The oxalate oxidase composition can be used in the assay device of the subject invention.

Abstract: A dry chemistry dye indicator composition provides improved shelf life, stable color indication end point and capability for a system at near normal pH. The novel dry chemistry dye indication system comprises 3-Methyl-6-(sodium sulfonate)-benzothiazolinone-(2)-hydrazone (MBTH-S). A preferred dye systems are based on the dye couple (MBTH-S) and 8-anilino-1-naphthalenesulfonate (ANS), and the dye couple MBTH-S and N-(3-sulfopropyl)analine. These dye indicator systems are used in conventional blood chemistry test strips and are particularly preferred for indication of glucose in blood.

Abstract: Device for the treatment of nucleic acids from the sample, comprising a first reaction chamber for separating the nucleic acids from other sample components, and a second reaction chamber for the amplification of the nucleic acids, connected to said first reaction chamber via a controllable transport path.

Abstract: A method is provided for preparing an asparaginyl endoproteinase from the seeds of soybean, ginkgo and rice which have been collected between an early growing stage and ripening. The method comprises the steps of dialyzing an extract of the seeds against an acidic buffer of pH 4.0 to 6.0, ammonium sulfate precipitation, hydrophobic chromatography and gel filtration. The resulting asparaginyl endoproteinase cleaves glycinin between the C-terminal amino acid residue of the acidic subunit region, Asn, and the N-terminal amino acid residue of the basic subunit region, Gly or Asn.

Abstract: Disclosed is a nucleic acid segment encoding endothelin converting enzyme-2 (ECE-2), that produces mature ET-1 from big ET-1 both in vitro and in transfected cells. Also disclosed is a polypeptide composition comprising partially purified ECE-2 and methods of using ECE-2 to screen candidate substances as inhibitors of ECE-2. Methods of producing recombinant ECE-2 are also disclosed.

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 a protein, in particular an enzyme, from an aqueous solution of proteins, comprising (a) providing an aqueous mixture of proteins with a salt concentration at or below 1.5 Molar, to which a water soluble polymer has been added, and (b) recovery of the protein on crystalline form.

Abstract: An apparatus and method for selectively attracting and inhibiting attraction of at least one predetermined molecule to a site in a molecular detection device utilizes a first electrode and a second electrode proximate to the site. The first electrode selectively generates a first electric field proximate to the site in response to a first signal applied thereto. The first electric field provides an attractive force to attract the at least one predetermined molecule toward the site. The second electrode selectively generates a second electric field proximate to the site in response to a second signal applied thereto. The second electric field selectively inhibits attraction of the at least one predetermined molecule toward the site by providing a repulsive force which dominates the attractive force provided by the first electric field.

Abstract: Compositions containing two species of indolyl-3-alkane alpha-hydroxylase (INDH) are isolated from Pseudomonas XA. An INDH1 composition contains protein subunits having molecular weights of 75,000, 34,500 and 32,500 daltons. An INDH2 composition contains protein subunits having molecular weights of 60,000, 44,000 and 42,000 daltons. Molecular weights are determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The compositions have a Specific INDH Activity of at least 10 international Units of INDH activity per milligram of protein, and contain less than 1 nanogram of endotoxin per International Unit of Specific INDH Activity. The INDH compositions may be immobilized on an insoluble matrix such as silica beads to provide at least 2.5 international Units of INDH activity per gram of Immobilized INDH composition. The INDH compositions are isolated by lysing Pseudomonas XA cells at a temperature of no more than 15.degree. C.

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: This invention is within the field of thermostable proteases. More specifically, the present invention relates to a thermostable protease from Thermococcus celer, Thermococcus stetteri or Thermococcus litoralis, to a process for the preparation of these enzymes, and to detergent compositions comprising these enzymes. The enzyme has a temperature optimum in the range of from 75.degree. to 100.degree. C. and a pH optimum in the range of from 6.0-10.

Abstract: A self-addressable, self-assembling microelectronic device is designed and fabricated to actively carry out and control multi-step and multiplex molecular biological reactions in microscopic formats. These reactions include nucleic acid hybridization, antibody/antigen reaction, diagnostics, and biopolymer synthesis. The device can be fabricated using both microlithographic and micromachining techniques. The device can electronically control the transport and attachment of specific binding entities to specific micro-locations. The specific binding entities include molecular biological molecules such as nucleic acids and polypeptides. The device can subsequently control the transport and reaction of analytes or reactants at the addressed specific microlocations. The device is able to concentrate analytes and reactants, remove non-specifically bound molecules, provide stringency control for DNA hybridization reactions, and improve the detection of analytes. The device can be electronically replicated.

Abstract: A method for preparing an active and inactive adenylate cyclase from a culture of Bordetella parapertussis having specific reactivity with polyclonal or monoclonal antibodies to adenylate cyclase from Bordetella pertussis is presented. The inactive adenylate cyclase is devoid of calmodulin-activatable adenylate cyclase activity and of affinity for calmodulin. The method comprises culturing a clone of Bordetella parapertussis, homogenizing the culture to produce a homogenate, and isolating the active and inactive adenylate cyclase from the homogenate by urea precipitation. The isolated active and inactive adenylate cyclase may be used to prepare a vaccine for the prevention of Bordetella infection.

Abstract: A gene is provided encoding a DNA sequence for the expression of parathion hydrolase and a methionine analog thereof. In a preferred embodiment, a parathion hydrolase gene encoding sequence is obtained in which twenty-eight amino acids are deleted from the N-terminal amino acid sequence of parathion hydrolase. Parathion hydrolase or its analog is produced by transforming a host microorganism such as Escherichia, Bacillus or Streptomyces with the gene, culturing the microorganism in a culture medium and purifying parathion hydrolase therefrom. Highly purified soluble parathion hydrolase is produced by a purification method without using detergents such as Triton X-100 and Tween 20. Parathion hydrolase of enhanced activity is produced by adding cobalt or zinc or a mixture thereof to the culture medium in which the host microorganism is cultured.

Abstract: Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide amplification reaction. The devices are provided with a substrate microfabricated to include a polynucleotide amplification reaction chamber, having at least one cross-sectional dimension of about 0.1 to 1000 .mu.m. The device also includes at least one port in fluid communication with the reaction chamber, for introducing a sample to the chamber, for venting the chamber when necessary, and, optionally, for removing products or waste material from the device. The reaction chamber may be provided with reagents required for amplification of a preselected polynucleotide. The device also may include means for thermally regulating the contents of the reaction chamber, to amplify a preselected polynucleotide. Preferably, the reaction chamber is fabricated with a high surface to volume ratio, to facilitate thermal regulation.

Abstract: This invention relates to an apparatus containing specific binary combinations of glycosyltransferases, for the synthesis of specific saccharide compositions such as, for example, oligosaccharides, polysaccharides, glycolipids, and glycopeptides.

Abstract: Echinocandin B deacylase is purified to near homogeneity from Actinoplanes utahensis by a process comprising, in order, extracting the soluble enzyme, heating to 60.degree. C., hydrophobic interaction chromatography, (NH.sub.4).sub.2 SO.sub.4 fractionation, gel filtration, cation exchange chromatography, dye-ligand chromatography, gel filtration, and cation-exchange chromatography.

Abstract: A dipeptidylaminopeptidase (dDAP enzyme) and a method for isolating it from the culture medium of the cellular slime mold, Dictyostelium discoideum have been presented. The isolated dDAP enzyme has a pH optimum of about 3.5 and a mass of about 225 kDA. The dDAP enzyme has an activity which is somewhat similar to both DAP-I and DAP-III from this organism. Methods for using the dDAP enzyme to remove dipeptides from the N-terminus of recombinantly produced precursor proteins or peptides are also presented.

Abstract: The present invention relates to a method of determining histamine content as a freshness index of food. An examination liquid is injected in a reaction cell, an amount of dissolved oxygen (DO) is recorded through an oxygen sensor and an amplifier in the recorder. Then, an enzymatic reagent having histamine oxidase activity is injected in the reaction cell, a decrease in the dissolved oxygen is recorded in the recorder, and the histamine concentration is determined on the basis of the decrease by a micro computer.

Abstract: A method which uses a combination of polymeric cationic and an anionic flocculants to clarify an aqueous solution containing cells of a microorganism and parts thereof is described. The anionic flocculant is based upon a copolymer of an alpha-beta unsaturated monomer of an anhydride, a carboxylic acid or salt and an alpha-beta Unsaturated sulfonic acid or salt monomer. The polymeric cationic surfactant is preferably a quaternary ammonium or tertiary amine containing polymer produced from an alpha-beta unsaturated amino monomer. The method is particularly useful for clarifying solutions wherein a bacterium is used to produce an expressed material such as a protein, peptide, or amino acid dissolved in the solution which is to be separated from the cells or cell parts.

Abstract: Process for the production and extraction of thermostable super-oxide-dismutases from a photosynthetic microorganism cell. The thermostable superoxide-dismutase production and extraction process consists a) of culturing in the temperature range 40.degree. to 80.degree. C., in a closed photoreactor made from a light-transparent material and which is thermally resistant within said range, aerobic, photosynthetic, thermophilic microorganisms, which produce oxygen and grow exponentially in said range, said microorganisms being suspended in a culture medium and chosen from among microalgae and cyanobacteria and b) extracting from the culture medium the freshly produced, thermostable superoxide-dismutases, by cellular crushing, ultrafiltration and selective precipitation.

Abstract: An ultra-pure, clear thrombin solution having a high specific activity is described as well as a method of manufacture.

Abstract: A chromatographic process for the copurification of chondroitinase proteins useful in ocular surgery for non-surgical disruption of chondroitin sulfate, the molecule which mediates the attachment between the retina and vitreous body in the human eye. The process involves the use of ion exchange resins in conjunction with an affinity elution with chondroitin sulfate to afford copurification of the proteins.

Abstract: A Coffea canephora-D-galactosidase isozyme is purified by extracting a supernatant from Coffea beans containing the isozyme, extracting tannin from the supernatant, and isolating the isozyme. Preferrably, the supernatant is extracted by exposing the supernatant to insoluble polyvinylpolypyrrolidone in an amount sufficient to remove the tannin from the supernatant by forming hydrogen bonds with the tannin.

Abstract: A new restriction endonuclease capable of recognizing the nucleotide sequence of the following formula 1 in double stranded DNA. ##STR1## and specifically cleaving it is disclosed. The endonuclease may be made cultivating a strain of the genus Streptomyces (FERM BP-4836) capable of producing it.

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. A method for recovering a solution of purified human pancreatic cholesterol esterase having a molecular weight of about 100,000 daltons is disclosed. The 100,000 dalton fraction of human pancreatic cholesterol esterase is purified first, by preparing a solution of dialyzed human pancreatic cytosol containing the esterase and other proteins. Next, the solution is passed over a hydroxyapetite column and then eluted to yield all of the molecular weight fractions of the esterase and other proteins in the solution present as a single peak to give a first eluent.

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: This invention relates to an improved method of purifying the enzyme .alpha.-N-acetylgalactosaminidase from avian liver, as well as to the purified enzyme. The enzyme is capable of removing A antigens from the surface of cells in blood products. The method of purifying the enzyme is simple and time efficient, and lends itself to large scale production.

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 process is disclosed for the separation of an enantiomerically enriched 1-tosyloxy-2-acyloxy-3-butene and an enantiomerically enriched 1-tosyloxy-2-hydroxy-3-butene from a first mixture containing both compounds. The process includes the steps of:(a) forming a solution of the mixture in an organic solvent;(b) bringing the solution formed in (a) to a temperature wherein most of the enantiomerically enriched 1-tosyloxy-2-hydroxy-3-butene precipates, leaving in solution most of the enantiomerically enriched 1-tosyloxy-2-acyloxy-3-butene; and(c) separating the precipitate formed in (b) from the solution.

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: A crude membrane fraction of leaf sheath tissue of a grass endophytically infected by A. typhinum is produced. The extract exhibits endoproteolytic activity in the presence of detergent or after methanol precipitation. This activity is optimal at 35.degree.-40.degree. C. and pH 10-11 and is exhibited only when a reductant is present. The activity is associated with a first gel electrophoresis band having apparent molecular weight of 205,000 daltons when said extract is electrophoresed without prior boiling; however, when said extract is boiled prior to electrophoresis, there results a band having apparent molecular weight of 34,000 daltons. This band gives rise to polyclonal rabbit antibodies which are not cross react with proteinase K. The preferred source of Acremonium typhinum is ATCC 74228.

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.