Trypsin; Chymotrypsin Patents (Class 435/213)
-
Patent number: 5252725Abstract: The invention provides .alpha.-1-antichymotrypsin and protein preparations comprising human .alpha.-1-antichymotrypsin produced by E. coli cells transformed with a DNA sequence encoding human .alpha.-1-antichymotrypsin. The invention also provides methods for producing .alpha.-1-antichymotrypsin. The invention further provides analogues of .alpha.-1-antichymotrypsin that exhibit antichymotrypsin, anti-trypsin and anti-thrombin activity and methods of producing the analogues.Type: GrantFiled: July 24, 1991Date of Patent: October 12, 1993Assignee: The Trustees of the University of PennsylvaniaInventors: Harvey Rubin, Zhi M. Wang
-
Patent number: 5218137Abstract: Light activated acyl-enzymes of the formula: ##STR1## are disclosed. In the compounds of Formula (III), ENZ is an enzyme, X is O or S, Y is --NR.sub.3 R.sub.4, --OR.sub.5, or --SR.sub.5, and Z is a nucleophile. m is 0 to 3 and n is 1 or 2. Y is substituted on the ring at either or both of the 4 and 6 position.R.sub.1 and R.sub.2 are each independently H, C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl.R.sub.3 and R.sub.4 are each independently H, C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl, except that R.sub.3 and R.sub.4 are not simultaneously both H. R.sub.5 is C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl.Methods of using the acyl-enzymes and intermediates for making the acyl-enzymes are disclosed.Type: GrantFiled: March 25, 1992Date of Patent: June 8, 1993Assignee: Duke UniversityInventors: Ned A. Porter, John D. Bruhnke
-
Patent number: 5213977Abstract: A human serine protease expressed in cytotoxic killer cells, having a mass of about 25.8kD, and having the amino acid residues of the serine protease charge-relay catalytic mechanism conserved is provided. The protease can be produced by recombinant DNA technology. The cDNA is also provided.Type: GrantFiled: July 20, 1992Date of Patent: May 25, 1993Assignee: The Board of trustees of the Leland Stanford Junior UniversityInventors: Irving L. Weissman, Howard K. Gershenfeld
-
Patent number: 5114851Abstract: Light activated acyl-enzymes of the formula: ##STR1## are disclosed. In the compounds of Formula (III), ENZ is an enzyme, X is O or S, Y is --NR.sub.3 R.sub.4, --OR.sub.5, or --SR.sub.5, and Z is a nucleophile. m is 0 to 3 and n is 1 or 2. Y is substituted on the ring at either or both of the 4 and 6 position.R.sub.1 and R.sub.2 are each independently H, C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl.R.sub.3 and R.sub.4 are each independently H, C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl, except that R.sub.3 and R.sub.4 are not simultaneously both H. R.sub.5 is C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl.Methods of using the acyl-enzymes and intermediates for making the acyl-enzymes are disclosed.Type: GrantFiled: August 29, 1989Date of Patent: May 19, 1992Assignee: Duke UniversityInventors: Ned A. Porter, John D. Bruhnke
-
Patent number: 5093250Abstract: Claimed is a process for the production of optically active bicyclo[3.3.0]octanedione-carboxylic acid esters of Formula I ##STR1## wherein R.sub.1 and R.sub.2 jointly represent an oxygen atom, orR.sub.1 and R.sub.2 individually represent the residue OR.sub.4 where R.sub.4 means methyl- or ethyl-, andR.sub.3 is a straight or branched-chain alkyl group of 1-3 carbon atoms,comprising enantioselectively saponifying and decarboxylating with .alpha.-chymotrypsin an optically inactive, prochiral bicyclo[3.3.0]octanedionedicarboxylic acid diester of Formula II ##STR2## wherein R.sub.1, R.sub.2 and R.sub.3 have the meanings set forth above, and recovering a compound of Formula I.Type: GrantFiled: July 13, 1988Date of Patent: March 3, 1992Assignee: Schering AktiengesellschaftInventors: Karl Petzoldt, Werner Skuballa
-
Patent number: 5017489Abstract: Antibodies, nucleic acid sequences, and methods for inhibition of lysis for a novel serine esterase produced by both murine and human cytotoxic T lymphocytes. The serine esterase has an apparent molecular weight of approximately 28,000-31,000, as determined by SDS gel electrophoresis under reducing conditions, and trypsin-like activity. Inhibition of the esterase correlates with inhibition of the cells' cytolytic activity. Specific inhibition of the serine esterase is useful as a method for immunosuppression as well as for the inhibition of cytolytic activity of T lymphocytes, both in vivo and in vitro. The genes encoding the murine and human serine esterase are homologous.Type: GrantFiled: August 22, 1988Date of Patent: May 21, 1991Assignee: Massachusetts Institute of TechnologyInventors: Mark S. Pasternack, Herman S. Eisen
-
Patent number: 5002871Abstract: The present invention provides a membrane method for the enzymatic synthesis of peptides accomplished by shifting the chemical equilibrium that exists in a reaction mixture between charged or ionized reacting amino acids and uncharged or non-ionized peptide product in the presence of a proteolytic enzyme such as thermolysin. The equilibrium is shifted by diffusion of the unchanged peptide product across an ion-rejection membrane which removes the uncharged peptide from the reaction mixture and preferably the diffused uncharged peptide is quickly converted to a charged species that cannot back-diffuse into the reaction mixture so that the uncharged peptide is effectively "pulled" across the membrane. An enzymatic conversion of the uncharged species utilizing an esterase having proteolytic activity such as aminoacylase I is disclosed. Copermeating reactants can be separated from the product mixture and returned to the reaction mixture.Type: GrantFiled: July 28, 1987Date of Patent: March 26, 1991Assignee: The Coca-Cola CompanyInventor: Guillermo A. Iacobucci
-
Patent number: 4973555Abstract: Novel human serine proteases are provided characterized by expression in cytotoxic killer cells, being about 25.8kD, and having the amino acid residues of the serine protease charge-relay catalytic mechanism conserved. The proteases can be produced by recombinant DNA technology. The cDNA is also provided.Type: GrantFiled: December 31, 1986Date of Patent: November 27, 1990Assignee: The Board of Trustees of the Leland Stanford Junior UniversityInventors: Irving L. Weissman, Howard K. Gershenfeld
-
Patent number: 4973554Abstract: A water soluble ligand bound polymer has been synthesized and used for purification and stabilization of trypsin, an easily autodigestible enzyme. The affinity polymer was formed by copolymerizing N-acryloyl-m-amino-benzamidine, and acrylamide in the absence of oxygen. Bound trypsin could be easily eluted by either arginine or benzamidine. At low temperature (<5.degree. C.), the polymer solution was very stable and retained its high capacity for trypsin binding after 6 months of storage. Trypsin can also be stored in this system for extended periods. Combining the principles of affinity chromatography and ultrafiltration, a process has been developed, using this polymer, for purification of trypsin. The purification process also features provisions for the recirculation of the eluant as well as the macroligand.Type: GrantFiled: February 26, 1988Date of Patent: November 27, 1990Assignee: National Research Council of CanadaInventors: John H. Luong, Keith B. Male, An-Lac Nguyen
-
Patent number: 4963491Abstract: 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.Type: GrantFiled: January 27, 1989Date of Patent: October 16, 1990Inventors: Lars G. I. Hellgren, Viggo Mohr, Jan G. Vincent
-
Patent number: 4847202Abstract: Certain novel heterocyclic compounds, aromatic thioesters, their preparation, and their use in inhibiting serine proteases with chymotrypsin-like and elastase-like specificity and in the treatment of diseases such as emphysema which involve tissue proteolysis.Type: GrantFiled: March 26, 1986Date of Patent: July 11, 1989Assignee: Georgia Tech Research InstituteInventor: James C. Powers
-
Patent number: 4783410Abstract: A novel serine esterase produced by cytotoxic T lymphocytes is insolated and characterized. The protein appears to be membrane bound and has a reduced apparent molecular weight of about 28,000 daltons. Inhibition of the esterase correlates with inhibition of the cells' cytolytic activity. The serine esterase is useful in making antibody and as a target for the inhibition of cytolytic activity by T-lymphocytes, both in vivo and in vitro.Type: GrantFiled: June 28, 1985Date of Patent: November 8, 1988Assignee: Massachusetts Institute of TechnologyInventors: Mark S. Pasternack, Herman N. Eisen
-
Patent number: 4777250Abstract: Low molecular weight chymotrypsin analogs exhibiting the binding and catalytic groups of the real enzyme of the formula ##STR1## where D equals .alpha.-, .beta.- or .gamma.-cyclodextrin; P is X or (CH.sub.2).sub.n X, wherein n equals 0 to 2 and X equals S, NH, or 0; Q equals substituted phenyl with the carboxylate ion attached to the ortho position or equals (CH.sub.2).sub.n, wherein n equals 0 to 3; and R equals hydrogen --CH.sub.3 or --C.sub.2 H.sub.5.The enzyme models are useful as an analytical model mimicking the real enzyme, a stain remover, a laundry detergent additive, a food additive, and as a therapeutic agent for treating enzyme deficiencies.Type: GrantFiled: June 19, 1986Date of Patent: October 11, 1988Assignee: Northwestern UniversityInventors: Myron L. Bender, Valerian T. D'Souza
-
Patent number: 4738925Abstract: 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.Type: GrantFiled: April 29, 1985Date of Patent: April 19, 1988Assignee: Union Carbide CorporationInventors: Kavssery P. Ananthapadmanabhan, Errol D. Goddard
-
Patent number: 4725545Abstract: Certain novel aryl sulfonyl fluorides, their preparation, and their use in inhibiting serine proteases with chymotrypsin-like and elastase-like specificity.Type: GrantFiled: February 2, 1987Date of Patent: February 16, 1988Assignee: Georgia Tech Research CorporationInventor: James C. Powers
-
Patent number: 4716113Abstract: An improved process for preparing (4S)-6-fluoro-spiro-[chroman-4,4'-imidazolidine]-2',5'-dione (sorbinil) or its (2R)-methyl derivative (2-methylsorbinil) is disclosed herein, starting from p-fluorophenol in each instance. The final products obtained have known pharmaceutical value as agents for the control of certain chronic diabetic complications. Key steps concerned with the process involve converting p-fluorophenol into the appropriate .beta.-(4-fluorophenoxy)alkane halide, followed by amidoalkylation with N-benzoyl or N-(lower alkanoyl)-.alpha.-hydroxyglycine to form an intermediate 2-amidoalkylated derivative thereof, and then dehydration and spiroalkylation of said intermediate by treatment with a dehydrating agent and a base to yield a spiroalkylated azlactone compound.Type: GrantFiled: March 31, 1986Date of Patent: December 29, 1987Assignee: Pfizer Inc.Inventor: Frank J. Urban
-
Patent number: 4532212Abstract: Process for the dissolution of peptides and proteins in non-aqueous and mixed non-aqueous/aqueous solvents using crown ethers, solutions thereby obtained and their use. Purification of proteins and/or enzymes, modification and optimisation of the activity of enzymes. Use of peptide-crown ether complexes as membrane materials, use of enzyme-crown ether complexes as detergents.Type: GrantFiled: June 26, 1984Date of Patent: July 30, 1985Assignee: Shell Oil CompanyInventor: Barbara Odell
-
Patent number: 4507233Abstract: Colored proteins having a known molecular weight of 1800 to 321200 are prepared by coupling monomers of a colored protein having a known molecular weight or by coupling a colored protein having a known molecular weight with a colorless protein having a known molecular weight. At least two proteins are selected from the so prepared colored proteins and used as a colored molecular weight marker. This colored molecular weight marker is used for determination of the molecular weight of a protein having an unknown molecular weight, and can also be used as a reference protein for purification of a protein having a known molecular weight.Type: GrantFiled: April 18, 1983Date of Patent: March 26, 1985Assignee: Oriental Yeast Co., Ltd.Inventors: Hiraku Saito, Yasuo Suzuki, Katsumi Fujii, Kaoru Miyazaki, Takekazu Horio
-
Patent number: 4499186Abstract: A method for diagnosis of rheumatoid arthritis and related autoimmune diseases comprises blocking calcium ions contained in a blood sample, effecting hydrolysis of a selected substrate in the presence of .alpha..sub.2 -macroglobulin (.alpha..sub.2 M) from the blood sample, and determining the extent of hydrolysis of the substrate. Preferably, .alpha..sub.2 M in plasma is incubated with a hydrolyzable chromogenic substrate and the liberated chromogen is determined spectrophotometrically. A diagnostic kit is also provided.Type: GrantFiled: December 30, 1982Date of Patent: February 12, 1985Assignee: University of Illinois, Board of TrusteesInventors: Marius C. Teodorescu, Alexander M. Gaspar, Gregory T. Spear, Doina Ganea, John L. Skosey
-
Patent number: 4489159Abstract: A process for converting human des-B30-insulin, namely ##STR1## into h-In-Thr.sup.B30 esters through amidation with an L-threonine ester in a mixture of water and a water miscible solvent in the presence of trypsin and optionally an acid.Yields in excess of at least 90% are obtained by limiting water content to 10%-30% v/v of the reaction mixture.Type: GrantFiled: August 9, 1982Date of Patent: December 18, 1984Assignee: Novo Industri A/SInventor: Jan Markussen
-
Patent number: 4411989Abstract: The disclosed methods and devices for detecting or monitoring or identifying substances (such as chemical warfare agents) and residual environmental pollutants (such as pesticides) utilize the discovery that spectra (e.g. infrared absorption spectra) of an uninhibited enzyme (e.g., a cholinesterase) can differ from spectra of the same enzyme which has been complexed with the agent pollutant. For example, the infrared spectrum of uninhibited butyrylcholinesterase (BuChE) lacks a distinct absorption peak found at about 1000 cm.sup.-1 in the BuChE-Malathion spectrum. The enzyme is used to collect and concentrate the agent or pollutant, and the resulting complexed enzyme can then be analyzed (e.g., by infrared spectroscopy) and its spectrum compared to an uninhibited enzyme spectrum.Type: GrantFiled: August 13, 1981Date of Patent: October 25, 1983Assignee: Midwest Research InstituteInventor: Ann E. Grow
-
Patent number: 4303592Abstract: A new compound amidinophenylmethylsulfonylfluoride is formed and preferably used in the para isomer form as an irreversible inactivator of selected serine proteases.Type: GrantFiled: January 9, 1980Date of Patent: December 1, 1981Assignee: Center for Blood Research, Inc.Inventors: Richard Laura, David H. Bing
-
Patent number: 4264738Abstract: A process for purification of proteolytic enzymes which comprises a biospecific sorption on a sorbent comprising a product of interaction of a solid carrier which is an aminoderivative of a siliceous material, a ligand and a condensation agent; elution of the sorbed enzymes by salt buffers and/or organic solvents.Type: GrantFiled: August 1, 1979Date of Patent: April 28, 1981Inventors: Valentin M. Stepanov, Galina N. Rudenskaya, Valery K. Akparov, Anatoly V. Gaida
-
Patent number: 4252902Abstract: A process for purification of crude kallikrein utilizing a specific affinity of kallikrein to p-aminobenzamidine, which comprises bringing a solution, containing kallikrein and contaminated by other undesirable enzymes and proteins, into contact with a water-insoluble carrier combined with p-aminobenzamidine, exclusively eluting out the other enzymes and proteins from the carrier by the use of a buffer solution of lower NaCl concentration and then exclusively eluting out kallikrein by the use of a buffer solution of higher NaCl concentration.Type: GrantFiled: July 17, 1979Date of Patent: February 24, 1981Assignee: Toho Pharmaceutical Industries Co., Ltd.Inventors: Setsuro Fujii, Kiyoaki Noda, Akio Yokoshima
-
Patent number: 4251631Abstract: A cross-linked enzyme membrane is prepared by directly adsorbing enzymes into the pores of a microporous non-fibrous filter membrane made of a silica modified vinylchloride polymer and then cross-linking the enzyme with a bi-functional coupling agent whereby enzyme molecules are cross-linked to each other without chemically bonding the enzyme molecules to the membrane. The cross-linked enzyme membrane is used to carry out enzymatic reactions by passing a solution of substrate for the enzyme through the membrane under a pressure differential.Type: GrantFiled: February 1, 1979Date of Patent: February 17, 1981Assignee: Research Products Rehovot Ltd.Inventor: Shulamith Simon
-
Patent number: 4212946Abstract: 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.Type: GrantFiled: May 18, 1978Date of Patent: July 15, 1980Assignees: Toyo Soda Manufacturing Co., Ltd., (Zaidanhojin) Sagami Chemical Research CenterInventors: Yuji Nonaka, Kiyotaka Oyama, Heijiro Satoh
-
Patent number: 4212945Abstract: 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.Type: GrantFiled: May 18, 1978Date of Patent: July 15, 1980Assignees: (Zaidanhojin) Sagami Chemical Research Center, Toyo Soda Manufacturing Co., Ltd.Inventors: Yuji Nonaka, Kiyotaka Oyama, Heijiro Satoh