Abstract: A ubiquitin hydrolase is provided having a purity of at least 70% homogeneity based on the weight of the total protein in the composition. Also provided are DNA sequences encoding ubiquitin hydrolases, as well as expression systems for their recombinant production. Processes are provided for purification of a ubiquitin hydrolase from eukaryotes and for its use in recovering any desired polypeptide free from its fusion at its N-terminus with ubiquitin.
Abstract: A process for the one-step conversion of cephalosporin C and derivatives thereof to the corresponding 7-aminocephalosporanic acid and derivatives comprising treating said cephalosporin C and derivatives with a cephalosporin C amidase enzyme of a recited sequence, the DNA encoding said enzyme, and expression thereof in a suitable host, e.g., Bacillus species under the control of a suitable promoter.
Type:
Grant
Filed:
June 27, 1989
Date of Patent:
April 14, 1992
Assignee:
Merck & Co., Inc.
Inventors:
Mark S. Crawford, David B. Finkelstein, John A. Rambosek
Abstract: A gene which provides resistance to at least one methionine derivative and is capable of enhancing accumulation of S-adenosyl-L-methionine (SAM) in a cell, a hybrid plasmid having the same, a cell transformed with the above hybrid plasmid, and a process for producing SAM using the above cell. According to the present invention, SAM, which has various therapeutic effects, can be producing in a large amount at a low cost.
Abstract: Methods and compositions are provided for endopeptidase production, enhanced efficiencies of processing in vivo and in vitro to provide for process polypeptides, and purified enzyme for in vitro processing of polypeptides. The endopeptidase is specific for dibasic amino acid sites, cleaving at the C-side of the dipeptide.The S. cerevisiae strain pYBCA-5 (truncated KEX2) was deposited at the A.T.C.C. on June 21, 1984 and given Accession No. 20717.
Type:
Grant
Filed:
April 8, 1988
Date of Patent:
December 31, 1991
Assignees:
Chiron Corporation, The Regents of the University of California
Inventors:
Anthony J. Brake, Lindley C. Blair, David Julius, Jeremy W. Thorner
Abstract: There is disclosed a protease having the following enzymatic properties in view of:(a) function and substrate specificity,(b) optimum pH,(f) activation, and(g) inhibition;(a) the protease can hydrolyze, in particular, a peptide bond on the C-terminal side of Y of a peptide X--Y--, in which X is Arg, Lys or Pro optionally having a peptide bond on the N-terminal side, Y is Arg and -- indicates a peptide bond;(b) the protease has an optimum pH of about 7.0 in Tris hydrochloride buffer;(f) the protease is activated with calcium chloride or a surfactant; and(g) the protease is inhibited with p-amidinophenylmethanesulfonyl fluoride, p-chloromercuribenzoic acid, a metal chelater, tetraacetic acid or a heavy metal.
Abstract: A method is provided for expressing the proteases dipeptidyl aminopeptidases A and B which are useful for processing precursor proteins. Genes controlling these proteases may be insdrted into appropriate vectors and transformed into cultures. The proteases may either be utilized to process precursor proteins in vivo or may be extracted from cultures and used to process precursor proteins in vitro.
Type:
Grant
Filed:
February 29, 1984
Date of Patent:
December 11, 1990
Assignee:
The Board of Regents of the Univ. of California
Inventors:
George Sprague, Ira Herskowitz, Jeremy Thorner, David Julius, Lindley Blair, Anthony Brake
Abstract: Cheese is prepared using rennin obtained from insoluble refractile bodies of a recombinant microbial host cell. The rennin is obtained by rupturing the recombinant hose cell, isolating and solubilizing the insoluble refractile bodies, and recovering active rennin. Recombinant techniques involve preparing cDNA corresponding to the coding sequence for calf rennin, introducing into an expression vector and expressing in a host cell. As much as 200 mg rennin per liter of culture may be recovered. Prorennin or preprorennin may be produced and rennin derived therefrom.
Type:
Grant
Filed:
June 3, 1987
Date of Patent:
October 9, 1990
Assignee:
Genentech, Inc.
Inventors:
Kirk J. Hayenga, Virgil B. Lawlis, Bradley R. Snedecor
Abstract: Rennin for making cheese is obtained from insoluble refractile bodies of a recombinant microbial host cell. The rennin is obtained by rupturing the recombinant host cell, isolating and solubilizing the insoluble refractile bodies, and recovering active rennin. Recombinant techniques involve preparing cDNA corresponding to the coding sequence for calf rennin, introducing into an expression vector and expressing in a host cell. As much as 200 mg rennin per liter of culture may be recovered. Prorennin or preprorennin may be produced and rennin derived therefrom.
Abstract: A process for producing protease by cultivating a protease-producing mold in a liquid medium, which is characterized by continuously adding a liquid medium containing a protein material to the culture medium after the substantial termination of proliferation of the mold cells.
Type:
Grant
Filed:
November 21, 1986
Date of Patent:
November 7, 1989
Assignee:
The Japanese Research and Development Association for Bioreactor System
Abstract: Disclosed is a new protease having the following properties: (1) it is able to hydrolitically cleave a peptide bond between two adjacent basic amino acids in a peptide chain; (2) it has a molecular weight of about 43,000 as determined by electrophoresis; (3) it is inhibited by phenylmethylsulphonyl fluoride and diisopropyl fluorophosphate, but is not inhibited by monoiodoacetate, p-chloromercuribenzoic acid, ethylenediaminetetraacetic acid, 1,10-phenanthroline, tosyl-L-lysine chloromethyl ketone, and leupeptin. The protease can be produced by culturing Saccharomyces cerevisiae, and recovering purification by conventional methods, and is useful as a processing enzyme for conversion of a prohormone to an active hormone.
Abstract: The present invention provides a process for the selective separation of endoproteases from aqueous solutions, wherein an aqueous solution containing proteases is treated with a complex, present in the solid phase, of alpha.sub.2 -macroglobulin with a divalent metal selected from zinc, cobalt, nickel and copper and the solid phase then separated off.The present invention also provides an agent for carrying out this process, wherein it comprises a solid carrier material which is loaded with a complex of alpha.sub.2 -macroglobulin and of a divalent metal selected from zinc, cobalt, nickel and copper.