Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1997-07-23
1999-08-31
Patterson, Jr., Charles L.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435 681, 435 691, 435471, 435476, 435483, 43525421, 4353201, 536 232, C12N 960, C12N 1511, C12N 1557, C12P 2100
Patent
active
059453290
ABSTRACT:
The invention provides customized proteases (i.e., mutant enzymes), methods of making customized proteases, as well as methods of using customized proteases. The customized proteases of the invention are derived from the known proteases. Altered transacylation reactions include the capability to perform transacylation reactions not substantially catalyzed by the known protease or the capability to perform transacylation reactions with improved yields, or both. The methods of the invention provide for customized proteases through site specific or random mutagenesis of the active site amino acids of the known proteases. The invention also provides for methods of using the customized proteases to prepare a preselected transacylation products. The preselected transacylation products produced can be modified by substitution at the N- or C-terminal with nucleophiles such as L-amino acids, D-amino acids, amino acid amides, and radioactive amino acids.
REFERENCES:
patent: 4339534 (1982-07-01), Johansen et al.
patent: 4806473 (1989-02-01), Johansen et al.
patent: 4959312 (1990-09-01), Sirotkin
patent: 5049656 (1991-09-01), Lewis et al.
patent: 5185258 (1993-02-01), Caldwell et al.
Olesen, et al. "The Activity of Carboxypeptidase Y toward Substrates with Basic P.sub.1 Amino Acid Residues Is Drastically Increased by Mutational Replacement of Leucine 178", Biochemistry, 33:11121-11126 (1994).
Watson, J.D,. et al, in The Molecular Biology of the Gene, "The genetic systems provided by E. Coli and its viruses (excerpt)", pp. 180-182 (1987).
Schwarz A., et al., Methods in Enzymology, vol. 184, "Enzymatic C-terminal Biotinylation of Proteins", pp. 160-166 (1990).
Carlsberg Res. Commun., vol. 50, 1985, pp. 273-284, J. Winther et al., "Increased Hydrophobicity of the S1' Binding Site in Carboxypeptidase Y Obtained by Site-Directed Mutagenesis".
BioMed., Biochim. ACTA, vol. 50, No. 10/11, 1991, pp. 157-162, J. Bongers, "Comparison of Enzymatic Semisythesis of Peptide Amides: Human Growth Hormone Releasing Factor and Analogs".
Biochemistry, vol. 33, 1994, pp. 508-517, U. Mortensen et al., "Site-Directed Mutagenesis on (Serine) Carboxypeptidase Y".
Protein Engineering, vol. 6, No. 8, 1993, pp. 927-937, R. Siezen et al., "Engineering of the Substrate-Binding Region of the Subtilisin-Like Cell-Envelope Proteinase of Lactococcus Lactis".
Alexander, et al., "Engineering the Zinc Binding Site of Human Carbonic Anhydrase II: Structure of the His-94-Cys Apoenzyme in a New Crystalline Form", Biochemistry, 32,,pp. 1510-1518, (1993).
Behravan, et al. "Fine tuning of the catalytic properties of human carbonic anhydrase II", Eur. J. Biochem., 195, pp. 393-396, (1991).
Behravan, et al., "Structural and functional differen3ces between carbonic anhydrase isoenzymes I and II as studied by site-directed mutagenesis", Eur. J. Biochem., 198, pp. 589-592, (1991).
Bracey, et al., "Spinach Carbonic Anhydrase: Investigation of the Zinc-Binding Ligands by Site-Directed Mutagenesis, Elemental Analysis, and EXAFS", Biochemistry, 33, pp. 13126-13131, (1994).
Chen, et al., "Interaction and influence of Phenylalanine-198 and Threonine-199 on Catalysis by Human Carbonic Anhydrase III", Biochemistry, 32, pp. 7861-7865, (1993).
Fierke, et al., "Functional Consequences of Engineering the Hydrophobic Pocket of Carbonic Anhydrase II", Biochemistry, 30, pp. 11054-11063, (1991).
Jewell, et al., "Enhancement of the Catalytic Properties of Human Carbonic Anhydrase III by Site-Directed Mutagenesis", Biochemistry, 30, pp. 1484-1490, (1991).
Kiefer, et al., "Engineering a Cysteine Ligand into the Zinc Binding Site of Human Carbonic Anhydrase II", Biochemistry, 32, pp. 9896-9900, (1993).
Krebs et al., "Determinants of Catalytic Activity and Stability of Carbonic Anhydrase II as Revealed by Random Mutagenesis", J. Bio. Chem., 268, pp. 948-952 (1993).
Krebs, et al., "Structural and Functional Importance of a Conserved Hydrogen Bond Network in Human Carbonic Anhydrase II", J. Bio. Chem., 268, pp. 27458-27466, (1993).
Krebs, et al., "Kinetic and Spectroscopic Studies of Hydrophilic Amino Acid Substitutions in the Hydrophobic Pocket of Human Carbonic Anhydrase II", Biochemistry, 32, pp. 4496-4505, (1993).
LoGrasso, et al., "Influence of Amino acid Replacement at Position 198 on Catalytic Properties of Zinc-Bound Water in Human Carbonic Anhydrase III", Biochemistry, 32, pp. 5786-5791, (1993).
Martensson, et al., "Characterization of Folding Intermediates of Human Carbonic Anhydrase II: Probing Substructure by Chemical Labeling of SH Groups Introduced by Site-Directed Mutagenesis", Biochemistry, 32, pp. 224-231, (1993).
Ren, et al., "Same properties of site-specific mutants of human carbonic anhydrase II having active-site residues characterizing carbonic anhydrase III", J. Biochem., 201, pp. 417-420, (1991).
Taoka, et al., "Comparison of Intra-and Intermolecular Proton Transfer in Human Carbonic Anhydrase II", J. Bio. Chem., 269, pp. 17988-17992, (1994).
Taoka, et al., "Catalysis by mutants of human carbonic anhydrase II: effects of replacing hydrophobic resides 198 and 204", Biochimia et Biophysica Acta, 1159, pp. 274-278, (1992).
Tu, et al., "Interactions of Active-site Residues and Catalytic Activity of Human Carbonic Anhydrase III", J. Bio. Chem., 37, pp. 23002-23006, (1994).
Tu, et al., "Kinetic Analysis of a Mutant (His-Tyr) Responsible for Human Carbonic Anhydrase II Deficiency Syndrome", J. Bio. Chem., 268, pp. 4775-4779, (1993).
Tweedy, et al., "Structure and Energetics of a Non-Proline cis-Peptidyl Linkage in a Proline-202-Alanine Carbonic Anhydrase II Variant", Biochemistry, 32, pp. 10944-10949, (1993).
Endrizzi et al., Biochemistry 33:11106 (1994).
Blachyl-Dyson et al., "Yeast Carboxypeptidase Y Can Be Translocated and Glycosylated Without its Amino-terminal Signal Sequence", J. Cell Biol., 104:1183-1191 (1987).
Bongers et al., "Semisynthesis of Human Growth Hormone-Releasing Factor by Trypsin Catalyzed Coupling of Leucine Amide to a C-Terminal Acid Precursor" Int. J. Peptide Protein Res., 40:268-273 (1992).
Botstein et al., "Sterile Host Yeasts (SHY): A Eukaryotic System of Biological Containment for Recombinant DNA Experiments", Gene, 8:17-24 (1979).
Breddam et al., "Semisynthesis of Human Insulin Utilizing Chemically Modified Carboxypeptidase Y", Carlsberg Res. Comm., 49:463-472 (1984).
Breddam et al., "Chemically Modified Carboxypeptidase Y with Increased Amidase Activity", Carlsberg Res. Comm., 49:535-554 (1984).
Breddam et al., "Determination of C-Terminal Sequences by Digestion with Serine Carboxypeptidases: The Influence of Enzyme Specificity", Carlsberg Res. Commun., 52:55-63 (1987).
Breddam et al., "Amidation of Growth Hormone Releasing Factor (1-29) by Serine Carboxypeptidase Catalyzed Transpeptidation", Int. J. Peptide Res., 37:153-160 (1991).
Busby et al., "Isolation of Mutant Promoters in the Escherichia coli Galactose Operon Using Local Mutagenesis on Cloned DNA Fragments", J. Mol. Biol., 154:197-209 (1982).
F. Dal Degan et al., "Purification and Characterization of Two Serine Carboxypeptidase from Aspergillus niger and Their Use in C-terminal Sequencing of Proteins and Peptide Synthesis", Appl. Environ. Microbiol., 58:2144-2152 (1992).
Dierks et al., "Three Regions Upstream from the Cap Site Are Required for Efficient and Accurate Transcription of the Rabbit beta-Globin Gene in Mouse 3T6 Cells", Cell, 22:659-706 (1983).
Dodson et al., "Mutagenesis of Bacteriophage T7 in vitro by incorporation of O.sup.6 -methylguanine During DNA Synthesis", PNAS, 79:7440-7444 (1982).
Dower et al., "High Efficiency Transformation of E. coli by High Voltage Electroporation", Nucleic Acid Res., 16:6127 (1988).
Eadie et al., "Mechanism of Mutagenesis by O.sup.6 -methylguanine", Nature 308:201-203 (1984).
Flavell et al., "Site-Directed Mutagenesis: Generation of an Extracistronic Mutation in Bacteriophage Qbeta RNA", J. Mol. Biol., 89:255-272 (1974).
Grossberger et al., "Incorporation into DNA of the Base Analog 2-aminopurine by the Epstein-Barr Virus-Induced DNA Polymerase in vivo and in vitro", PNAS, 78:5689-587 (1981).
Hayashi, "Carboxy
Breddam Klaus
Kielland-Brandt Morten C.
Mortensen Uffe Hasbo
Olesen Kjeld Ove
Stennicke Henning Ralf
Carlsberg A/S
Moore William W.
Patterson Jr. Charles L.
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