Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing oxygen-containing organic compound
Patent
1996-11-12
1998-06-23
Patterson, Jr., Charles L.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing oxygen-containing organic compound
4351721, 4351723, 435190, C12P 740, C12N 1500, C12N 904
Patent
active
057704104
ABSTRACT:
A method for modifying the specificity or efficiency of an enzyme, while retaining its catalytic activity, is disclosed. The method is characterized by selecting an enzyme, the tertiary structure of which is substantially known or deduced; identifying a single specificity or efficiency-related region of the enzyme; identifying or constructing unique restriction sites bounding the identified region in the DNA coding therefor; generating a DNA sequence which corresponds to at least a portion of the identified region, except that the nucleotides of at least one codon are randomized, using the generated DNA sequence to replace the original such sequence; expressing the DNA including the generated DNA sequence; and selecting for a desired modification so that the DNA coding therefor may be isolated; the randomized DNA being generated by means of a PCR assembly method. Enzyme generated using this method, and having enhanced specificity or efficiency, are also disclosed.
REFERENCES:
patent: 5098841 (1992-03-01), Ghisalba
Siderovski, D. P., et al. (1992) Nucl. Acids Res. 20(20), 5311-5320.
Spee, J. H., et. al. (1993) Nucl. Acid Res. 21(3), 777-778.
Dunn, C. et al. (1991) "Design and synthesis of new enzymes based on the lactate dehydrogenase framework" Phil. Trans. R. Soc. Lond. B, 332:177-184.
Kang, A. et al. (1991) "Linkage of recognition and replication functions by assembling combinatorial antibody Fab libraries along phage surfaces" Proc. Natl. Acad. Sci USA, 88:4643-4366.
Wilks, H. et la. (1990) "Designs for a Broad Substrate Specificity Keto Acid Dehdrogenase" Biochemistry, 29:8587-8591.
Holm, L. et al. (1990) "Random mutagenesis used to probe the structure and function of Bacillus stearothermophilus alpha-amylase" Protein Engineering, 3(3):181-191.
Knoth, K. et al. (1988) "Highly degenerate, inosine-containing primers specifically amplify rare cDNA using the polmerase chain reaction" Nucleic Acids Research, 16(22):10932.
Simon, E. et al. (1989) "D-Lactate Dehydrogenase --Substrate Specificity and Use as a Catalyst in the Synthesis of Homochiral 2-Hydroxy Acids" Applied Biochemistry and Biotechnology, 22:169-179.
Oliphant, A. et al. (1989) "An efficient method for generating proteins with altered enzymatic properties Application to B-lactamase" Proc. Natl. Acad. Sci. USA, 86:9094-9098.
Erlich, H. et al. (1989) "Principles and Applications for DNA Amplification" PCR Technology, Stockton Press New York pp. 61-70 (1989).
Innis, M et al. (1988) "DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA" Proc. Natl. Acad. Sci. Usa, 85:9436-9440.
Clarke, A. et al. (1986) "Site-directed mutagenesis reveals role of mobile arginine residue in lactate dehydrogenase catalysis" Nature, 324(18/25):699-702.
Barstow, D. et al. (1986) "Cloning, expression and complete nucleotide sequence of the Bacillus stearothermophilus L-lacte dehydrogenase gent" Gene, 46:47-55.
Winter, G. et al. (1982) "Redesigning enzyme structure by site-directed mutagenesis: tyrosyl tRNA synthetase and ATP binding" Nature, 299;756-758.
Langenbeck, U. et al. (1981) "Renal Transport of Aromatic Acids in Patients with Phenylketonuria" J. Inher, Metab. Dis., 4:69-70.
Katzen, H. et al. (1965) "Multiple Forms of Hexokinase in the Rat: Tissue Distribution, Age Dependency, and Properties" Proc. N.A.S., 54:1218-1225.
Wilks, H. M., et al. (1991) Current Opinion in Biotechnology 2, 561-567.
Elhawrani Ayman
Hart Keith William
Holbrook Joseph John
Wilks Helen Margaret
Genzyme Corporation
Gosz William G.
Patterson Jr. Charles L.
Stole Einar
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