TAILIEUCHUNG - Báo cáo khoa học: Deciphering enzymes Genetic selection as a probe of structure and mechanism

The efficient engineering of enzymes with novel activities remains an ongoing challenge. Towards this end, genetic selection techniques provide a method for finding rare solutions to catalytic problems that requires only a limited foreknowledge of structure–function relationships. We have used genetic selections to extensively probe the structure and mechanism of chorismate mutases. The insights gained from these investigations will aid future enzyme design efforts. | Eur. J. Biochem. 271 1630-1637 2004 FEBS 2004 doi MINIREVIEW Deciphering enzymes Genetic selection as a probe of structure and mechanism Kenneth J. Woycechowsky and Donald Hilvert Laboratorium fur Organische Chemie Swiss Federal Institute of Technology ETH-Hõnggerberg Zurich Switzerland The efficient engineering of enzymes with novel activities remains an ongoing challenge. Towards this end genetic selection techniques provide a method for finding rare solutions to catalytic problems that requires only a limited foreknowledge of structure-function relationships. We have used genetic selections to extensively probe the structure and mechanism of chorismate mutases. The insights gained from these investigations will aid future enzyme design efforts. Keywords chorismate mutase functional selection protein engineering protein folding. Introduction The incredible catalytic power of enzymes is well-documented 1 2 but its source remains elusive. Enzymes catalyze a vast array of reactions with high specificity under mild conditions 3 . These properties make enzymes potentially useful for organic synthesis 4 5 . Still our current understanding of protein structure-function relationships remains insufficient for the de novo design of enzymes with tailored catalytic activities 6 . Evolution provides through multiple rounds of random mutagenesis and selection a means to circumvent this problem. This process has produced the vast array of proteins found in nature. In the laboratory directed evolution offers a promising strategy for the thorough study of protein structure-function relationships and for producing novel proteins with properties favorable for diverse applications including catalysis 7 8 . Principles of genetic selection Natural evolution selects for the survival and reproduction of organisms. By introducing DNA libraries encoding potential enzymes into microorganisms such as bacteria this process can be harnessed in the laboratory .

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