Enzyme redesign

Ornstein, R. L. (1991)- Why timely bioremediation of synthetics may require rational enzyme redesign preliminary report on redesigning cytochrome P450cam for trichloroethylene dehalogenation. In On-Site Bioreclamation. Processes for Kenobiotic and Hydrocarbon Treatment, ed. R. E. Hinchee and R. F. Olfenbuttel, pp. 509-14. Boston Butterworth-Heinemann. 

Wagner, J. Lemer. R.A. Barbas. C.F.. III. Efficient aldolase catalytic antibodies that use the enamine mechanism of natural enzyme. Science 1995. 270. 1797-1800. Arnold. F.H. Combinatorial and computational challenges for biocatalyst design. Nature 2001, 409. 253-257. Penning, T.M. Jez, J.M. Enzyme redesign. Chem. Rev. 2001. 101. 3027-3046. 

Engineering Catalysis. The holy grail of enzyme redesign is the engineering of entirely new catalytic activities, a property which is often denoted as catalytic promiscuity [530,531]. The latter has been driven by the rapidly increasing number of crystal structures of proteins, which allow to understand the molecular details of their catalytic mechanism. In this context, it was possible to re-engineer the catalytic activities of well studied proteins to furnish switched activities or even completely novel functions, which are rarely found in Nature (Table 3.8). 

While wild-type PAMO was unable to convert 2-phenylcyclohexanone efficiently, all deletion mutants readily accepted this ketone as substrate. All mutants also displayed a similar thermostability when compared with the parent enzyme. The most active mutant (deletion of S441 and A442) was used for examining its enantioselective properties. It was found that the mutant preferably formed the (/ )-enantiomer of the corresponding lactone E = 100). While CHMO also shows a similar enantioselective behavior, this PAMO deletion mutant is a better candidate for future applications due to its superior stability. This clearly demonstrates that PAMO can be used as parent enzyme to design thermostable BVMO variants. It also illustrates that the available crystal structure of PAMO will be of great help for BVMO redesign efforts. ... 

Redesigned and Expanded Treatment of Enzyme Mechanisms NEW Mechanism Figures designed to lead students through these reactions step by step. The first reaction mechanism treated in the book, chymotrypsin, presents a refresher on how to follow and understand reaction mechanism diagrams. Twelve new mechanisms have been added, including lysozyme. 

Perham, R. N., Scrutton, N.S. Berry, A. (1991)- New enzymes for old redesigning the coenzyme and substrate specificities of glutathione reductase. Bioessays, 13, 515-25. 

Redesign of an Enzyme s Active Site KDPC Aldolase... 

G. MacBeath, P. Kast, and D. Hilvert, Redesigning enzyme topology by directed evolution, Science 1998, 279, 1958-1961. 

S. Oue, A. Okamoto, T. Yano, and H. Kaga-miyama, Redesigning the substrate specificity of an enzyme by cumulative effects of the mutations of non-active site residues,/. Biol. Chem. 1999, 274, 2344-2349. 

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