Improving a natural enzyme activity through incorporation of unnatural amino acids
Ugwumba, Isaac N; Ozawa, Kiyoshi; Xu, Zhi-Qiang; Ely, Fernanda; Foo, Jee; Herlt, Anthony; Coppin, Christopher Wayne; Brown, Sue; Taylor, Matthew C; Ollis, David; Mander, Lewis; Schenk, Gerhard; Dixon, Nicholas Edward; Oakeshott, John Graham; Jackson, Colin; Otting, Gottfried
The bacterial phosphotriesterases catalyze hydrolysis of the pesticide paraoxon with very fast turnover rates and are thought to be near to their evolutionary limit for this activity. To test whether the naturally evolved turnover rate could be improved through the incorporation of unnatural amino acids and to probe the role of peripheral active site residues in nonchemical steps of the catalytic cycle (substrate binding and product release), we replaced the naturally occurring tyrosine amino...[Show more]
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