Improving enzyme properties: when are closer mutations better?
Study of mutations that improve enzyme properties reveals that in many, but not all, cases closer mutations are more effective than distant ones. For enantioselectivity, substrate selectivity and new catalytic activity (catalytic promiscuity) closer mutations improved enzymes more effectively than distant ones. However, both close and distant mutations can improve activity, thermal stability and also probably stability toward organic solvents. Typical random mutagenesis methods, such as error-prone PCR, create greater numbers of distant mutations than close mutations because enzymes contain more amino acids distant from the active site than close to the active site. This suggests that instead of mutating the entire enzyme, focusing mutations near the substrate-binding site might dramatically increase the success rate in many directed evolution experiments.
Morley
Krista L
KL
Department of Chemistry, McGill University, 801 Sherbrooke Street West, Montréal, Québec H3A 2K6, Canada.
Kazlauskas
Romas J
RJ
D000818:Animals; D004798:Enzymes / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D016296:Mutagenesis; D013379:Substrate Specificity / Q000235:genetics
D004798:Enzymes