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Fine‐tuning the activity and stability of an evolved enzyme active‐site through noncanonical amino‐acids
Author(s) -
Wilkinson Henry C.,
Dalby Paul A.
Publication year - 2021
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.15560
Subject(s) - chemistry , substituent , saturated mutagenesis , stereochemistry , active site , residue (chemistry) , aromatic amino acids , mutagenesis , phenylalanine , side chain , amino acid , substrate (aquarium) , acceptor , hydrogen bond , thermal stability , helix (gastropod) , enzyme , organic chemistry , biochemistry , molecule , mutant , ecology , oceanography , physics , condensed matter physics , biology , snail , gene , geology , polymer
Site‐specific saturation mutagenesis within enzyme active sites can radically alter reaction specificity, though often with a trade‐off in stability. Extending saturation mutagenesis with a range of noncanonical amino acids (ncAA) potentially increases the ability to improve activity and stability simultaneously. Here, the authors report one of the first examples in which both catalytic activity and stability are simultaneously improved via site‐specific ncAA incorporation into a previously‐evolved transketolase (S385Y/D469T/R520Q) active site.