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@ARTICLE{Petrovic:836815,
author = {Petrovic, Dusan and Frank, David and Kamerlin, Shina C. L.
and Hoffmann, Kurt and Strodel, Birgit},
title = {{S}huffling {A}ctive {S}ite {S}ub-{S}tate {P}opulations
{I}mpacts {C}atalytic {A}ctivity: {T}he {C}ase of {G}lucose
{O}xidase},
journal = {ACS catalysis},
volume = {7},
issn = {2155-5435},
address = {Washington, DC},
publisher = {ACS},
reportid = {FZJ-2017-05857},
pages = {6188–6197},
year = {2017},
abstract = {Glucose oxidase has wide applications in the
pharmaceutical, chemical, and food industries. Many recent
studies have enhanced key properties of this enzyme using
directed evolution, yet without being able to reveal why
these mutations are actually beneficial. This work presents
a synergistic combination of experimental and computational
methods, indicating how mutations, even when distant from
the active site, positively affect glucose oxidase
catalysis. We have determined the crystal structures of
glucose oxidase mutants containing molecular oxygen in the
active site. The catalytically important His516 residue has
been previously shown to be flexible in the wild-type
enzyme. The molecular dynamics simulations, performed in
this work, allow us to quantify this floppiness, revealing
that His516 exists in two states: a catalytic and a
non-catalytic one. The relative populations of these two
sub-states are almost identical in the wild-type enzyme,
with His516 readily shuffling between them. In the glucose
oxidase mutants, on the other hand, the mutations enrich the
catalytic His516 conformation and reduce the flexibility of
this residue, leading to an enhancement of their catalytic
efficiency. This study stresses the benefit of active site
preorganization in respect to enzyme conversion rates by
reducing molecular reorientations needs. We further suggest
that the computational approach based on Hamiltonian replica
exchange molecular dynamics, used in this study, may be a
general approach to screening in silico for improved enzyme
variants involving flexible catalytic residues.},
cin = {ICS-6 / JARA-HPC},
ddc = {540},
cid = {I:(DE-Juel1)ICS-6-20110106 / $I:(DE-82)080012_20140620$},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551) /
Computational Enzyme Design $(jics69_20151101)$},
pid = {G:(DE-HGF)POF3-551 / $G:(DE-Juel1)jics69_20151101$},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000410005700072},
pubmed = {pmid:29291138},
doi = {10.1021/acscatal.7b01575},
url = {https://juser.fz-juelich.de/record/836815},
}