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@ARTICLE{Zhao:857897,
author = {Zhao, Jing and Frauenkron-Machedjou, Victorine Josiane and
Fulton, Alexander and Zhu, Leilei and Davari, Mehdi D. and
Jaeger, Karl-Erich and Schwaneberg, Ulrich and Bocola,
Marco},
title = {{U}nraveling the effects of amino acid substitutions
enhancing lipase resistance to an ionic liquid: a molecular
dynamics study},
journal = {Physical chemistry, chemical physics},
volume = {20},
number = {14},
issn = {1463-9084},
address = {Cambridge},
publisher = {RSC Publ.66479},
reportid = {FZJ-2018-06851},
pages = {9600 - 9609},
year = {2018},
abstract = {Understanding of the structural and dynamic properties of
enzymes in non-aqueous media (e.g., ionic liquids, ILs) is
highly attractive for protein engineers and synthetic
biochemists. Despite a growing number of molecular dynamics
(MD) simulation studies on the influence of different ILs on
wild-type enzymes, the effects of various amino acid
substitutions on the stability and activity of enzymes in
ILs remain to be unraveled at the molecular level. Herein,
we selected fifty previously reported Bacillus subtilis
lipase A (BSLA) variants with increased resistance towards
an IL (15 $vol\%$ 1-butyl-3-methylimidazolium
trifluoromethanesulfonate; [Bmim][TfO]), and also ten
non-resistant BSLA variants for a MD simulation study to
identify the underlying molecular principles. Some important
properties differentiating resistant and non-resistant BSLA
variants from wild-type were elucidated. Results show that,
in 15 $vol\%$ [Bmim][TfO] aqueous solution, $40\%$ and
$60\%$ of non-resistant variants have lower and equal
probabilities to form a catalytically important hydrogen
bond between S77 and H156 compared to wild-type, whereas
$36\%$ and $56\%$ of resistant variants show increased and
equal probabilities, respectively. Introducing positively
charged amino acids close to the substrate-binding cleft for
instance I12R is beneficial for the BSLA resistance towards
15 $vol\%$ [Bmim][TfO], likely due to the reduced
probability of [Bmim]+ cations clustering near the cleft. In
contrast, substitution with a large hydrophobic residue like
I12F can block the cleft through hydrophobic interaction
with a neighboring nonpolar loop 134–137 or/and an
attractive π–π interaction with [Bmim]+ cations. In
addition, the resistant variants having polar substitutions
on the surface show higher ability to stabilize the surface
water molecule network in comparison to non-resistant
variants. This study can guide experimentalists to
rationally design promising IL–resistant enzymes, and
contribute to a deeper understanding of protein–IL
interactions at the molecular level.},
cin = {IMET / JARA-HPC},
ddc = {540},
cid = {I:(DE-Juel1)IMET-20090612 / $I:(DE-82)080012_20140620$},
pnm = {581 - Biotechnology (POF3-581) / Towards Discovery of
Molecular Determinants Underlying Organic Solvent Resistance
of Enzymes: Large- $(jara0169_20170501)$},
pid = {G:(DE-HGF)POF3-581 / $G:(DE-Juel1)jara0169_20170501$},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:29578220},
UT = {WOS:000429205700059},
doi = {10.1039/C7CP08470F},
url = {https://juser.fz-juelich.de/record/857897},
}