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@ARTICLE{JThiele:856114,
author = {J. Thiele, Martin and Davari, Mehdi D. and König, Melanie
and Hofmann, Isabell and Junker, Niklas and Mirzaei
Garakani, Tayebeh and Vojcic, Ljubica and Fitter, Jörg and
Schwaneberg, Ulrich},
title = {{E}nzyme-{P}olyelectrolyte {C}omplexes {B}oost the
{C}atalytic {P}erformance of {E}nzymes},
journal = {ACS catalysis},
volume = {8},
issn = {2155-5435},
address = {Washington, DC},
publisher = {ACS},
reportid = {FZJ-2018-05760},
pages = {10876–10887},
year = {2018},
abstract = {Understanding interactions between polymers and enzymes to
boost enzymatic activity is of high importance for
application of enzymes in multicomponent systems, such as
laundry, food, pharmaceuticals, or cosmetics. Proteases are
widely used in industries and increased performance in the
presence of polymers has been reported. Boosting of enzymes
activity by polymers and understanding of the molecular
principles is of high interest in biomedical and
biotechnological applications. A molecular understanding of
the boosting effect of poly(acrylic acid) (PAA) and
poly(l-γ-glutamic acid) (γ-PGA) for a nonspecific
subtilisin protease (Protein Database (PDB) ID: 1ST3) was
generated through biophysical characterization (fluorescence
correlation and circular dichroism spectroscopies,
isothermal titration calorimetry), molecular dynamics
simulations, and protease reengineering (site-saturation
mutagenesis). Our study revealed that enthalpically driven
interactions via key amino acid residues close to the
protease Ca2+ binding sites cause the boosting effect in
protease activity. On the molecular level electrostatic
interactions results in the formation of
protease-polyelectrolyte complexes. Site-saturation
mutagenesis on positions S76, I77, A188, V238, N242, and
K245 yielded an increased proteolytic performance against a
complex protein mixture (trademark CO-3; up to $∼300\%$
and $∼70\%)$ in the presence of PAA and γ-PGA. Being able
to fine-tune interactions between proteins and negatively
charged polymers through integrative use of computational
design, protein re-engineering and biophysical
characterization proved to be an efficient workflow to
improve protease performance.},
cin = {ICS-5},
ddc = {540},
cid = {I:(DE-Juel1)ICS-5-20110106},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551)},
pid = {G:(DE-HGF)POF3-551},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000449723900099},
doi = {10.1021/acscatal.8b02935},
url = {https://juser.fz-juelich.de/record/856114},
}