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@ARTICLE{Kokorin:904482,
author = {Kokorin, Arsenij and Parshin, Pavel D. and Bakkes, Patrick
J. and Pometun, Anastasia A. and Tishkov, Vladimir I. and
Urlacher, Vlada B.},
title = {{G}enetic fusion of {P}450 {BM}3 and formate dehydrogenase
towards self-sufficient biocatalysts with enhanced activity},
journal = {Scientific reports},
volume = {11},
number = {1},
issn = {2045-2322},
address = {[London]},
publisher = {Macmillan Publishers Limited, part of Springer Nature},
reportid = {FZJ-2021-06052},
pages = {21706},
year = {2021},
abstract = {Fusion of multiple enzymes to multifunctional constructs
has been recognized as a viable strategy to improve
enzymatic properties at various levels such as stability,
activity and handling. In this study, the genes coding for
cytochrome P450 BM3 from B. megaterium and formate
dehydrogenase from Pseudomonas sp. were fused to enable both
substrate oxidation catalyzed by P450 BM3 and continuous
cofactor regeneration by formate dehydrogenase within one
construct. The order of the genes in the fusion as well as
the linkers that bridge the enzymes were varied. The
resulting constructs were compared to individual enzymes
regarding substrate conversion, stability and kinetic
parameters to examine whether fusion led to any substantial
improvements of enzymatic properties. Most noticeably, an
activity increase of up to threefold was observed for the
fusion constructs with various substrates which were partly
attributed to the increased diflavin reductase activity of
the P450 BM3. We suggest that P450 BM3 undergoes
conformational changes upon fusion which resulted in altered
properties, however, no NADPH channeling was detected for
the fusion constructs.},
cin = {IBG-1},
ddc = {600},
cid = {I:(DE-Juel1)IBG-1-20101118},
pnm = {2172 - Utilization of renewable carbon and energy sources
and engineering of ecosystem functions (POF4-217)},
pid = {G:(DE-HGF)POF4-2172},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:34737365},
UT = {WOS:000714702000001},
doi = {10.1038/s41598-021-00957-5},
url = {https://juser.fz-juelich.de/record/904482},
}