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@PHDTHESIS{kraus:638201,
author = {kraus, yasemine},
title = {{B}iophysikalische {U}ntersuchung von an
{B}iotransformationbeteiligten {F}lavinmonooxygenasen},
school = {Christian - Albrechts - Universität zu Kiel},
type = {Dissertation},
reportid = {PUBDB-2025-04019},
pages = {1-345},
year = {2025},
note = {Dissertation, Christian - Albrechts - Universität zu Kiel,
2025},
abstract = {This work deals with the expression, purification,
characterization, and crystallization of ΔhFMO5,a variant
of the human flavin-dependent monooxygenase 5 (hFMO5)
without transmembraneregion. This modification enables the
expression and purification of soluble protein
withoutaddition of detergent, resulting in the isolation of
active protein saturated with FAD. Theaggregation prone
ΔhFMO5 was successfully expressed in a fermenter using the
strain E. coliRosetta 2 (DE3) pLysS, lysed by enzymatic cell
disruption and purified through affinity and sizeexclusion
chromatography. Characterization of ΔhFMO5 via mass
photometry reveals that insolution there is present a
mixture of different oligomeric states, with the tetrameric
form beingpredominant. The catalytic cycle of ΔhFMO5
includes an uncoupled metabolic pathway, in whichNADPH is
consumed to regenerate the FAD cofactor without substrate
turnover, leading to theformation of hydrogen peroxide.
Iterative modeling of the reaction kinetics of
ΔhFMO5demonstrates that substrate metabolism as a
second-order reaction proceeds more slowlycompared to the
uncoupled reaction. Activity measurements show that the
conversion ofphenylacetone to benzyl acetate is catalyzed by
ΔhFMO5 with a turnover rate of 5.1 $\%.$ N-acetylserotonin
was identified as a potential endogenous substrate playing a
role in the productionof melatonin and serotonin.},
cin = {EMBL-User},
cid = {I:(DE-H253)EMBL-User-20120814},
pnm = {6G3 - PETRA III (DESY) (POF4-6G3)},
pid = {G:(DE-HGF)POF4-6G3},
experiment = {EXP:(DE-H253)P-P12-20150101},
typ = {PUB:(DE-HGF)11},
url = {https://bib-pubdb1.desy.de/record/638201},
}