<div class="csl-bib-body">
<div class="csl-entry">Hausjell, J., Weissensteiner, J., Molitor, C., Schlangen, K., Spadiut, O., & Halbwirth, H. (2022). First purified recombinant CYP75B including transmembrane helix with unexpected high substrate specificity to (2R)-naringenin. <i>Scientific Reports</i>, <i>12</i>(1), Article 8548. https://doi.org/10.1038/s41598-022-11556-3</div>
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dc.identifier.issn
2045-2322
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dc.identifier.uri
http://hdl.handle.net/20.500.12708/148169
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dc.description.abstract
Anthochlor pigments (chalcones and aurones) play an important role in yellow flower colourization, the formation of UV-honey guides and show numerous health benefits. The B-ring hydroxylation of chalcones is performed by membrane bound cytochrome P450 enzymes. It was assumed that usual flavonoid 3'-hydroxlases (F3'Hs) are responsible for the 3,4- dihydroxy pattern of chalcones, however, we previously showed that a specialized F3'H, namely chalcone 3-hydroxylase (CH3H), is necessary for the hydroxylation of chalcones. In this study, a sequence encoding membrane bound CH3H from Dahlia variabilis was recombinantly expressed in yeast and a purification procedure was developed. The optimized purification procedure led to an overall recovery of 30% recombinant DvCH3H with a purity of more than 84%. The enzyme was biochemically characterized with regard to its kinetic parameters on various substrates, including racemic naringenin, as well as its enantiomers (2S)-, and (2R)-naringenin, apigenin and kaempferol. We report for the first time the characterization of a purified Cytochrome P450 enzyme from the flavonoid biosynthesis pathway, including the transmembrane helix. Further, we show for the first time that recombinant DvCH3H displays a higher affinity for (2R)-naringenin than for (2S)-naringenin, although (2R)-flavanones are not naturally formed by chalcone isomerase.
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dc.description.sponsorship
FWF Fonds zur Förderung der wissenschaftlichen Forschung (FWF)
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dc.language.iso
en
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dc.publisher
Nature Publishing
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dc.relation.ispartof
Scientific Reports
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dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
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dc.subject
Cytochrome P-450 Enzyme System
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dc.subject
Flavonoids
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dc.subject
Plant Proteins
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dc.subject
Substrate Specificity
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dc.subject
Chalcone
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dc.subject
Chalcones
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dc.subject
Flavanones
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dc.title
First purified recombinant CYP75B including transmembrane helix with unexpected high substrate specificity to (2R)-naringenin