Catalytical Specificity, Reaction Mechanisms, and Conformational Changes during Catalysis of the Recombinant SUMO (+)-Zizaene Synthase from Chrysopogon zizanioides

Show simple item record

dc.identifier.uri http://dx.doi.org/10.15488/4713
dc.identifier.uri https://www.repo.uni-hannover.de/handle/123456789/4755
dc.contributor.author Aguilar Cascante, Francisco ger
dc.contributor.author Hartwig, Steffen ger
dc.contributor.author Scheper, Thomas ger
dc.contributor.author Beutel, Sascha ger
dc.date.accessioned 2019-04-16T13:03:40Z
dc.date.available 2019-04-16T13:03:40Z
dc.date.issued 2019
dc.identifier.citation Aguilar Cascante, F.; Hartwig, S.; Scheper, T.; Beutel, S.: Catalytical Specificity, Reaction Mechanisms, and Conformational Changes during Catalysis of the Recombinant SUMO (+)-Zizaene Synthase from Chrysopogon zizanioides. In: ACS Omega 4 (2019), Nr. 4, S. 6199-6209. DOI: http://doi.org/10.1021/acsomega.9b00242 ger
dc.description.abstract Zizaene synthase (ZS) from Chrysopogon zizanioides (Poaceae) is the critical enzyme in the biosynthesis of the fragrant sesquiterpene khusimol, a major component of the vetiver essential oil used widely by the cosmetic industry. As reported previously, we heterologously and successfully ex-pressed the active ZS with a small ubiquitin-related modifier (SUMO) fusion domain. In this study, we report the optimization of reaction conditions and determination of enzyme kinetics of ZS. Moreover, we investigate the catalytic specificity and reaction mechanisms with the ubiquitous (2E,6E)-farnesyl diphosphate (FDP) and with C10 and C15 prenyl diphosphate isomers. Catalytic promiscuity occurs with monoterpene substrates generating eight products that comprise acyclic, cyclic, and hydroxylated monoterpenes. In contrast, ZS is a high-fidelity terpene cyclase when used with C15 isomer substrates, yielding as major products (Z)-β-farnesene (100%) for (2E,6Z)-FDP and (+)-zizaene (81.7%), β-acoradiene (12.8%), and (E)-β-farnesene (5.5%) for (2Z,6E)-FDP. Cyclization of the ubiquitous substrate (2E,6E)-FDP demonstrates a higher catalytic specificity, whereas the reaction proceeds via the acorenyl cation that generates (+)-zizaene (91.5%) and β-acoradiene (8.5%). Furthermore, catalytic specificity with (2E,6E)-FDP was stable in reactions tested at distinct pH and temperatures, suggesting a stable and efficient closed conformation of the active site during catalysis. To understand such stability, open and closed structural conformations of ZS were modeled in silico and revealed putative residues in the active site and in the A-C and J-K surrounding loops, which could explain the high fidelity of ZS. ger
dc.language.iso eng ger
dc.publisher Washington D.C., USA : ACS Publications
dc.relation.ispartofseries ACS Omega 4 (2019), Nr. 4 ger
dc.rights ACS AuthorChoice License ger
dc.rights.uri https://pubs.acs.org/page/policy/authorchoice_termsofuse.html ger
dc.subject Sesquiterpene synthase eng
dc.subject Zizaene eng
dc.subject Catalytical Specificity eng
dc.subject Sesquiterpensynthasen ger
dc.subject Zizaen ger
dc.subject Katalytische Spezifität ger
dc.subject.ddc 500 | Naturwissenschaften ger
dc.subject.ddc 540 | Chemie ger
dc.title Catalytical Specificity, Reaction Mechanisms, and Conformational Changes during Catalysis of the Recombinant SUMO (+)-Zizaene Synthase from Chrysopogon zizanioides eng
dc.type article ger
dc.type Text ger
dc.relation.doi 10.1021/acsomega.9b00242
dc.description.version publishedVersion ger
tib.accessRights frei zug�nglich ger
tib.accessRights frei zug�nglich


Files in this item

This item appears in the following Collection(s):

Show simple item record

 

Search the repository


Browse

My Account

Usage Statistics