Heterologous Expression, Purification, and Biochemical Characterization of α-Humulene Synthase from Zingiber zerumbet Smith

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dc.identifier.uri http://dx.doi.org/10.15488/1388
dc.identifier.uri http://www.repo.uni-hannover.de/handle/123456789/1413
dc.contributor.author Alemdar, Semra
dc.contributor.author Hartwig, Steffen
dc.contributor.author Frister, Thore
dc.contributor.author König, Jan Christoph
dc.contributor.author Scheper, Thomas
dc.contributor.author Beutel, Sascha
dc.date.accessioned 2017-04-21T11:19:50Z
dc.date.available 2017-04-21T11:19:50Z
dc.date.issued 2016
dc.identifier.citation Alemdar, S.; Hartwig, S.; Frister, T.; König, J.C.; Scheper, T.; Beutel, S.: Heterologous Expression, Purification, and Biochemical Characterization of α-Humulene Synthase from Zingiber zerumbet Smith. In: Applied Biochemistry and Biotechnology 178 (2016), Nr. 3, S. 474-489. DOI: https://doi.org/10.1007/s12010-015-1888-4
dc.description.abstract The α-humulene synthase from Zingiber zerumbet Smith was expressed as a polyhistidine-tagged protein in an E. coli BL21(DE3) strain. Induction time and inductor (isopropyl-β-D-thiogalactopyranoside) concentration were optimized. The enzyme was successfully purified directly from cell lysate by NTA affinity column chromatography and careful selection of coordinated metal ion and imidazole elution conditions. Bioactivity assays were conducted with the natural substrate farnesyl diphosphate (FDP) in a two-phase system with in situ extraction of products. The conversion of FDP to α-humulene (~94.5 %) and β-caryophyllene (~5.5 %) could be monitored by gas chromatography-flame ionization detection (GC-FID). Optimal pH and temperature as well as kinetic parameters KM and kcat were determined using a discontinuous kinetic assay. The final publication is available at Springer via http://dx.doi.org/10.1007/s12010-015-1888-4. eng
dc.description.sponsorship EFRE/ZW-8-80130940
dc.language.iso eng
dc.publisher New York City, NY : Springer Humana Press Inc.
dc.relation.ispartofseries Applied Biochemistry and Biotechnology 178 (2016), Nr. 3
dc.rights Es gilt deutsches Urheberrecht. Das Dokument darf zum eigenen Gebrauch kostenfrei genutzt, aber nicht im Internet bereitgestellt oder an Außenstehende weitergegeben werden.
dc.subject Enzyme activity eng
dc.subject Humulene eng
dc.subject Purification eng
dc.subject Recombinant expression eng
dc.subject Sesquiterpene eng
dc.subject Terpene synthase eng
dc.subject Assays eng
dc.subject Chromatography eng
dc.subject Enzymes eng
dc.subject Escherichia coli eng
dc.subject Gas chromatography eng
dc.subject Ionization of gases eng
dc.subject Metal ions eng
dc.subject Metals eng
dc.subject Biochemical characterization eng
dc.subject Farnesyl diphosphate eng
dc.subject Gas chromatography-flame ionization detections eng
dc.subject Heterologous expression eng
dc.subject Humulene eng
dc.subject Recombinant expression eng
dc.subject Sesquiterpenes eng
dc.subject Synthases eng
dc.subject Column chromatography eng
dc.subject.ddc 570 | Biowissenschaften, Biologie ger
dc.subject.ddc 540 | Chemie ger
dc.title Heterologous Expression, Purification, and Biochemical Characterization of α-Humulene Synthase from Zingiber zerumbet Smith
dc.type article
dc.type Text
dc.relation.essn 0273-2289
dc.relation.doi 10.1007/s12010-015-1888-4
dc.bibliographicCitation.issue 3
dc.bibliographicCitation.volume 178
dc.bibliographicCitation.firstPage 474
dc.bibliographicCitation.lastPage 489
dc.description.version acceptedVersion
tib.accessRights frei zug�nglich


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