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

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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

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Sum total of downloads: 130




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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.
License of this version: Es gilt deutsches Urheberrecht. Das Dokument darf zum eigenen Gebrauch kostenfrei genutzt, aber nicht im Internet bereitgestellt oder an Außenstehende weitergegeben werden.
Document Type: article
Publishing status: acceptedVersion
Issue Date: 2016
Appears in Collections:Naturwissenschaftliche Fakultät

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pos. country downloads
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1 image of flag of Germany Germany 77 59.23%
2 image of flag of China China 17 13.08%
3 image of flag of India India 7 5.38%
4 image of flag of United States United States 3 2.31%
5 image of flag of Nigeria Nigeria 3 2.31%
6 image of flag of Malaysia Malaysia 3 2.31%
7 image of flag of France France 3 2.31%
8 image of flag of Belgium Belgium 3 2.31%
9 image of flag of Indonesia Indonesia 2 1.54%
10 image of flag of Canada Canada 2 1.54%
    other countries 10 7.69%

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