Details
| Original language | English |
|---|---|
| Pages (from-to) | 654-658 |
| Number of pages | 5 |
| Journal | Engineering in life sciences |
| Volume | 18 |
| Issue number | 9 |
| Early online date | 29 May 2018 |
| Publication status | Published - 3 Sept 2018 |
Abstract
Terpene synthases are the key enzymes in terpene biosynthesis that provide a structurally complex and highly diverse product spectrum. A suitable and reliable analytical assay is indispensable to measure terpene synthase activity accurately and precisely. In this study, a malachite green assay (MG) was adapted to rapidly assay terpene synthase activity and was validated in comparison to an already established gas chromatography assay. A linear correlation between both assays was observed. Kinetic properties for the previously described sesquiterpene synthase α-humulene synthase (HUM) from Zingiber zerumbet Smith were investigated for the bioconversion of the monoterpene precursors geranyl pyrophosphate (2E-GPP) and neryl pyrophosphate (2Z-NPP) as well as for the sesquiterpene precursor farnesyl pyrophosphate (2E,6E-FPP). Also, gas chromatography mass spectrometry (GS-MS) was carried out to identify the products of the bioconversion of (2E)-GPP and (2Z)-NPP.
Keywords
- Activity assay, Malachite green, Substrate specificity, Terpene synthase, α-Humulene synthase
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Environmental Science(all)
- Environmental Engineering
- Chemical Engineering(all)
- Bioengineering
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In: Engineering in life sciences, Vol. 18, No. 9, 03.09.2018, p. 654-658.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Substrate specificity of α-humulene synthase from Zingiber zerumbet Smith and determination of kinetic constants by a spectrophotometric assay
AU - Alemdar, Semra
AU - König, Jan Christoph
AU - Seidel, Katja
AU - Kirschning, Andreas
AU - Scheper, Thomas
AU - Beutel, Sascha
N1 - Funding information: This study was funded by the European Regional Development Fund (EFRE): Innovation Network “Refinement of plant resources” (ZW-8-80130940). Special thanks to Daniel Sandner and Dr. Ulrich Krings (both Institute of Food Chemistry, Leibniz Universität Hannover) for performing GS-MS measurements.
PY - 2018/9/3
Y1 - 2018/9/3
N2 - Terpene synthases are the key enzymes in terpene biosynthesis that provide a structurally complex and highly diverse product spectrum. A suitable and reliable analytical assay is indispensable to measure terpene synthase activity accurately and precisely. In this study, a malachite green assay (MG) was adapted to rapidly assay terpene synthase activity and was validated in comparison to an already established gas chromatography assay. A linear correlation between both assays was observed. Kinetic properties for the previously described sesquiterpene synthase α-humulene synthase (HUM) from Zingiber zerumbet Smith were investigated for the bioconversion of the monoterpene precursors geranyl pyrophosphate (2E-GPP) and neryl pyrophosphate (2Z-NPP) as well as for the sesquiterpene precursor farnesyl pyrophosphate (2E,6E-FPP). Also, gas chromatography mass spectrometry (GS-MS) was carried out to identify the products of the bioconversion of (2E)-GPP and (2Z)-NPP.
AB - Terpene synthases are the key enzymes in terpene biosynthesis that provide a structurally complex and highly diverse product spectrum. A suitable and reliable analytical assay is indispensable to measure terpene synthase activity accurately and precisely. In this study, a malachite green assay (MG) was adapted to rapidly assay terpene synthase activity and was validated in comparison to an already established gas chromatography assay. A linear correlation between both assays was observed. Kinetic properties for the previously described sesquiterpene synthase α-humulene synthase (HUM) from Zingiber zerumbet Smith were investigated for the bioconversion of the monoterpene precursors geranyl pyrophosphate (2E-GPP) and neryl pyrophosphate (2Z-NPP) as well as for the sesquiterpene precursor farnesyl pyrophosphate (2E,6E-FPP). Also, gas chromatography mass spectrometry (GS-MS) was carried out to identify the products of the bioconversion of (2E)-GPP and (2Z)-NPP.
KW - Activity assay
KW - Malachite green
KW - Substrate specificity
KW - Terpene synthase
KW - α-Humulene synthase
UR - http://www.scopus.com/inward/record.url?scp=85050392314&partnerID=8YFLogxK
U2 - 10.1002/elsc.201800019
DO - 10.1002/elsc.201800019
M3 - Article
C2 - 32624945
AN - SCOPUS:85050392314
VL - 18
SP - 654
EP - 658
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 9
ER -