Alternate identifier:
(InChIKey) YKPUWZUDDOIDPM-SOFGYWHQSA-N
(InChI) InChI=1S/C18H27NO3/c1-14(2)8-6-4-5-7-9-18(21)19-13-15-10-11-16(20)17(12-15)22-3/h6,8,10-12,14,20H,4-5,7,9,13H2,1-3H3,(H,19,21)/b8-6+
(CAS/RN) 404-86-4
(InChIKey) PUDMGOSXPCMUJZ-UHFFFAOYSA-N
(InChI) InChI=1S/C8H11NO2.ClH/c1-11-8-4-6(5-9)2-3-7(8)10;/h2-4,10H,5,9H2,1H3;1H
(CAS/RN) 7149-10-2
(InChIKey) JGNCYWQFXLYWMO-HSJNEKGZSA-N
(InChI) InChI=1S/C31H52N7O17P3S/c1-19(2)9-7-5-6-8-10-22(40)59-14-13-33-21(39)11-12-34-29(43)26(42)31(3,4)16-52-58(49,50)55-57(47,48)51-15-20-25(54-56(44,45)46)24(41)30(53-20)38-18-37-23-27(32)35-17-36-28(23)38/h7,9,17-20,24-26,30,41-42H,5-6,8,10-16H2,1-4H3,(H,33,39)(H,34,43)(H,47,48)(H,49,50)(H2,32,35,36)(H2,44,45,46)/t20-,24-,25-,26+,30-/m1/s1
Related identifier:
Creator/Author:
Milde, Raika https://orcid.org/0000-0003-0630-8105 [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]

Schnabel, Arianne https://orcid.org/0000-0002-2172-2905 [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]

Ditfe, Toni https://orcid.org/0000-0001-6360-6673 [Leibniz Institute of Plant Biochemistry, Dept. of Bioorganic Chemistry, Weinberg 3, 06120 Halle (Saale), Germany]

Hoehenwarter, Wolfgang https://orcid.org/0000-0002-7669-7524 [Leibniz Institute of Plant Biochemistry, Dept. of Biochemistry of Plant Interactions, Weinberg 3, 06120 Halle (Saale), Germany]

Proksch, Carsten https://orcid.org/0000-0003-4948-8281 [Leibniz Institute of Plant Biochemistry, Dept. of Biochemistry of Plant Interactions, Weinberg 3, 06120 Halle (Saale), Germany]

Westermann, Bernhard https://orcid.org/0000-0002-6228-5991 [Leibniz Institute of Plant Biochemistry, Dept. of Bioorganic Chemistry, Weinberg 3, 06120 Halle (Saale), Germany]

Vogt, Thomas https://orcid.org/0000-0001-6155-3360 [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]
Contributors:
-
Title:
Supporting information: Chemical synthesis of trans 8-methyl-6-nonenoyl-CoA and functional expression unravel capsaicin synthase activity encoded by the Pun1 locus
Additional titles:
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Description:
(Abstract) Capsaicin, produced by diverse Capsicum species is among the world’s most popular spices and of considerable pharmaceutical relevance. Although the capsaicinoid biosynthetic pathway has been investigated for decades, several biosynthetic steps have remained partly hypothetical. Genetic evidence suggested that the decisive capsaicin synthase is encoded by the Pun1 locus. Yet, genetic evidence of the Pun1 locus was never corroborated by functionally active capsaicin synthase that presumably catalyzes amide bond formation between trans 8-methyl-6-nonenoyl-CoA derived from branched chain amino acid biosynthesis and vanilloylamine derived from the phenylpropanoid pathway. In this report, we demonstrate enzymatic activity of a recombinant capsaicin synthase encoded by Pun1, functionally expressed in E. coli, and provide information on substrate specificity and catalytic properties. Recombinant capsaicin synthase is specific for selected aliphatic CoA-esters and highly specific for vanilloylamine. Partly purified from E. coli, the recombinant active enzyme is a monomeric protein of 51 kDa that is independent of additional co-factors or associated proteins, as previously proposed. These data can now be used to design capsaicin synthase variants with different properties and alternative substrate preferences.
Keywords:
Capsaicin
Capsicum
Pun1
acyltransferase
BAHD
CoA-ester
enzyme activity
substrate specificity
Related information:
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Language:
-
Publishers:
Vogt, Thomas
Production year:
Subject areas:
Biochemistry
Resource type:
(Dataset) chromatogram data (arw files), protein LC-MS/MS, images (tiff, jpeg), tabular data (csv, xlsx)
Data source:
(Instrument) Varian Mercury 400 NMR spectrometer (Agilent, Waldbronn, Germany)
(Instrument) AB Sciex API 3200, Q-TRAP mass spectrometer (AB Sciex , Darmstadt, Germany)
(Instrument) Waters e2695 chromatography work station equipped with a photodiode array detector (PDA) and a QDA-mass detector (Waters, Eschborn, Germany)
(Instrument) Äkta Explorer FPLC (Cytiva, Freiburg, Germany) Instrument
(Instrument) QExactive Plus mass spectrometer (Thermo Fisher Scientific, Dreieich, Germany)
(Organism) Capsicum
(Organism) Escherichia coli
Software used:
Resource production
Software:
VnmrJ (Agilent) - 4.2 Revision A
Alternative software:
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Software used:
Resource production
Software:
AB Sciex Analyst - 1.6.2
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Resource processing
Software:
MestReNova - 14.0.1
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Resource processing
Software:
ProteoWizard - 3.0.2
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Resource production
Software:
Waters Empower (Waters) - 3
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Software used:
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Software:
Carl Zeiss Zen (Carl Zeiss AG) - 2012
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Resource processing
Software:
Unicorn (Amersham Biosciences, now Cytiva ) - 5.0
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Software used:
Resource production
Software:
Mascot (Matrix Science) - v2.7.0
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Software used:
Resource production
Software:
Thermo Fisher Scientific Proteom Discoverer - v2.1
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Software used:
Resource processing
Software:
Microsoft Office - 2013
Alternative software:
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Software used:
Resource processing
Software:
Systat SigmaPlot (Systat) - 14.0
Alternative software:
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Data processing:
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Publication year:
Rights holders:
Leibniz Institute of Plant Biochemistry (IPB), Dept. Cell and Metabolic Biology and Dept. Bioorganic Chemistry, Weinberg 3, 06120 Halle (Saale), Germany.
Funding:
-
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2022-10-19
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