Alternativer Identifier:
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Verwandter Identifier:
(Is Supplement To) 10.1111/tpj.15847 - DOI
Ersteller/in:
Jäckel, Luise [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]

Stellmach, Hagen https://orcid.org/0000-0002-7206-974X [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]

Klauß, Ulrike [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]

Matschi, Susanne https://orcid.org/0000-0001-6233-2519 [Leibniz Institute of Plant Biochemistry, Dept. Biochemistry of Plant Interactions, Weinberg 3, 06120 Halle (Saale), Germany]

Hause, Gerd https://orcid.org/0000-0003-0242-3923 [Martin-Luther-University Halle-Wittenberg; Biocenter, Electron Microscopy Lab, Weinbergweg 22, 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]
Beitragende:
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Titel:
Supplement: The terminal enzymatic step in piperine biosynthesis is co-localized with the product piperine in specialized cells of black pepper (Piper nigrum L.)
Weitere Titel:
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Beschreibung:
(Abstract) Piperine (1-piperoyl piperidine) is responsible for the pungent perception of dried black pepper (Piper nigrum) fruits and essentially contributes to the aromatic properties of this spice in combination with ablend of terpenoids. The final step in piperine biosynthesis involves piperine synthase (PS), which catalyzes the reaction of piperoyl CoA and piperidine to the biologically active and pungent amide. Nevertheless, experimental data on the cellular localization of piperine and the complete biosynthetic pathway are missing. Not only co-localization of enzymes and products, but also potential transport of piperamides to the sink organs is a possible alternative. This work, which includes purification of the native enzyme, immunolocalization, laser microdissection, fluorescence microscopy, and electron microscopy combined with liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), provides experimental evidence that piperine and PS are co-localized in specialized cells of the black pepper fruit perisperm. PS accumulates during early stages of fruit development and its level declines before the fruits are fully mature. The product piperine is co-localized to PS and can be monitored at the cellular level by its strong bluish fluorescence. Rising piperine levels during fruit maturation are consistent with the increasing numbers of fluorescent cells within the perisperm. Signal intensities of individual laser-dissected cells when monitored by LC-ESI-MS/MS indicate molar concentrations of this alkaloid. Significant levels of piperine and additional piperamides were also detected in cells distributed in the cortex of black pepper roots. In summary, the data provide comprehensive experimental evidence of and insights into cell-specific biosynthesis and storage of piperidine alkaloids, specific and characteristic for the Piperaceae. By a combination of fluorescence microscopy and LC-MS/MS analysis we localized the major piperidine alkaloids to specific cells of the fruit perisperm and the root cortex. Immunolocalization of native piperine and piperamide synthases shows that enzymes are co-localized with high concentrations of products in these idioblasts.
Schlagworte:
Piper nigrum, piperine, specialized metabolism, alkaloid, enzyme purification, fluorescence microscopy, laser microdissection
Zugehörige Informationen:
(human readable resource description) arw files (chromatogram raw data), czi Files (microscopy raw data), csv and Excel files (xlsx), raw files (protein LC-MS/MS) tiff and jpg metadata by imaging machines
(InChI) InChI=1S/C17H19NO3/c19-17(18-10-4-1-5-11-18)7-3-2-6-14-8-9-15-16(12-14)21-13-20-15/h2-3,6-9,12H,1,4-5,10-11,13H2/b6-2+,7-3+
(InChIKey) MXXWOMGUGJBKIW-YPCIICBESA-N
(RInChI) RInChI=1.00.1S/C17H19NO3/c19-17(18-10-4-1-5-11-18)7-3-2-6-14-8-9-15-16(12-14)21-13-20-15/h2-3,6-9,12H,1,4-5,10-11,13H2/b6-2+,7-3+!C21H36N7O16P3S/c1-21(2,16(31)19(32)24-4-3-12(29)23-5-6-48)8-41-47(38,39)44-46(36,37)40-7-11-15(43-45(33,34)35)14(30)20(42-11)28-10-27-13-17(22)25-9-26-18(13)28/h9-11,14-16,20,30-31,48H,3-8H2,1-2H3,(H,23,29)(H,24,32)(H,36,37)(H,38,39)(H2,22,25,26)(H2,33,34,35)/t11-,14-,15-,16+,20-/m1/s1<>C33H44N7O19P3S/c1-33(2,28(44)31(45)36-10-9-23(41)35-11-12-63-24(42)6-4-3-5-19-7-8-20-21(13-19)54-18-53-20)15-56-62(51,52)59-61(49,50)55-14-22-27(58-60(46,47)48)26(43)32(57-22)40-17-39-25-29(34)37-16-38-30(25)40/h3-8,13,16-17,22,26-28,32,43-44H,9-12,14-15,18H2,1-2H3,(H,35,41)(H,36,45)(H,49,50)(H,51,52)(H2,34,37,38)(H2,46,47,48)/b5-3+,6-4+/t22-,26-,27-,28+,32-/m1/s1!C5H11N/c1-2-4-6-5-3-1/h6H,1-5H2/d-
(Long-RInChIKey) Long-RInChIKey=SA-BUHFF-MXXWOMGUGJBKIW-YPCIICBESA-N-RGJOEKWQDUBAIZ-IBOSZNHHSA-N--GEVZCNXLEOONCV-TZKXQVKESA-N-NQRYJNQNLNOLGT-UHFFFAOYSA-N
(Short-RInChIKey) Short-RInChIKey=SA-BUHFF-QWBPSOYVMQ-JSBKXZXFNT-UHFFFADPSC-NKALM-NURCX-NUHFF-ZZZ
(Web-RInChIKey) Web-RInChIKey=RNGMSRAOERGJTOQUV-NQHBIAWVHCLSFSA
Sprache:
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Herausgeber/in:
Vogt, Thomas
Erstellungsjahr:
Fachgebiet:
Biochemistry
Objekttyp:
(Dataset) Supplemental data
Datenquelle:
(Instrument) Waters e2695 chromatography work station equipped with a photodiode array detector (PDA) and a QDA-mass detector (Waters)
(Instrument) laser scanning microscope LSM 780 - Carl Zeiss
(Instrument) cryomicrotome 1950 - Carl Zeiss
(Instrument) transmission electron microscope EM900 - Carl Zeiss
(Instrument) Äkta Explorer FPLC - Cytiva, Freiburg
(Instrument) QExactive Plus mass spectrometer - Thermo Fisher Scientific
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Carl Zeiss Zen - 2012
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Waters Empower - 3
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Software:
Unicorn - 5.20
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Software für Datenerhebung
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Mascot - v2.7.0
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Software für Datenerhebung
Software:
Thermo Fisher Proteome Discoverer - v2.1
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Software für Datenbearbeitung
Software:
Microsoft Office - 2013
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Software für Datenbearbeitung
Software:
Systat SigmaPlot - 14.0
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Software für Datenbearbeitung
Software:
Serif Affinity Graphics - unknown
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Datenverarbeitung:
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Erscheinungsjahr:
Lizenz:
Rechteinhaber/in:
Leibniz-Institut für Pflanzenbiochemie (IPB), Halle (Saale), Germany
Förderung:
Deutsche Forschungsgemeinschaft (DFG) - (VO719/15-1/2)
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Status:
Publiziert
Eingestellt von:
1670fa88949cc897f41b44d58d08c11a
Erstellt am:
Archivierungsdatum:
2022-06-14
Archivgröße:
5,7 GB
Archiversteller:
1670fa88949cc897f41b44d58d08c11a
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ab9b6fdecce1ae51f52ce931b81c4db2 (MD5)
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