Abstract
Thirteen different14C-and3H-labelled epicuticular wax precursors have been synthesized and their structure determined by gas chromatography-mass spectrometry analyses. The biosyntheses of β-diketones and alkan-2-ol containing esters were studied by incorporating these intermediates into tissue slices of barley spikes whose awns had been removed. A differential labelling pattern of the alkan-2-ol esters and the β-diketones was observed after feeding three selected mutants blocked in different steps catalyzed by a multifunctional enzyme encoded for by thecer-cqu gene. Incer-u 69 tissue slices (9,10-3H)-3-oxopalmitoyl-CoA was incorporated into both the esterified alkan-2-ols and the β-diketones. Only the former wax component was synthesized by the mutantscer-c 36, and-q 42. When C14 and C16 fatty acyl chains were fed to the tissues slices, those ofcer-u 69 and-c 36 readily labelled the esterified alkan-2-ols, whereas those ofcer-q 42 were totally inactive. In all three mutants (2-14C)-pentadecan-2-one, (10,11-3H)-heptadecan-2-one and (2-3H)-pentadecan-2-ol exclusively labelled the alkan-2-ol moieties of the specified esters. (9,10-3H)-l-3-hydroxypalmitoyl-CoA and (3-14C)-labelleddl-3-hydroxy fatty acids having 14, 16 and 18 carbon atoms were incorporated with a very low efficiency into the β-diketones and the esterified alkan-2-ols. (9,10-3H)-3-oxopalmitoyl-CoA is the primer for the enzyme system known as β-ketoacyl elongase which forms the C29 (nonacosan-14,16-dione), C31 (hentriacontan-14, 16-dione) and C33 (tritriacontan-16,18-dione) β-diketones. After protection of the β-dicarbonyl group, 7 or 8 C2 units are added before the presumed decarboxylation to yield the complete β-diketone carbon chain. The alkan-2-ol esters arise from the 3-oxoacyl-CoA derivative by an initial decarboxylation to form a methyl ketone, followed by a reduction to an alkan-2-ol. The latter is then esterified with a fatty acid to form the alkan-2-ol containing esters. The three steps involved in the alkan-2-ol ester synthesis are accomplished by the coordinated action of a decarboxylase, reductase and ester synthetase.
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Abbreviations
- CoA:
-
coenzyme A
- GC:
-
gas chromatography
- GC-MS:
-
gas chromatography-mass spectrometry
- HPLC:
-
high performance liquid chromatography
- TLC:
-
thin layer chromatography
- TMS:
-
trimethylsilyl
- Tris:
-
tris(hydroxymethyl)-aminomethane
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Mikkelsen, J.D. Biosynthesis of esterified alkan-2-ols and β-diketones in barley spike epicuticular wax: Synthesis of radioactive intermediates. Carlsberg Res. Commun. 49, 391–416 (1984). https://doi.org/10.1007/BF02907782
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DOI: https://doi.org/10.1007/BF02907782
Keywords
- High performance liquid chromatography
- radio-gas chromatography
- mass spectrometry
- β-diol trimethylsilyl ethers
- (9,10-3H)-3-oxopalmitoyl-CoA, (1-14C)-3-oxopalmitoyl-CoA
- (9,10-3H)-L-3-hydroxypalmitoyl-CoA, (2-14C)-pentadecan-2-one
- (10,11-3H)-heptadecan-2-one
- (2-3H)-pentadecan-2-ol
- (3-14C)-3-oxopalmitic acid methyl ester
- (3-14C)-DL-3-hydroxypalmitic acid
- (1-14C)-10,12-dioxoeicosanoic acid
- reductase
- decarboxylase
- ester synthetase
- eceriferum