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Merck

The invertebrate Caenorhabditis elegans biosynthesizes ascorbate.

Archives of biochemistry and biophysics (2015-02-11)
Alexander N Patananan, Lauren M Budenholzer, Maria E Pedraza, Eric R Torres, Lital N Adler, Steven G Clarke
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l-Ascorbate, commonly known as vitamin C, serves as an antioxidant and cofactor essential for many biological processes. Distinct ascorbate biosynthetic pathways have been established for animals and plants, but little is known about the presence or synthesis of this molecule in invertebrate species. We have investigated ascorbate metabolism in the nematode Caenorhabditis elegans, where this molecule would be expected to play roles in oxidative stress resistance and as cofactor in collagen and neurotransmitter synthesis. Using high-performance liquid chromatography and gas-chromatography mass spectrometry, we determined that ascorbate is present at low amounts in the egg stage, L1 larvae, and mixed animal populations, with the egg stage containing the highest concentrations. Incubating C. elegans with precursor molecules necessary for ascorbate synthesis in plants and animals did not significantly alter ascorbate levels. Furthermore, bioinformatic analyses did not support the presence in C. elegans of either the plant or the animal biosynthetic pathway. However, we observed the complete (13)C-labeling of ascorbate when C. elegans was grown with (13)C-labeled Escherichia coli as a food source. These results support the hypothesis that ascorbate biosynthesis in invertebrates may proceed by a novel pathway and lay the foundation for a broader understanding of its biological role.

MATERIALS
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Supelco
Dehydrated Alcohol, Pharmaceutical Secondary Standard; Certified Reference Material
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Methanol, suitable for NMR (reference standard)
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Methyl viologen dichloride hydrate, 98%
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D-Glucose-12C6, 16O6, 99.9 atom % 16O, 99.9 atom % 12C
Supelco
Ethanol solution, certified reference material, 2000 μg/mL in methanol
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D-(+)-Glucose, analytical standard
Sigma-Aldrich
Tris(2-carboxyethyl)phosphine hydrochloride, BioUltra, ≥98% (NMR)
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Butylated Hydroxytoluene, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Butylated hydroxytoluene, United States Pharmacopeia (USP) Reference Standard
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D-(+)-Glucose, BioUltra, anhydrous, โ‰ฅ99.5% (sum of enantiomers, HPLC)
Supelco
Methanol, analytical standard
Supelco
N,O-Bis(trimethylsilyl)trifluoroacetamide with trimethylchlorosilane, derivatization grade (GC derivatization), LiChropurโ„ข, contains 1% TMCS, 99% (excluding TMCS)
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Butylated hydroxytoluene, ≥99%, FCC, FG
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Chlorotrimethylsilane, Wacker Chemie AG, ≥99.0% (GC)
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D-(+)-Glucose, tested according to Ph. Eur.
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Chlorotrimethylsilane, puriss., ≥99.0% (GC)
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Chlorotrimethylsilane, ≥98.0% (GC)
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Chlorotrimethylsilane, purified by redistillation, ≥99%
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Citrate Concentrated Solution, BioUltra, Molecular Biology, 1 M in H2O
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D-(+)-Galactose, meets analytical specification of Ph. Eur., BP
Supelco
N,O-Bis(trimethylsilyl)trifluoroacetamide with trimethylchlorosilane, derivatization grade (GC derivatization), LiChropurโ„ข, contains 10% TMCS, 98% (excluding TMCS)
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D-Glucuronic acid, โ‰ฅ98% (GC)
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D-(+)-Galactose, BioXtra, โ‰ฅ99% (HPLC)
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D-(+)-Galactose, โ‰ฅ99% (HPLC)
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D-(+)-Glucose, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, โ‰ฅ99.5%
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D-(+)-Galactose, โ‰ฅ98% (HPLC)
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D-(+)-Glucose, Hybri-Maxโ„ข, powder, BioReagent, suitable for hybridoma