Journal
TOXICON
Volume 92, Issue -, Pages 1-5Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.toxicon.2014.09.003
Keywords
Domoic acid (DA); Amnesic shellfish poisoning (ASP); Red alga; Chondria am-rata; Laboratory culture; Modified PBS medium
Categories
Funding
- Ministry of Education, Science and Culture, Japan [22580379, 23580282]
- Grants-in-Aid for Scientific Research [23580282, 22580379] Funding Source: KAKEN
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To clarify the production mechanisms and biologic functions of domoic acid (DA) by the red alga Chondria armata, we established a laboratory culture of C armata. The alga grew better in modified PBS medium (mPES) without trace metals or manganese than in unmodified mPES (seawater nitrate, phosphate, iron, trace metals, vitamins, and 2-[4-2-hydroxyethyl)-1-piperazinyl]-ethanesulfonic acid), suggesting that C. armata is especially hypersensitive to the toxicity of excessive manganese. C. armata cultured in N.P.Fe medium (seawater nitrate, phosphate, and iron) grew best (mean growth rate 828.4%) at a relative nutrient concentration of 50%. Liquid chromatography-mass spectrometry analysis of the algal extracts revealed that the DA content of the cultured explants (2273-3308 ppm) was 4-5 fold higher than that of wild specimens. The extract of pooled explants (60 g) was purified by activated charcoal treatment and several types of column chromatography to afford ca. 10 mg DA. The H-1-nuclear magnetic resonance spectrum of the preparation was indistinguishable from the previously reported spectrum of DA, indicating that C. armata itself has an ability to produce DA. (C) 2014 Elsevier Ltd. All rights reserved.
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