Analysis of docosahexaenoic acid biosynthesis in Crypthecodinium cohnii by 13C labelling and desaturase inhibitor experiments

M.E. de Swaaf, T.C. de Rijk, P. van der Meer, G. Eggink, L. Sijtsma

Research output: Contribution to journalArticleAcademicpeer-review

43 Citations (Scopus)

Abstract

The lipids of the heterotrophic microalga Crypthecodinium cohnii contain the omega-3 polyunsaturated fatty acid (PUFA) and docosahexaenoic acid (22:6) to a level of over 30&Eth;The pathway of 22:6 synthesis in C. cohnii is unknown. The ability of C. cohnii to use 13C-labelled externally supplied precursor molecules for 22:6 biosynthesis was tested by 13C NMR analysis. Furthermore, the presence of desaturases (typical for aerobic PUFA synthesis) was studied by the addition of specific desaturase inhibitors in the growth medium. The addition of 1-13C acetate or 1-13C butyrate in the growth medium resulted in 22:6 with only the odd carbon atoms enriched. Apparently, two-carbon units were used as building blocks for 22:6 synthesis and butyrate was first split into two-carbon units prior to incorporation in 22:6. When 1-13C oleic acid was added to the growth medium, 1-13C oleic acid was incorporated into the lipids of C. cohnii but was not used as a precursor for the synthesis of 22:6. Specific desaturase inhibitors (norflurazon and propyl gallate) inhibited lipid accumulation in C. cohnii. The fatty acid profile, however, was not altered. In contrast, in the arachidonic acid-producing fungus, Mortierella alpina, these inhibitors not only decreased the lipid content but also altered the fatty acid profile. Our results can be explained by the presence of three tightly regulated separate systems for the fatty acid production by C. cohnii, namely for (1) the biosynthesis of saturated fatty acids, (2) the conversion of saturated fatty acids to monounsaturated fatty acids and (3) the de novo synthesis of 22:6 with desaturases involved.
Original languageEnglish
Pages (from-to)21-29
JournalJournal of Biotechnology
Volume103
Issue number1
DOIs
Publication statusPublished - 2003

Keywords

  • n-3 fatty-acids
  • marine microalgae
  • omega-3-fatty-acids
  • triacylglycerols

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