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Thraustochytrids are marine protists found throughout the world in estuarine and marine habitats. These microorganisms have attracted interest, because their lipids contain different long chain polyunsaturated fatty acids (PUFAs). Those able to produce docosahexaenoic acid (C22:6 n-3, DHA) are the most studied because of the physiological importance of this PUFA in human beings. Their heterotrophic cultivation offers several challenges since biomass, lipid content, and fatty acid profile are dependent on growth conditions. In this work the effects of C source and its concentration on DHA production by different thraustochytrid strains are reviewed. Results obtained by different investigators on the use of alternative and low cost nutrient sources for production of DHA by thraustochytrids are also presented.

Production of docosahexasaenoic acid (DHA) by a native Labyrinthulomycetes strain, Thraustochytriidae sp. TN5, whose growth characteristics present some differences to related strains, was scaled from shaken flask to a laboratory fermentor. The effect of the growth medium composition and growth conditions for (i) biomass production, (ii) lipid content of the biomass, and (iii) DHA content in the lipids was determined. Taguchi's design of experiments was used to study the influence of two discrete -carbon and nitrogen sources -and six continuous -concentrations of the carbon and nitrogen sources, yeast extract and artificial seawater, incubation temperature, and time-factors on the three mentioned variables. In the flask experiments the lipidic content in the biomass (25.2% w/w) and DHA concentration (0.48 g/L) were the highest at the following conditions: maltose 20 g/L, sodium glutamate 2.4 g/L, yeast extract 6 g/L, 72 h, 258C, and artificial seawater 30% v/v; under these conditions biomass concentration was 5.1 g/L. Fed-batch allowed to increase biomass concentration to 14 g/L. The lipidic fraction of Thraustochytriidae sp. TN5 biomass in the repeated batches was found between 16.2 and 34.8% w/w. Under this growth condition lipids and DHA productivities were 50 and 23 mg/(L h), respectively.Practical applications: Consumption of long chain-polyunsaturated fatty acids (LC-PUFA) belonging to the omega-3 family such as DHA has several positive effects on human and animal health. However, natural sources are restricted to cold-water fish and their oils. Marine protists are also good candidates for the production of microbial DHA. For evaluating the potential of new strains the effect of the growth medium composition, growth conditions, and cultivation mode on DHA productivity has to be determined. In this work the production of DHA by a native Labyrinthulomycetes strain was scaled from shaken flask to a laboratory fermentor with the aid of Taguchi's methodology, process in which biomass concentration, lipid content in the biomass, and DHA content in the lipids were increased.Abbreviations: C, concentration of the carbon source; CS, carbon source; DHA, docosahexasaenoic acid; DO, dissolved oxygen; DOE, design of experiments; FAME, fatty acid methyl ester; LC-PUFA, long chainpolyunsaturated fatty acids; N, concentration of the nitrogen source;

Production of biomass and lipids in batch cultures of the Antarctic thraustochytrid Oblongichytrium sp. RT2316‐13, is reported. The microorganism proved capable of producing nearly 67% docosahexaenoic acid (DHA) and 15% eicosapentaenoic acid (EPA) in its total lipid fraction. Biomass with a maximum total lipid content of 33.5% (wt/wt) could be produced at 15°C in batch culture using a medium containing glucose (20 g/L), yeast extract (10.5 g/L), and other minor components. A lower culture temperature (5°C) reduced biomass and lipid productivities compared to culture at 15°C, but enhanced the DHA and EPA content of the lipids by 6.4‐ and 3.3‐fold, respectively. Both a simple minimally structured mathematical model and a more complex genome‐scale metabolic model (GEM) allowed the fermentation profiles in batch cultures to be satisfactorily simulated, but the GEM provided much greater insight in the biochemical and physiological phenomena underlying the observed behavior. Unlike the simpler model, the GEM could be interrogated for the possible effects of various external factors such as oxygen supply, on the expected outcomes. In silico predictions of oxygen effects were consistent with literature observations for DHA producing thraustochytrids.

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Eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and carotenoids are needed as human dietary supplements and are essential components in commercial feeds for the production of aquacultured seafood. Microorganisms such as thraustochytrids are potential natural sources of these compounds. This research reports on the lipid and carotenoid production capacity of thraustochytrids that were isolated from coastal waters of Antarctica. Of the 22 isolates, 21 produced lipids containing EPA+DHA, and the amount of these fatty acids exceeded 20% of the total fatty acids in 12 isolates. Ten isolates were shown to produce carotenoids (27.4–63.9 μg/g dry biomass). The isolate RT2316-16, identified as Thraustochytrium sp., was the best producer of biomass (7.2 g/L in five days) rich in carotenoids (63.9 μg/g) and, therefore, became the focus of this investigation. The main carotenoids in RT2316-16 were β-carotene and canthaxanthin. The content of EPA+DHA in the total lipids (34 ± 3% w/w in dry biomass) depended on the stage of growth of RT2316-16. Lipid and carotenoid content of the biomass and its concentration could be enhanced by modifying the composition of the culture medium. The estimated genome size of RT2316-16 was 44 Mb. Of the 5656 genes predicted from the genome, 4559 were annotated. These included genes of most of the enzymes in the elongation and desaturation pathway of synthesis of ω-3 polyunsaturated fatty acids. Carotenoid precursors in RT2316-16 were synthesized through the mevalonate pathway. A β-carotene synthase gene, with a different domain organization compared to the gene in other thraustochytrids, explained the carotenoid profile of RT2316-16.