LC-PUFA-Enriched Oil Production by Microalgae: Accumulation of Lipid and Triacylglycerols Containing n-3 LC-PUFA Is Triggered by Nitrogen Limitation and Inorganic Carbon Availability in the Marine Haptophyte Pavlova lutheri
Abstract
:1. Introduction
2. Results and Discussion
2.1. Bicarbonate Addition Promotes Growth, Nitrate Uptake and Lipid Production during Batch Cultivation of P. lutheri
2.2. Combined Effect of Nitrogen Limitation and Bicarbonate Addition on Total Fatty Acid Composition and n-3 LC-PUFA Production
Bicarbonate (mM) | ||||||
---|---|---|---|---|---|---|
Before N-Limitation | After N-Limitation | |||||
Fatty Acids (% TFA) | 2 | 9 | 18 | 2 | 9 | 18 |
Saturated fatty acids | ||||||
14:0 | 17.3 ± 0.7 | 15.2 ± 0.7 | 15.5 ± 2.8 | 14.3 ± 0.3 | 13.8 ± 0.3 | 14.7 ± 1.3 |
16:0 | 18.5 ± 1.1 | 19.7 ± 0.3 | 19.0 ± 2.0 | 26.2 ± 0.1 | 27.4 ± 0.8 | 29.9 ± 1.9 |
18:0 | 2.1 ± 1.8 | 1.9 ± 0.2 | 2.7 ± 2.5 | 0.6 ± 0.0 | 1.5 ± 0.3 | 0.7 ± 0.2 |
Sum of SFA | 37.8 ± 0.4 | 36.9 ± 1.0 | 37.2 ± 7.2 | 41.1 ± 0.2 | 42.6 ± 0.2 | 45.3 ± 0.6 |
Monounsaturated fatty acids | ||||||
16:1 n-7 | 24.6 ± 1.0 | 20.2 ± 1.3 | 17.9 ± 0.5 | 30.3 ± 0.1 | 29.4 ± 0.6 | 29.3 ± 1.6 |
18:1 n-7 | 1.3 ± 0.2 | 1.0 ± 0.1 | 1.3 ± 0.1 | 1.5 ± 0.1 | 2.2 ± 0.2 | 1.4 ± 0.1 |
18:1 n-9 | 2.0 ± 0.1 | 2.6 ± 0.6 | 2.0 ± 0.1 | 2.1 ± 0.1 | 2.9 ± 0.2 | 1.5 ± 0.2 |
Sum of MUFA | 27.8 ± 0.9 | 23.9 ± 1.4 | 21.2 ± 0.3 | 33.9 ± 0.3 | 34.4 ± 0.8 | 32.2 ± 1.4 |
Polyunsaturated fatty acids | ||||||
18:2 n-6 | 0.8 ± 0.1 | 1.2 ± 0.2 | 1.4 ± 0.3 | 1.6 ± 0.1 | 2.0 ± 0.1 | 1.9 ± 0.2 |
18:3 n-6 | tr. | 0.6 ± 0.1 | tr. | tr. | 0.3 ± 0.0 | tr. |
18:3 n-3 | 1.1 ± 0.3 | 1.1 ± 0.2 | 0.4 ± 0.1 | 0.7 ± 0.0 | 0.8 ± 0.1 | 0.6 ± 0.0 |
18:4 n-3 | 4.0 ± 0.5 | 5.7 ± 0.1 | 7.2 ± 1.4 | 2.0 ± 0.0 | 2.0 ± 0.2 | 2.6 ± 0.1 |
20:4 n-6 | 0.8 ± 0.1 | 0.6 ± 0.3 | 0.9 ± 0.2 | 0.9 ± 0.3 | 1.0 ± 0.1 | 0.8 ± 0.2 |
20:5 n-3 | 16.1 ± 0.2 | 16.5 ± 0.4 | 18.7 ± 4.1 | 11.6 ± 0.2 | 9.8 ± 0.6 | 9.4 ± 1.0 |
22:5 n-3 | 0.8 ± 0.3 | 0.9 ± 0.2 | 0.6 ± 0.0 | 1.2 ± 0.1 | 1.1 ± 0.1 | 1.5 ± 0.3 |
22:6 n-3 | 9.0 ± 0.3 | 9.6 ± 1.4 | 8.6 ± 1.5 | 5.3 ± 0.1 | 4.3 ± 0.3 | 4.2 ± 0.4 |
Sum of PUFA | 32.3 ± 0.5 | 36.1 ± 1.4 | 37.9 ± 6.9 | 23.4 ± 0.5 | 21.3 ± 1.1 | 21.0 ± 1.6 |
Others | 2.0 ± 0.4 | 3.2 ± 0.3 | 3.7 ± 0.6 | 1.6 ± 0.1 | 1.5 ± 0.1 | 1.5 ± 0.1 |
n-3 | 29.9 ± 0.8 | 33.8 ± 1.5 | 35.5 ± 7.1 | 20.1 ± 0.3 | 18.1 ± 1.1 | 18.3 ± 1.2 |
n-6 | 2.4 ± 0.4 | 2.3 ± 0.2 | 2.3 ± 0.1 | 3.3 ± 0.3 | 3.3 ± 0.1 | 2.7 ± 0.3 |
TFA (pg cell−1) | 2.2 ± 0.1 | 2.0 ± 0.2 | 2.8 ± 0.3 | 2.5 ± 0.1 | 3.8 ± 0.5 | 6.9 ± 1.0 |
TFA (mg L−1) | 13.0 ± 1.7 | 17.2 ± 2.5 | 14.9 ± 1.9 | 57.7 ± 1.9 | 85.8 ± 4.2 | 71.9 ± 10.7 |
2.3. Increased TAG Accumulation and Oil-Body Formation Is to a Large Extent Attributable to Increased Carbon Availability
2.4. Increasing Inorganic Carbon Supply Enhances Accumulation of TAG Containing n-3 LC-PUFA
Bicarbonate (mM) | ||||||
---|---|---|---|---|---|---|
Before N-Limitation | After N-Limitation | |||||
Fatty Acids (% TFA) | 2 | 9 | 18 | 2 | 9 | 18 |
Saturated fatty acids | ||||||
14:0 | 11.2 ± 0.8 | 9.6 ± 0.9 | 9.6 ± 0.9 | 10.7 ± 0.6 | 9.7 ± 0.2 | 10.3 ± 0.4 |
16:0 | 22.6 ± 2.1 | 22.4 ± 1.3 | 22.4 ± 1.3 | 30.7 ± 0.8 | 27.4 ± 0.3 | 29.5 ± 0.5 |
18:0 | 5.6 ± 1.4 | 5.5 ± 1.7 | 5.5 ± 1.7 | 0.8 ± 0.1 | 0.9 ± 1.2 | 0.9 ± 0.2 |
Sum of SFA | 39.5 ± 0.1 | 37.5 ± 3.9 | 37.5 ± 3.9 | 42.2 ± 1.4 | 38.1 ± 0.3 | 40.7 ± 1.4 |
Monounsaturated fatty acids | ||||||
16:1 n-7 | 22.0 ± 3.3 | 22.9 ± 10.3 | 15.9 ± 0.3 | 36.5 ± 0.2 | 33.9 ± 0.1 | 30.8 ± 0.5 |
18:1 n-7 | 3.0 ± 0.9 | 4.5 ± 2.2 | 3.7 ± 0.5 | 2.0 ± 0.0 | 1.7 ± 0.1 | 1.8 ± 0.3 |
18:1 n-9 | 5.6 ± 2.1 | 4.5 ± 1.1 | 5.4 ± 0.8 | 2.1 ± 0.1 | 3.3 ± 0.2 | 2.4 ± 0.0 |
Sum of MUFA | 30.7 ± 2.1 | 31.8 ± 7.1 | 24.9 ± 0.6 | 40.7 ± 0.2 | 38.9 ± 0.2 | 35.0 ± 0.2 |
Polyunsaturated fatty acids | ||||||
18:2 n-6 | 3.5 ± 0.3 | 3.4 ± 0.3 | 3.9 ± 0.3 | 0.4 ± 0.1 | 0.4 ± 0.0 | 0.3 ± 0.0 |
18:3 n-6 | 0.2 ± 0.0 | 0.4 ± 0.0 | 0.8 ± 0.4 | 0.4 ± 0.0 | 0.6 ± 0.1 | 0.4 ± 0.1 |
18:3 n-3 | 1.2 ± 0.2 | 1.7 ± 0.2 | 2.0 ± 1.2 | 2.0 ± 0.0 | 2.4 ± 0.1 | 3.0 ± 0.2 |
18:4 n-3 | 1.5 ± 0.4 | 2.1 ± 0.3 | 1.7 ± 0.0 | 1.0 ± 0.1 | 1.5 ± 0.1 | 1.5 ± 0.1 |
20:4 n-6 | 1.3 ± 0.3 | 0.9 ± 0.2 | 1.6 ± 0.3 | 1.0 ± 0.1 | 1.1 ± 0.1 | 1.3 ± 0.1 |
20:5 n-3 | 7.6 ± 0.6 | 10.1 ± 1.4 | 10.8 ± 0.4 | 5.9 ± 0.5 | 7.9 ± 0.1 | 8.2 ± 0.5 |
22:5 n-3 | 0.4 ± 0.0 | 0.6 ± 0.0 | 0.7 ± 0.1 | 0.7 ± 0.1 | 1.1 ± 0.2 | 1.5 ± 0.1 |
22:6 n-3 | 3.5 ± 0.8 | 2.9 ± 0.1 | 1.4 ± 0.4 | 2.5 ± 0.5 | 4.2 ± 0.4 | 4.2 ± 0.3 |
Sum of PUFA | 19.3 ± 1.4 | 22.1 ± 2.2 | 23.0 ± 1.6 | 13.9 ± 1.3 | 19.2 ± 0.6 | 20.6 ± 1.2 |
Others | 10.6 ± 3.5 | 8.6 ± 1.0 | 16.9 ± 3.8 | 3.3 ± 0.3 | 3.8 ± 0.1 | 3.8 ± 0.1 |
n-3 | 14.3 ± 1.9 | 17.5 ± 1.8 | 16.7 ± 1.8 | 10.4 ± 1.2 | 15.3 ± 0.8 | 15.9 ± 0.9 |
n-6 | 5.0 ± 0.6 | 4.6 ± 0.4 | 6.3 ± 0.2 | 3.4 ± 0.1 | 3.9 ± 0.2 | 4.6 ± 0.3 |
TAG (pg cell−1) | 0.9 ± 0.1 | 0.7 ± 0.2 | 0.8 ± 0.2 | 1.3 ± 0.1 | 2.4 ± 0.1 | 4.8 ± 0.1 |
TAG (% TFA) | 42.9 ± 1.1 | 33.6 ± 6.1 | 28.5 ± 7.6 | 54.0 ± 1.6 | 68.3 ± 1.9 | 74.4 ± 6.4 |
3. Experimental Section
3.1. Strain and Culture Conditions
3.2. Growth Parameters, Medium pH, Biomass Harvesting and Storage
3.3. Nitrate Determination
3.4. Nile Red Staining and Microscopy
3.5. Lipid Extraction and Analysis
3.6. Fatty Acid Analysis
4. Conclusions
Abbreviations
DHA | docosahexaenoic acid |
EPA | eicosapentaenoic acid |
LC-PUFA | long-chain polyunsaturated fatty acids |
PUFA | polyunsaturated fatty acids |
TAG | triacylglycerols |
Acknowledgments
Conflicts of Interest
References
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Guihéneuf, F.; Stengel, D.B. LC-PUFA-Enriched Oil Production by Microalgae: Accumulation of Lipid and Triacylglycerols Containing n-3 LC-PUFA Is Triggered by Nitrogen Limitation and Inorganic Carbon Availability in the Marine Haptophyte Pavlova lutheri. Mar. Drugs 2013, 11, 4246-4266. https://doi.org/10.3390/md11114246
Guihéneuf F, Stengel DB. LC-PUFA-Enriched Oil Production by Microalgae: Accumulation of Lipid and Triacylglycerols Containing n-3 LC-PUFA Is Triggered by Nitrogen Limitation and Inorganic Carbon Availability in the Marine Haptophyte Pavlova lutheri. Marine Drugs. 2013; 11(11):4246-4266. https://doi.org/10.3390/md11114246
Chicago/Turabian StyleGuihéneuf, Freddy, and Dagmar B. Stengel. 2013. "LC-PUFA-Enriched Oil Production by Microalgae: Accumulation of Lipid and Triacylglycerols Containing n-3 LC-PUFA Is Triggered by Nitrogen Limitation and Inorganic Carbon Availability in the Marine Haptophyte Pavlova lutheri" Marine Drugs 11, no. 11: 4246-4266. https://doi.org/10.3390/md11114246