rapid induction of omega 3 fatty acid

1. Rapid Induction of Omega-3 Fatty Acids
(EPA) in Nannochloropsis sp. by UV-C
Radiation

2. Impact factor 4.24

3. Omega-3 Fatty Acid:
Dietary Fat
Saturated Fat Unsaturated Fat
MUFA PUFA
ω-3 FA ω-6 FA
ALA EPA DHA

4. Functions of Omega-3 FA:
 Boost brain development.
 Reduce risk of cardiovascular diseases.
 Reduce inflammation.
 Helps in blood pressure regulation.
 Required for the normal development and function
of retina.

5. Nannochloropsis sp.
 Systematic position:
Domain : Eukaryota
Super-phylum : Heterokonta
Class : Eustimatophyceae
Family : Eustigmaceae
Genus : Nannochloropsis
 Six known species.
 Small non-motile spheres
 Chlorophyll a is present and chlorophyll b&c absent.
 About 2-3 µm in diameter.
 Abiliy to accumulate high levels of PUFA.

6. Overview:

7. Techniques used:
Flowcytometry:
 To detect, identify, analyses and sort out different
population of cells.
Gas chromatography coupled mass spectrometry (GC-
MS):
 Gas chromatography mass spectrometry (GC-MS) is an
instrumental technique, comprising a gas chromatograph
(GC) coupled to a mass spectrometer (MS), by which
complex mixtures of chemicals may be separated,
identified and quantified.

8. Flow cytometry:

9. GC-MS (Gas chromatography coupled
mass spectrometry)

10. Materials and Methods:
 Sample collection.
 Culturing
 Culture subjected to UV-C stress.
 Lipid fluorescence analysis.
 GC-MS analysis
 Dry weight measurement.
 Analytical methods.

12. Nile red stained Nannochloropsis cells
subjected to different UV-C dosages

13. Kill curve showing the no. of cells survived
with increased UV intensities.

14. FACS analysis

15. Graphical representation of FACS
analysis
UV-C treatment (mj/cm²)

16. Lipid induction in Nannochloropsis sp. at
24 h after receiving different UV dosage

17. Fatty acid profiling in Nannochloropsis
following UV-C radiation shows lipid
induction and a shift towards LC-PUFAs
 To further quantify the ability of UV-C stress to
increase cellular fatty acid contents and to profile their
composition, GC-MS analysis was carried out.
 These confirmed the results obtained from flow
cytometry, showing a significant total fatty acid
increase of the UV-C treated culture as compared to
control culture.
 Moreover in cultures treated with UV-C stress, the
amount of USFA significantly increased and also the
proportion of USFA to the total fatty acids.

18. Cont.

19. Comparative increase in different SFAs
and USFAs after UV-C stress.

20. Dry weight proportion changes in
Nannochloropsis following nutrition stress
and different UV-C dosage

21. EPA proportion changes in Nannochloropsis
following nutrient starvation and different UV-C
dosage

22. Discussion:
 A low dosage of 100 & 250 mj/cm² was optimal to induce
total lipid accumulation espacially PUFA.
 A notable difference when comparing the flow-
cytometry and GC-MS was found.
 Linear decline in the total dry weight seen.
 Significant increase in the % of EPA compared to the
control with in increasing UV-C intensity.
 UV-C treatment is fast and get result in 24 h.
 Omega-3 fatty acid (EPA) significantly increased.
 Antioxidant role of PUFA.
 EPA also an important part of cell membrane of
microalgae and plays important role in cellular repair
and membrane development.

23. Conclusion:
 This study higlights the use of low exposure of UV-C
radiation at 100 and 250 mj/cm² to induce lipid
accumulation in Nannochloropsis particularly EPA
within 24h of the treatment.
 The optimized large-scale production of EPA in
Nannochloropsis may provide much needed
alternative, vegetarian source of Omega-3.
 Suggests that genetic manipulation can be done in
future aimed at the genetic improvement of the
current oleaginous strains.