Effect of nitrogen limitation on growth, total lipid accumulation and protein amount in Scenedesmus acutus as biofuel reactor candidate

Abstract

Objective: In this study effects of nitrogen limitation on growth, total lipid and protein amount in Scenedesmus acutus.

Material and Methods: The microalgal strain of Scenedesmus acutus used in this study was isolated from the Keban Reservoir in Eastern Anatolia, Turkey was grown in Jaworsky’s medium. Algal cells were calculated by measuring the optical density at 680 nm using a visible density spectrophotometer. The total lipid content was determined using the Bligh and Dyer method. The total protein content was determined by the Lowry method.

Results: The results show that there is an inverse relationship between cellular growth, lipid amount and nitrogen concentration. S. acutus survived all the nitrogen concentrations tested and increases were observed in the amount of its cellular lipid. It was determined that there was enough nitrogen in the nitrogen-limited media to support protein synthesis and cell growth of S. acutus and that the amount of lipid in the 50% nitrogen-limited media was 19.48% higher than that in the control group.

Conclusion: S. acutus survived all the nitrogen concentrations tested and increases were observed in the amount of its cellular lipid. Significant increase in the amount of lipid in Scenedesmus acutus subjected to nitrogen stress suggests the idea that the microalga in question can be one of the potential organisms that can be used to obtain biofuel.

Keywords

• Scenedesmus acutus,nitrogen limitation,protein,lipid,biofuel

References

1. Ladanai S, Vinterback J. Global Potential of Sustainable Biomass for Energy. Uppsala: Swedish University of Agricultural Sciences. 2009; p32. doi:10.3390/en6020766.
2. 2. Chisti Y. Biodisel form microalgae. Biotechnol Adv. 2007; 25:294-306.
3. 3. Cardozo KHM, Guaratini T, Barros MP, Falcao VR, Tonon AP, Lopes NP, Campos S, Torres MA, Souza AO, Colepicolo P, Pinto E. Metabolites from algae with economical impact. Comp. Biochem. Phys. 2007; C.146:60-78.
4. 4. Huang G, Chen F, Wei D, Zhang X, Cgen G. Biodisel production by microalgal biotechnology. Appl. Energ. 2010; 87:38-46.
5. 5. Hu Q, Sommerfeld M, Jarvis E, Ghirardi M, Posewitz M, Seibert M, Darzins A. Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. Plant J. 2008; 54:621-639.
6. 6. Meng X, Yang J, Xu X, Zhang L, Nie Q, Xian M. Biodisel production from oleaginous microorganisms. Renew. Energ. 2009; 34:1-5.
7. 7. Chiu SY, Kao CY, Chen CH, Kuan TC, Ong SC, Lin CS. Reduction of CO2 by a high-density culture of Chlorella sp. In a semicontinuous photobioreactor. Bioresource Technol. 2008; 99:3389-3396.
8. 8. Li Y, Horsman M, Wang B, Wu N, Lan CQ. Effects of nitrogen sources on cell growth and lipid accumulation of green alga Neochlorisoleo abundans. Appl. Microbiol. Biot. 2008; 81:629-636.

9. 9. Yadavalli R, Ramogapol RS, Rao CS. Lipid Accumilation Studies in Chlorella pyreniodosa Using Customized Photobioreactor-Effect of Nitrogen Source, Light Intensity and Mode of Operatio. Int. J. Eng Research and Appl. 2012; 2(3):2446-2453.
10. 10. Wu H, Miau X. Biodiesel quality and biochemical changes of microalgae Chlorella pyrenodiosa and Scenedesmus obliquus in response to nitrate levels. Bioresource Technol. 2014; 170:421-427.
11. 11. Plumley FG, Schmidt GW. Nitrogen-dependent regulation of photosynthetic gene expression. Proc. Natl. Acad. Sci. USA. 1989; 86:2678-82.
12. 12. Li Y, Han D, Sommerfeld M, Hu Q. Photosynthetic carbon partitioning and lipid production in the oleaginous microalga Pseudochlorococcum sp. (Chlorophyceae) under nitrogen-limited conditions. Bioresource Technol. 2012; 102:123-129.
13. 13. Simionata D, Block MA, Rocca NL, Jouhet J, Marechal E, Finazzi G. The Response of Nannochloropsis gaditana to Nitrogen Starvation Includes De Novo Biosynthesis of Triacylgylcerols, a Decrease of Chloroplast Galactolipids, and Reorganization of the Photosynthetic Apparatus. E.C Journal ASMorg. 2013; 12(5):665-676.
14. 14. Iliman AM, Scagg AH, Shales SW. Increase in Chlorella strains calorific values when grown in low nitrogen medium. Enzyme Microb Tech. 2000; 27:631-635.
15. 15. Agirman N, Cetin AK. Effects of Nitrogen Starvations on Cell Growth, Protein and Lipid Amount of Chlorella vulgaris. Fresen Environ Bull. 2015; 24 (11):3643-3648.
16. 16. Converti A, Casazza AA, Ortiz EY, Perego P, Borghi MD. Effect of temperature and nitrogen concentration on the growth and lipid content of Nannochloropsis oculata and Chlorella vulgaris. Chem. Eng. Prog. 2009; 48:1146-51.
17. 17. Bligh EG, Dyer WJ. A rapid method for total lipid extraction and purification. Can. Biochem. Physiol. 1959; 37:911-917.
18. 18. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol regent. J. Biol. Chem. 1951; 193:265-75.
19. 19. Negi S, Barry AN, Friedland N, Sudasinghe N, Subramanian S, Pieris S, Holguin FO, Dungan B, Schaub T, Sayre R. Impact of Nitrogen limitation on biomass, photosynthesis, and lipid accumulation in Chlorella sorokiniana. J Appl. Phycol. 2016; 28:803-812.
20. 20. Ikaran Z, Alvarez SS, Castanon US. The effect of nitrogen limitation on the physiology and metabolism of Chlorella vulgaris var L3. Algal Res. 2015; 10:134-144.
21. 21. Saha SK, Mchugan E, Hayes J, Moane S, Walsh D, Murray P. Effect of various stress-regulatory factors on biomass and lipid production in microalga Haematococcus pluvialis. Bioresource Technol. 2013. 128:118-124.
22. 22. Griffiths MJ, Harrison STL. Lipid productivity as a key characteristic for choosing algal species for biodiesel production. J. Appl. Phycol. 2009; 21:493-507.
23. 23. Pal D, Khozin-Goldberg I, Cohen Z, Boussiba S. The effect of light, salinity, and nitrogen availability on lipid production by Nannochloropsis sp. Appl. Microbiol. Biot. 2011; 90:1429-1441.
24. 24. Huang X, Huang Z, Wen W, Yan J. Effects of nitrogen supplementation of the culture medium on the growth, total lipid content and fatty acid profiles of three microalgae Tetraselmis subcordiformis, Nannochloropsis oculata and Pavlova viridis. J. Appl. Phycol. 2013; 25:129-37.
25. 25. Yeesang C, Cheirsilp B. Effect of nitrogen, salt, and iron content in the growth medium and light intensity on lipid production by microalgae isolated from freshwater sources in Thailand. Bioresource Technol. 2011; 102:3034-3040.
26. 26. Widjaja A, Chien CC, Ju YH. Study of increasing lipid production from fresh water microalgae Chlorella vulgaris. J. Taiwan Ins. Chem Eng. 2009; 40:13-20.
27. 27. Dean AP, Sigee DC, Estrada B, Pittman JK. Using FTIR spectroscopy for rapid determination of lipid accumulation in response to nitrogen limitation in freshwater microalgae. Bioresource Technol. 2010; 101:4499–4507.
28. 28. Bottam PA, Ratledge C. A Biochemical Explanation for Lipid Accumulation in Candida 107 and Other Oleaginous Micro-organisms. J. Gen. Microbiol. 1979; 114:361-371.
29. 29. Chen F, Johns MR. Effect of C/N ratio and aeration on the fatty acid composition of heterotrophic Chlorella sorokiniana. J. Appl. Phycol. 1991; 3:203-209.
30. 30. Ratledge C, Wynn JP. The biochemistry and biology of lipid accumulation in oleaginous Microorganisms. Adv. Appl. Microbiol. 2002; 51:1-51.
31. 31. De Swaaf ME, Sijtsma L, Pronk JT. High-Cell-Density Fed-Batch Cultivation of the Docosahexaenoic Acid Producting Marine Alga Crypthecodinium cohnii. Biotechnol. Bioeng. 2003 ; 81:666-672.
32. 32. Garcia-Ferris C, De Los Rios A, Moreno J. Correlated biochemical and ultrastructural changes in nitrogen starved Euglena gracilis. J. Phycol. 1996 ; 32:953-963.
33. 33. Sheehan J, Dunahay T, Benemann J, Roessler P. A look back at the U.S. Department of Energy’s aquatic species program-biodiesel from algae. Golden: National Renewable Energy Laboratory. 1998; NREL/TP-580-24190.
34. 34. Ganuza E, Anderson AJ, Ratledge C. High-cell-density cultivation of Schizochytrium sp. in an ammonium/pH-auxostat fed-batch system. Biotechnol. Lett. 2008; 30:1559-1564.
35. 35. Breuer G, Lamers PP, Martens DE, Draaisma RB, Wijffels RH. The impact of nitrogen starvation on the dynamics of triacylglycerol accumulation in nine microalgae strains. Bioresource Technol. 2012; 124:217-226.
36. 36. Li YX, Zhao FJ, Yu DD. Effect of Nitrogen Limitation on Cell Growth, Lipid Accumulation and Gene Expression in Chlorella sorokiniana. Braz Arch Biol Technol. 2015; 58(3): 462-7.
37. 37. Guerrini F, Cangini M, Boni L, Trost P, Pistocchi R. Metabolic responses of the diatom Achnanthes brevipes (Bacillariophyceae) to nutrient limitation. J. Phycol. 2000 ; 36:882-890.
38. 38. Gladue RM, Maxey JE. Microalgal feeds for aquaculture. J Appl. Phycol. 2009; 6:131-141.