Effect of gradual nitrogen removal on growth, biochemical composition and fatty acid profile of microalga Haematococcus pluvialis

Document Type : Research Paper

Authors

1 M.Sc. in Plant Physiology, Department of Biology, College of Science, Shiraz University, Shiraz, Iran

2 Assistant Professor in Department of Biology, College of Science, Shiraz University, Shiraz, Iran

10.22124/japb.2022.22780.1476

Abstract

Different physicochemical factors influence the growth of Haematococcus pluvialis and should be assessed in order to optimal production of biomass and its chemical composition. There is few information on the effect of gradual nitrogen limitation on the microalgae.  Hence, in the present study the effects of gradual nitrogen removal from culture medium, in two stages, on growth, chlorophyll and astaxanthin content, biochemical compounds and fatty acid profile in H. pluvialis was evaluated. During 55 days culture, biomass production and chlorophyll content in the treatment group showed a significant decrease (P<0.05) compared to the control, whilst no significant change was observed in the astaxanthin content. The highest content of astaxanthin was observed before the replacement of culture medium in both experimental groups. At the end of the experiment, the content of lipid, protein, and carbohydrate in the nitrogen-free culture showed a significant decrease. Fatty acid profile showed a reduction in unsaturated fatty acid with nitrogen limitation. Overall, the results showed that the stepwise nitrogen depletion caused a change in biochemical composition of microalgae. In the same condition, as the algal cells were gradually subjected to severe stress, consequently, the production of astaxanthin was not affected.

Keywords

Main Subjects

References

Bradford M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2): 248–254.
Breuer G., Evers W.A., De Vree J.H., Kleinegris D.M., Martens D.E., Wijffels R.H. and Lamers P.P. 2013. Analysis of fatty acid content and composition in microalgae. Journal of Visualized Experiments, 80: 1–9 (e50628).
Cakmak T., Angun P., Demiray Y.E., Ozkan A.D., Elibol Z. and Tekinay T. 2012. Differential effects of nitrogen and sulfur deprivation on growth and biodiesel feedstock production of Chlamydomonas reinhardtii. Biotechnology and Bioengineering, 109: 1947–1957.
Chai S., Shi J., Huang T., Guo Y., Wei J., Guo M., Li L., Dou S., Liu L. and Liu G. 2018. Characterization of Chlorella sorokiniana growth properties in monosaccharide-supplemented batch culture. PLoS One, 13: 1–19 (e0199873).
Chia M.A., Lombardi A.T., Melao M.D.G. and Parrish C.C. 2013. Effects of cadmium and nitrogen on lipid composition of Chlorella vulgaris (Trebouxiophyceae, Chlorophyta). European Journal of Phycology, 48(1): 1–11.
Cobos M., Paredes J.D., Maddox J.D., Vargas-Arana G., Flores L., Aguilar C.P., Marapara J.L. and Castro J.C. 2017. Isolation and characterization of native microalgae from the Peruvian Amazon with potential for biodiesel production. Energies, 10: 1–16.
Courchesne N.M.D., Parisien A., Wang B. and Lan C.Q. 2009. Enhancement of lipid production using biochemical, genetic and transcription factor engineering approaches. Journal of Biotechnology, 141: 31–41.
Da Silva A.F., Lourenco S.O. and Chaloub R. 2009. Effects of nitrogen starvation on the photosynthetic physiology of a tropical marine microalgae Rhodomonas sp. (Cryptophyceae). Aquatic Botany, 91(4): 291–297.
Dubois M., Gilles K., Hamilton J., Rebers P. and Smith F. 1956. Phenol sulphuric acid method for total carbohydrate. Analytical Chemistry, 26: 350–356.
Geider R.J., Roch J.L., Green R.M. and Olaizola M. 1993. Response of the photosynthetic apparatus of Phaeodactylum tricornutum (Bacillariophyceae) to nitrate, phosphate, or iron saturation. Journal of Phycology, 29: 755–766.
Hermansyah H., Sakinah R.A., Julinar J., Hanafiah Z. and Zulkifli H. 2018. Bioethanol production from microalgae Oscillatoria sp. cultured in blue green 11 and bold basal media. E3S Web of Conferences, 68: 1–8 (03018).
Jannel S., Caro Y., Bermudes M. and Petit T. 2020. Novel insights into the biotechnological production of Haematococcus pluvialis-derived astaxanthin: Advances and key challenges to allow its industrial use as novel food ingredient. Journal of Marine Science and Engineering, 8(10): 1–48 (789).
Kobayashi N., Noel E.A., Barnes A., Watson A., Rosenberg J.N., Erickson G. and Oyler G.A. 2013. Characterization of three Chlorella sorokiniana strains in anaerobic digested effluent from cattle manure. Bioresour Technology, 150: 377–386.
Li F., Cai M., Lin M., Huang X., Wang J., Ke H. and Yang S. 2020. Enhanced biomass and astaxanthin production of Haematococcus pluvialis by a cell transformation strategy with optimized initial biomass density. Marine Drugs, 18(7): 1–18 (341).
Li F., Cai M., Wu Y., Lian Q., Qian Z., Luo J., Zhang Y., Zhang N., Li C. and Huang X. 2022. Effects of nitrogen and light intensity on the astaxanthin accumulation in motile cells of Haematococcus pluvialis. Frontiers in Marine Science. 9: 1–7 (909237).
Lichtenthaler H.K. and Buschmann C. 2001. Chlorophylls and carotenoids: Measurement and characterization by UV‐VIS spectroscopy. Current Protocols in Food Analytical Chemistry, 1(1): 1–8 (F4.3).
Liu T., Chen Z., Xiao Y., Yuan M., Zhou C., Liu G., Fang J. and Yang B. 2022. Biochemical and morphological changes triggered by nitrogen stress in the oleaginous microalga Chlorella vulgaris. Microorganisms, 10(3): 1–16 (566).
Liyanaarachchi V.C., Nishshanka G.K.S.H., Premaratne R.G.M. M., Ariyadasa T.U., Nimarshana P.H.V. and Malik A. 2020. Astaxanthin accumulation in the green microalga Haematococcus pluvialis: Effect of initial phosphate concentration and stepwise/continuous light stress. Biotechnology Reports, 28: 1–11 (e00538).
Msanne J., Xu D., Konda A.R., Casas-Mollano J.A., Awada T., Cahoon E.B. and Cerutti H. 2012. Metabolic and gene expression changes triggered by nitrogen deprivation in the photoautotrophically grown microalgae Chlamydomonas reinhardtii and Coccomyxa sp.           C-169. Phytochemistry, 75: 50–59.
Ordog V., Stirk W.A., Balint P., Van Staden J. and Lovasz C. 2012. Changes in lipid, protein and pigment concentrations in nitrogen-stressed Chlorella minutissima cultures. Journal of Applied Phycology, 24: 907–914.
Oslan S.N. H., Shoparwe N.F., Yusoff A.H., Rahim A.A., Chang C.S., Tan J.S. and Sulaiman A.Z. 2021. A Review on Haematococcus pluvialis bioprocess optimization of green and red stage culture conditions for the production of natural astaxanthin. Biomolecules, 11(2): 1–15 (256).
Panis G. and Carreon J.R. 2016. Commercial astaxanthin production derived by green alga Haematococcus pluvialis: A microalgae process model and a techno-economic assessment all through production line. Algal Research, 18: 175–190.
Recht L., Zarka A. and Boussiba S. 2012. Patterns of carbohydrate and fatty acid changes under nitrogen starvation in the microalgae Haematococcus pluvialis and Nannochloropsis sp. Applied Microbiology and Biotechnology, 94(6): 1495–1503.
Saha S.K., McHugh E., Hayes J., Moane S., Walsh D. and Murray P. 2013. Effect of various stress-regulatory factors on biomass and lipid production in microalga Haematococcus pluvialis. Bioresource Technology, 128: 118–124.
Sathasivam R., Pongpadung P., Praiboon J., Chirapart A., Trakulnaleamsai S., Roytrakul S. and Juntawong N. 2018. Optimizing NaCl and KNO3 concentrations for high β-carotene production in photobioreactor by Dunaliella salina KU11 isolated from saline soil sample. Chiang Mai Journal of Science, 45: 106–115.
Shah M., Mahfuzur R., Liang Y., Cheng J.J. and Daroch M. 2016. Astaxanthin-producing green microalga Haematococcus pluvialis: From single cell to high value commercial products. Frontiers in Plant Science, 7: 1–28 (531).
Tarazona-Delgado R., Guarieiro M.D., Antunes P.W., Cassini S.T., Terreros H.M. and Fernandes V.O.  2021. Effect of nitrogen limitation on growth, biochemical composition, and cell ultrastructure of the microalga Picocystis salinarum. Journal of Applied Phycology, 33: 2083–2092.
Yaakob M.A., Mohamed R.M.S.R., Al-Gheethi A., Aswathnarayana Gokare R. and Ambati R.R. 2021. Influence of nitrogen and phosphorus on microalgal growth, biomass, lipid, and fatty acid production: An overview. Cells, 10: 393–412.
Young E.B. and Beardall J. 2003. Rapid ammonium- and nitrate-induced perturbations to Chl a fluorescence in nitrogen-stressed Dunaliella tertiolecta (Chlorophyta). Journal of Phycology, 39: 332–342.
Zhang Y., Wu H., Yuan C., Li C.T. and Li A. 2019. Growth, biochemical composition, and photosynthetic performance of Scenedesmus acuminatus during nitrogen starvation and resupply. Journal of Applied Phycology, 31: 2797–2809.
Zhao L.S., Li K., Wang Q.M., Song X.Y., Su H.N., Xie B.B., Zhang X.Y., Huang F., Chen X.L., Zhou B.C. and Zhang Y.Z. 2017. Nitrogen starvation impacts the photosynthetic performance of Porphyridium cruentum as revealed by chlorophyll a fluorescence. Scientific Reports, 7(1): 8542–8553.
Zhao Y., Yue C., Geng S., Ning D., Ma T. and Yu X. 2019. Role of media composition in biomass and astaxanthin production of Haematococcus pluvialis under two-stage cultivation. Bioprocess and Biosystems Engineering, 42: 593–602.
Zhu S., Huang W., Xu J., Wang Z., Xu J. and Yuan Z. 2014. Metabolic changes of starch and lipid triggered by nitrogen starvation in the microalga Chlorella zofingiensis. Bioresource Technology, 152: 292–298.
Zhu S., Wang Y., Shang C., Wang Z., Xu J. and Yuan Z. 2015. Characterization of lipid and fatty acids composition of Chlorella zofingiensis in response to nitrogen starvation. Journal of Bioscience and Bioengineering, 120(2): 205–209.
Aquatic Physiology and Biotechnology
Volume 11, Issue 2
September 2023
Pages 95-114

Files

Share

How to cite

Statistics