Effects of nitrogen to phosphorus molar ratios on the growth and fatty acid accumulation of a freshwater microalga Chlorococcum humicola
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Abstract
Microalgae cultivation for being a biological feedstock of high-value products has long depended on culture conditions and various important factors. The objective of this research was to investigate the effects of nitrogen to phosphorus molar ratios on the growth and fatty acid accumulation of a freshwater microalga Chlorococcum humicola (TISTR 8551). The microalgae having an initial density of 2.00x106 cell mL-1 was cultivated in a bubble column photobioreactor with a working volume of 10 L under batch condition for 14 days. The growth broth employed was modified BG-11 medium with the initial molar ratios of nitrogen to phosphorus (N:P) of 50:1 (GM-100%N), 5:1 (GM-10%N), and 2.5:1 (GM-5%N). It was found that the growth and fatty acid of the microalgae had different responses to a variation of the initial nitrogen concentration of the growth medium. The microalgae cultivated in the GM-100%N provided maximum biomass productivity of 0.77 ± 0.14 g L-1 d-1 and a specific growth rate () of 0.139 d-1. The figure of obtained was higher than those cultivated in the GM-10%N and GM-5%N with 69.51 and 98.57%, respectively, because the GM-100%N had enough concentrations of nitrogen for the microalgae to synthesize protein and chlorophyll. Then, these compounds were used by the microalgae for photosynthesis and cell division resulted in lower activities of fatty acid synthesis and accumulation than those cultivated in the growth medium having relatively lower initial N:P molar ratios. On the other hand, the microalgae cultivated in the GM-5%N gave the highest cumulative fatty acid content of 30.6 ± 1.2 %DCW. This was because the GM-5%N provided a higher degree of nitrogen starvation. Therefore, the microalgae reduced protein and chlorophyll synthesis activities and turned to increase fatty acid synthesis and accumulation in cells to prevent cell damage and degradation from light exposure during the cultivation period.
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References
Benavente-Valdés JR, Aguilar C, Contreras-Esquivel JC, Mendez-Zavala A, Montanez J. Strategies to enhance the production of photosynthetic pigments and lipids in chlorophycae species. Biotechnology Reports. 2016; 10: 117-125.
Chisti Y.Biodiesel from microalgae.Biotechnology Advances. 2007; 25: 294-306.
Goiris K, Colen WV, Wilches I, León-Tamariz F, Cooman LD, Muylaert K. Impact of nutrient stress on antioxidant production in three species of microalgae.Algal Research. 2015; 7: 51-57.
Khozin-Goldberg I, Cohen Z. The effect of phosphate starvation on the lipid and fatty acid and composition
of the fresh water eustigmatophyte Monodus subterraneus. Phytochemistry. 2006; 67: 696-701.
Xin L, Hong-Ying H, Ke G, Ying-Xue S. Effect of different nitrogen and phosphorus concentrations on the growth, nutrient uptake and lipid accumulation of a freshwater microalgae Scenedesmus sp. Bioresource Technology. 2010; 191: 5494-5500.
Vymazal J. Algae and Element Cycling in Wetland. Boca Raton: Lewis Publishers; 1995.
Perez-Gacia O, Escalanie FME, Bashan LED, Bashan Y. Heterotrophic cultures of microalgae: Metabolism and potential products. Water Resources. 2011; 45: 11-36.
Morales-Sánchez D, Kyndt J, Ogden K, Martinez A. Toward an understanding of lipid and starch accumulation in microalgae: A proteomic study of Neochloris oleoabundans cultivated under N-limited heterotrophic conditions. Algal Research. 2016; 20:
-34.
Bona F, Capuzzo A, Francino M, Maffel ME.
Semi-continuous nitrogen limitation as convenient operation strategy to maximize fatty acid production in Neochloris oleoabundans. Algal Research. 2014; 5: 1-6.
Lari Z, Moradi-Kheibari N, Ahmadzadeh H, Abrishamchi P, Moheimani NR, Murry MA.Bioprocess engineering of microalgae to optimize lipid production through nutrient management. Journal of Applied Phycology. 2016; 28: 3235-3250.
Najafabadi HA, Malekzadeh M, Jalilian F, Vossoughi M, Pazuki G. Effect of various carbon sources on biomass and lipid production of Chlorella vulgaris during nutrient sufficient and nitrogen starvation conditions. Bioresource Technology. 2015; 180: 311-317.
Singh P, Guldhe A, Kumari S, Rawat I, Bux F. Combined metals and EDTA control: An integrated and scalable lipid enhancement strategy to alleviate biomass constraints in microalgae under nitrogen limited conditions. Energy Conversion and Management. 2016; 114: 100-109.
Phankosol S, Chum-in T, Krisnangkura K. Free energy additivity method for prediction dynamics viscosity of vegetable oils at various temperatures to its chemical compositions. UBU Engineering Journal. 2017; 10(2):
-51.
Pokethitiyook P, Yuan JP, Meetam M, Sritong K, Pugkaew W, Damrongphol P. Culture of microalgal strains isolated from natural habitats in Thailand in various enriched media. Applied Energy. 2012; 89(1): 296-302.
Kirtania K, Bhattacharya S. Application of the distributed activation energy model to the kinetic study of pyrolysis of the fresh water algae Chlorococcum humicola. Bioresource Technology. 2012; 107: 476-481.
Griffiths MJ, Harrison STL. Lipid productivity as a key characteristic for choosing algal species for biodiesel production. Journal of Applied Phycology. 2009; 21(5): 493-507.
Boussiba A, Vonshak A. Astaxanthin accumulation in
the green alga Haematococcus pluviaris. Plant Cell Physiology. 1991; 32(7): 1077-1082.
Wannasutthiwat S. Growth and Enhancement of Carotenoids Production in Microalga Chlorococcum humicola in Continuous Condition. MEng Thesis. Bangkok: Graduate school, Chulalongkorn University; 2014.
Kunyawut C, Paopo I, Krommuang A. Development of a bubble photobioreactor for microalgal culture. Burapha Science Journal. 2019; 24(2): 471-488.
Doran, PM. Bioprocess Engineering Principles. 2nd ed. London: Elsevier; 2013.
Thiangpakdee N, Kunyawut C, Paopo I. An increase
in domestic wastewater treatment capacity of Chlorococcum humicola. Burapha Science Journal. 2021; 26(2): 1279-1292.
Fan LS. Gas Liquid-Solid Fluidization Engineering.
London: Butterworth-Heinemann; 1989.
Krichnavaruk S, Powtongsook S, Pavasant P. Enhanced production capacity of Chaetoceros calcitrans in airlift photobioreactors. Bioresource Technology. 2007; 98: 2123-2130.
APHA. Standard Methods for the Examination of Water and Wastewater. Washington, DC: American Water Works Association and Water Pollution Control Federation; 1999.
Strickland JDH, Parsons TR. A Practical Handbook of Seawater Analysis. Toronto. Fisheries Research Board of Canada: Alger Press; 1972.
Bright EG, Dyer WJ. A rapid method of total liquid extraction and purification.Canadian Journal of Biochemistry and Physiology. 1958; 37(8): 911-917.
Vaičiulyte S, Padovani G, Kostkevičiene J, Carlozzi, P. Batch growth of Chlorella Vulgaris CCALA 869 versus semi-continuous regimen for enhancing oil-rich biomass productivity. Energies. 2014; 7: 3840-3857.
Krichnavaruk S, Loataweesup W, Powtongsook S, Pavasant P. Optimal growth conditions and cultivation of Chaetoceros calcitrans in airlift photobioreactor. Chemical Engineering Journal. 2005; 105: 91-98.
ÖrdÖg V, Stirk WA, Balint P, Staden JV, Lovase, C.
Change in lipid, protein and pigment concentrations in
nitrogen-stressed Chlorella minutissima.Journal of Applied Phycology. 2012; 24: 907-914.
Yang B, Jian JF, Kuo CM, Zhang DH, Lin CS. Biomass
and lipid production of Chlorella sp. using municipal wastewater under semi-continuous cultivation. International Proceedings of Chemical, Biological and Environmental Engineering. 2017; 101: 485-493.
Grobbelaar JU. Algal nutrient: Mineral nutrient. In: Amos Richmond, editor.Handbook of Microalgal Culture: Biotechnology and Applied Phycology.London:
Wiley-Blackwell; 2004.
Redfield AC. The biological content of chemical factors in the environment.American Scientist.1958; 46:
-221.
Arbib Z, Ruiz J, Alvarez-Dias P, Garrido-Perez C, Barragan J, Perales JA. Photobiotreatment: influence of nitrogen and phosphorus Ratio in wastewater on growth kinetics of Scenedesmus obliquus.International journal of phytoremediation. 2013; 15(8): 774-788.
Feng P, Deng Z, Fan L, Hu Z. Lipid accumulation and growth characteristics of Chlorella zofingiensis under different nitrate and phosphate concentrations. Journal of Bioscience and Bioengineering. 2012; 114(4): 405-410.
Monfet E, Unc A. Defining wastewaters used for cultivation of algae. Algal Research. 2017; 24: 520-528.
Krommuang A, Kunyawut C, Paopo I.Domestic wastewater treatment with microalgae Chlorococcum humicola in an airlift photobioreactor.Journal of Industrial Technology Ubon Ratchathani Rajabhat University. 2021; 11(1): 55-57.
Gu W, Li H, Zhao P, Yu R, Pan G, Gao S, Xie X, Huang A, He L, Wang G.Quantitative proteomic analysis of thylakoid from two microalgae Haematococcus pluvialis and Dunaliella salina reveals two different high
light-responsive strategies. Scientific Reports. 2014; 6661: 1-12.
Khoo CG, Lam MK, Lee KT. Pilot-scale semi-continuous cultivation of microalgae Chlorella Vulgaris in bubble column photobioreactor (BC-PBR): Hydrodynamics and gas–liquid mass transfer study. Algal Research, 2016; 15: 65-76.
Adams C, Godfrey V, Wahlen B, Seefeldt L, Bugbee B. Understanding precision nitrogen stress to optimize the growth and lipid content tradeoff in oleaginous green microalgae. Bioresource Technology. 2013; 131: 188-194.
Liu T, Li Y, Liu F, Wang C. The enhanced lipid accumulation in oleaginous microalga by the
potential continuous nitrogen-limitation (CNL) strategy. Bioresource Technology. 2016; 203: 150-159.
Pancha I, Chosh A, Mishra S. Salinity induced oxidative stress alters the physiological responses and
improves the biofuel potential of green
microalgae Acutodesmus dimorphus.Bioresource Technology. 2017; 244, 1376-1383.
Mousavi S, Najafpour GD, Mohammadi M. CO2 bio-fixation and biofuel production in an airlift photobioreactor by an isolated strain of microalgae Coelastrum sp. under high CO2 concentration.Environmental Science and Pollution Research. 2018; 25: 30139-30150.
Wang HT, Meng YY, Cao XP, Ai JN, Zhou JN, Xue S, Wang WL. Coordinated response of photosynthesis, carbon assimilation, and triacylglycerol accumulation to nitrogen starvation in the marine microalgae Isochrysis zhangjiangensis (haptophyta). Bioresource Technology. 2015; 177: 282-288.
Cabanelas ITD, Kleinegris DMM, Wijffels RH, Barbosa MJ. Repeated nitrogen starvation doesn’t affect lipid productivity of Chlorococcum littorale. Bioresource Technology. 2016; 219: 576-582.
Shen XF, Liu JJ, Chauhan AS, Hu H, Ma LL, Lam PKS,
Zeng RJ.Combining nitrogen starvation with sufficient phosphorus supply for enhanced biodiesel productivity of Chlorella vulgaris fed on acetate. Algal Research. 2016; 17: 261-267.