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Influence of nutrient formulations on growth, lipid yield, carbon partitioning and biodiesel quality potential of Botryococcus sp. and Chlorella sp.

Environmental Science and Pollution Research volume 26, pages 7589–7600 (2019)Cite this article

Abstract

The study was conducted to analyse the influence of three nutrient formulations, namely BG-11 medium, BBM and TAP medium, on growth potential and lipid yield of two microalgal genera (Botryococcus sp. and Chlorella sp.) and to study the roles of N, P and other major nutrients. The study focussed on the general patterns of starch and lipid synthesis and storage and to further assess how photosynthetic carbon partitioning into starch and lipid is altered by conditions in growth media such as N and C presence as seen in BG11 medium which are known to induce neutral lipid production and the lack of it in BBM and TAP medium. BG-11 medium performed better as compared to BBM and TAP medium in terms of biomass productivity and lipid yield. The lipid yield was highest in Botryococcus sp. (63.03% dry wt.) and Chlorella sp. (50.27% dry wt.) at 30th day of incubation. Mean biomass productivity was highest for Botryococcus in BBM medium (6.14 mg/L/day) and for Chlorella in BG-11 medium (4.97 mg/L/day). Mean lipid productivity (50.78% and 39.36%) was highest in BG11 medium for both Botryococcus and Chlorella species, respectively. A sharp decline in sugar content was observed in the late stationary phase of growth from 30th day to 45th day. Fatty acid methyl ester (FAME) profile of the extracted lipids showed predominantly oleic acid, followed by palmitic acid and stearic acid in both the strains when grown in BG-11 medium. The other biodiesel quality parameters were in accordance with the international standards. A complex relationship was found between chemical composition and biodiesel properties. Proximity analysis indicated that the fuel properties of biodiesels are determined by a number of parameters and by the combination of different chemical compositions. The results provide an insight into organic carbon partitioning into lipid compounds and how the organism’s lipid metabolism changes due to N-deplete culturing in TAP medium and inorganic carbon source availability as seen in BG-11 and BBM medium.

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References

  • Adams C, Godfrey V, Wahlen B, Seefeldt L, Bugbee B (2013) Understanding precision nitrogen stress to optimize the growth and lipid content trade-off in oleaginous green microalgae. Bioresour Technol 131:188–194

    Article  CAS  Google Scholar 

  • Baig RN, Verma S, Nadagouda MN, Varma RS (2016) Room temperature synthesis of biodiesel using sulfonated graphitic carbon nitride. Sci Rep 6:39387

    Article  CAS  Google Scholar 

  • Behrens PW, Kyle DJ (1996) Microalgae as a source of fatty acids. J Food Lipids 3:259–272

    Article  CAS  Google Scholar 

  • Bhola V, Desikan R, Santosh SK, Subburamu K, Sanniyasi E, Bux F (2011) Effectsof parameters affecting biomass yield and thermal behaviour of Chlorella vulgaris. J Biosci Bioeng 111(3):377–382

    Article  CAS  Google Scholar 

  • Bligh EG, Dyer WJ (1959) Extraction of lipids in solutions by method of lipid extraction. Can J Biochem Physiol 37:911–917

    Article  CAS  Google Scholar 

  • Brányiková I, Maršálková B, Doucha J, Brányik T, Bišová K, Zachleder V, Vítová M (2011) Microalgae—novel highly efficient starch producers. Biotechnol Bioeng 108:766–776

    Article  CAS  Google Scholar 

  • Briat JF, Dubos C, Gaymard F (2015) Iron nutrition, biomass production and plant product quality. Trends Plant Sci 20:33–40

    Article  CAS  Google Scholar 

  • Calixto CD, da Silva Santana JK, Tibúrcio VP, da Costa Sassi CF, da Conceição MM, Sassi R (2018) Productivity and fuel quality parameters of lipids obtained from 12 species of microalgae from the northeastern region of Brazil. Renew Energy 115:1144–1152

    Article  CAS  Google Scholar 

  • Canaki M, Sanli H (2008) Biodiesel production from various feed stocks and their effects on the fuel properties. J Ind Microbiol Biotechnol 35:431–441

    Article  CAS  Google Scholar 

  • Chen CY, Yeh KL, Aisyah R, Lee DJ, Chang JS (2011) Cultivation, photobioreactor design and harvesting of microalgae for biodiesel production: a critical review. Bioresour Technol 102(1):71–81

    Article  CAS  Google Scholar 

  • Cheng P, Wang J, Liu T (2014) Effects of nitrogen source and nitrogen supply model on the growth and hydrocarbon accumulation of immobilized biofilm cultivation of B. braunii. Bioresour Technol 166:527–533

    Article  CAS  Google Scholar 

  • Chisti Y (2007) Biodiesel from microalgae. Biotechnol Adv 25:294–306

    Article  CAS  Google Scholar 

  • Converti A, Casazza AA, Ortiz EY, Perego P, Del Borghi M (2009) Effect of temperature and nitrogen concentration on the growth and lipid content of Nannochloropsis oculate and Chlorella vulgaris for biodiesel production. Chem Eng Process Process Intensif 48(6):1146–1151

    Article  CAS  Google Scholar 

  • Da Ros PCM, Silva CSP, Silva-Stenico ME, Fiore MF, De Castro HF (2013) Assessment of chemical and physicochemical properties of cyanobacterial lipids for biodiesel production. Mar Drugs 11(7):2365–2381

    Article  Google Scholar 

  • Dayananda C, Sarada R, Usha Rani M, Shamala TR, Ravishankar GA (2007) Autotrophic cultivation of Botryococcus braunii for the production of hydrocarbons and exopolysaccharides in various media. Biomass Bioenergy 31:87–93

    Article  CAS  Google Scholar 

  • Demirbas A (1998) Fuel properties and calculation of higher heating values of vegetable oils. Fuel 77(9/10):1117–1120

    Article  CAS  Google Scholar 

  • DuBois M, Gilles KA., Hamilton JK., Rebers PA, Smith F (1956) Colorimetric Method for Determination of Sugars and Related Substances. Anal Chem 28(3):350–356

  • El-Shimi HI et al. (2013) Biodiesel production from Spirulina-platensis microalgae by in-situ transesterification process. Journal of Sustainable Bioenergy Systems 3(3):224

  • Fan J, Yan C, Andre C, Shanklin J, Schwender J, Xu C (2012) Oil accumulation is controlled by carbon precursor supply for fatty acid synthesis in Chlamydomonas reinhardtii. Plant Cell Physiol 53:1380–1390. https://doi.org/10.1093/pcp/pcs082

    Article  CAS  Google Scholar 

  • Fernandes B, Teixeira J, Dragone G, Vicente AA, Kawano S, Bišová K, Přibyl P, Zachleder V, Vítová M (2013) Relationship between starch and lipid accumulation induced by nutrient depletion and replenishment in the microalga Parachlorella kessleri. Bioresour Technol 144:268–274

    Article  CAS  Google Scholar 

  • Francisco EC, Neves DB, Jacob-Lopes E, Franco TT (2010) Microalgae as feedstock for biodiesel production: carbon dioxide sequestration, lipid production and biofuel quality. J Chem Technol Biotechnol 85:395–403

    Article  CAS  Google Scholar 

  • Franco-Navarro JD, Brumós J, Rosales MA, Cubero-Font P, Talón M, and Colmenero-Flores JM (2015) Chloride regulates leaf cell size and water relations in tobacco plants. J Exp Bot 67(3):873-891.

  • Frankel EN (1998) Lipid oxidation. Dundee: The Oily Press. Vol 10

  • Fung KS, Liew EWT, Ngu HLN (2013) Optimization of nutrient media composition for microalgae biomass production using central composite design. Chemeca 2013: Challenging Tomorrow, p.278

  • Ge Y, Liu J, Tian G (2011) Growth characteristics of Botryococcus braunii 765 under high CO2 concentration in photobioreactor. Bioresour Technol 102(1):130–134

    Article  CAS  Google Scholar 

  • Gopinath A, Puhan S, Nagarajan G (2009) Relating the cetane number of biodiesel fuels to their fatty acid composition: a critical study. Proc Inst Mech Eng D: Journal of Automobile Engineering 223(4):565–583

    Article  Google Scholar 

  • Gorain PC, Bagchi SK, Mallick N (2013) Effects of calcium, magnesium and sodium chloride in enhancing lipid accumulation in two green microalgae. Environ Technol 34(13–14):1887–1894

    Article  CAS  Google Scholar 

  • Gorman DS, Levine RP (1965) Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardi. Proc Natl Acad Sci U S A 54:1665–1669

    Article  CAS  Google Scholar 

  • Gouveia L, Oliveira AC (2009) Microalgae as a raw material for biofuels production. J Ind Microbiol Biotechnol 36(2):269–274

    Article  CAS  Google Scholar 

  • Hakkalin NLS, Paz AP, Aranda DAG, Moraes LMP (2014) Enhancement of cell growth and lipid content of a freshwater microalga Scenedesmus sp. by optimizing nitrogen, phosphorus and vitamin concentrations for biodiesel production. Nat Sci 6:1044–1054

    Google Scholar 

  • Held P, Raymond K, (2011) Determination of algal cell lipids using Nile red—using microplates to monitor neutral lipids in Chlorella vulgaris

  • Ho SH, Chen CY, Chang JS (2012) Effect of light intensity and nitrogen starvation on CO2 fixation and lipid/carbohydrate production of an indigenous microalga Scenedesmus obliquus CNW-N. Bioresour Technol 113:244–252

    Article  CAS  Google Scholar 

  • Hu Q (2004) Environmental effects on cell composition. In: Richmond, A. (Ed.), Handbook of Microalgal Culture: Biotechnology and Applied Phycology. Blackwell Science, Victoria, pp. 83–93

  • Hu Q, Sommerfeld M, Jarvis E, Ghirardi M, Posewitz M, Seibert M, Darzins A (2008) Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. Plant J 54:621–639. https://doi.org/10.1111/j.1365-313X.2008.03492

    Article  CAS  Google Scholar 

  • Kapdan IK, Aslan S (2008) Application of the Stover–Kincannon kinetic model to nitrogen removal by Chlorella vulgaris in a continuously operated immobilized photobioreactor system. J Chem Technol Biotechnol 83:998–1005

    Article  CAS  Google Scholar 

  • Knothe G (2002) Structure indices in FA chemistry: how relevant is the iodine value? J Am Oil Chem Soc 79:847–854

    Article  CAS  Google Scholar 

  • Knothe GH (2005) Dependence of biodiesel fuel properties on the structure of fatty acid alkyl esters. Fuel Process Technol 86:1059–1070

    Article  CAS  Google Scholar 

  • Knothe G (2006) Analyzing biodiesel: standards and other methods. J Am Oil Chem Soc 83:823–833

    Article  CAS  Google Scholar 

  • Knothe G (2008) “Designer” biodiesel: optimizing fatty ester composition to improve fuel properties. Energy Fuel 22:1358–1364

    Article  CAS  Google Scholar 

  • Kona R, Hemlatha M, Srivastav KV, Mohan SV (2017) Regulatory effect of Fe-EDTA on mixotrophic cultivation of Chlorella sp. towards biomass growth and metabolite production. Bioresour Technol 244:1227–1234

    Article  CAS  Google Scholar 

  • Krisnangkura KA (1986) A simple method for estimation of cetane index of vegetable oil methyl esters. J Am Oil Chem Soc 63:552–553

    Article  CAS  Google Scholar 

  • Ladommatos N, Parsi M, Knowles A (1996) The effect of fuel cetane improver on diesel pollutant emissions. Fuel 75(1):8–14

    Article  CAS  Google Scholar 

  • Li Y, Han D, Hua G, Dauvillee D, Sommerfeld M, Ball S, Hu Q (2010) Chlamydomonas starchless mutant defective in ADP-glucose pyrophosphorylase hyper-accumulates triacylglycerols. Metab Eng 12:387–391. https://doi.org/10.1016/j.ymben.2010.02.002

    Article  CAS  Google Scholar 

  • Li Y, Han D, Sommerfeld M, Hu Q (2011) Photosynthetic carbon partitioning and lipid production in the oleaginous microalga Pseudochlorococcum sp. (Chlorophyceae) under nitrogen-limited conditions. Bioresour Technol 102:123–129. https://doi.org/10.1016/j.biortech.2010.06.03

    Article  CAS  Google Scholar 

  • Liu X, Sheng J, Curtiss R III (2011) Fatty acid production in genetically modified cyanobacteria. Proc Natl Acad Sci U S A 108:6899–6904. https://doi.org/10.1073/pnas.1103014108

    Article  Google Scholar 

  • Los DA, Murata N (2004) Membrane fluidity and its roles in the perception of environmental signals. Biochim Biophys Acta Biomembr 1666(1):142–157

    Article  CAS  Google Scholar 

  • Lv JM, Cheng LH, Xu XH, Zhang L, Chen HL (2010) Enhanced lipid production of Chlorella vulgaris by adjustment of cultivation conditions. Bioresour Technol 101(17):6797–6804

    Article  CAS  Google Scholar 

  • Martin W, Scheibe R, Schnarenberger C (2000) The Calvin cycle and its regulation. In: Leegood RC, C R, Shakey TD, von Caemmerer S (eds) Photosynthesis: Physiology and Metabolism. Kluwer Academic Publishers, Dordrecht, pp 9–51

    Chapter  Google Scholar 

  • Mayers JJ, Flynn KJ, Shields RJ (2014) Influence of the N:P supply ratio on biomass productivity and time-resolved changes in elemental and bulk biochemical composition of Nannochloropsis sp. Bioresour Technol 169:588–595

    Article  CAS  Google Scholar 

  • Nelson DL, Cox MM (2008) Lipid biosynthesis. In: Principles of biochemistry, 4th edn. W. H. Freeman and Company, New York, pp 805–845

    Google Scholar 

  • Nichols HW, Bold HC (1965) Trichosarcina polymorpha gen. et sp. nov. J Phycol 1(1):34–38

    Article  Google Scholar 

  • Oijen VT, Leeuwe VMA, Gieskes WW, de Baar HJ (2004) Effects of iron limitation on photosynthesis and carbohydrate metabolism in the Antarctic diatom Chaetoceros brevis (Bacillariophyceae). Eur J Phycol 39(2):161–171

    Article  CAS  Google Scholar 

  • Radakovits R, Jinkerson RE, Darzins A, Posewitz MC (2010) Genetic engineering of algae for enhanced biofuel production. Eukaryot Cell 9:486–501

    Article  CAS  Google Scholar 

  • Rakesh S, Dhar DW, Prasanna R, Saxena AK, Saha S, Shukla M, Sharma K (2015) Cell disruption methods for improving lipid extraction efficiency in unicellular microalgae. Eng Life Sci 15(4):443–447. https://doi.org/10.1002/elsc.201400222

    Article  CAS  Google Scholar 

  • Ramírez-Verduzco LF, Rodriguez-Rodriguez JE, Jaramillo-Jacob AR (2012) Predicting cetane number, kinematic viscosity, density and higher heating value of biodiesel from its fatty acid methyl ester composition. Fuel 91:102–111

    Article  CAS  Google Scholar 

  • Ramos MJ, Fernández CM, Casas A, Rodríguez L, Pérez Á (2009) Influence of fatty acid composition of raw materials on biodiesel properties. Bioresour Technol 100(1):261–268

    Article  CAS  Google Scholar 

  • Raven JA (2016) Chloride: essential micronutrient and multifunctional beneficial ion. J Exp bot 68(3):359–367.

  • Ruangsomboon S (2015) Effects of different media and nitrogen sources and levels on growth and lipid of green microalga Botryococcus braunii KMITL and its biodiesel properties based on fatty acid composition. Bioresour technol 191:377–384

  • Ruangsomboon S, Ganmanee M, Choochote S (2013) Effects of different nitrogen, phosphorus, and iron concentrations and salinity on lipid production in newly isolated strain of the tropical green microalga, Scenedesmus dimorphus KMITL. J Appl Phycol 25:867–874

    Article  CAS  Google Scholar 

  • Saraf S, Thomas B (2007) Influence of feedstock and process chemistry on biodiesel quality. Process Saf Environ Prot 85:360–364

    Article  CAS  Google Scholar 

  • Sarin A, Arora R, Singh NP, Sarin R, Malhotra RK, Kundu K (2009) Effect of blends of palm-jatropha-pongamia biodiesels on cloud point and pour point. Energy 34:2016–2021

    Article  CAS  Google Scholar 

  • Sarpal AS, Costa ICR, Teixeira CMLL, Filocomo D, Candido R et al (2016) Investigation of biodiesel potential of biomasses of Microalgaes chlorella, spirulina and tetraselmis by NMR and GC-MS techniques. J Biotechnol Biomater 6:220

    Google Scholar 

  • Sharma KK, Schuhmann H, Schenk PM (2012) High lipid induction in microalgae for biodiesel production. Energies 5:1532–1553

    Article  CAS  Google Scholar 

  • Siaut M, Cuine S, Cagnon C, Fessler B, Naguyen M, Carrier P, Beyly A, Beisson F, Triantaphylidès C, Li-Beisson Y, Gilles P (2011) Oil accumulation in the model green alga Chlamydomonas reinhardtii: characterization, variability between common laboratory strains and relationship with starch reserves. BMC Biotechnol 11:7

    Article  CAS  Google Scholar 

  • Song M, Pei H, Hu W, Ma G (2013) Evaluation of the potential of ten microalgal strains for biodiesel production. Bioresour Technol 141:245–251

    Article  CAS  Google Scholar 

  • Sorokin C (1959) Tabular comparative data for the low-and high-temperature strains of chlorella. Nature 184:613–614

    Article  CAS  Google Scholar 

  • Stanier RY, Kunisawa R, Mandel M, Cohen-Bazire G (1971) Purification and properties of unicellular blue-green algae (order Chroococcales). Bacteriol Rev 35:171–205

    CAS  Google Scholar 

  • Tetlow IJ, Morell MK, Emes MJ (2004) Recent developments in understanding the regulation of starch metabolism in higher plants. J Exp Bot 55:2131–2145

    Article  CAS  Google Scholar 

  • Work VH, Radakovits R, Jinkerson RE, Meuser JE, Elliott LG, Vinyard DJ, Laurens LML, Dismukes GC, Posewitz M (2010) Increased lipid accumulation in the Chlamydomonas reinhardtii sta7-10 starchless isoamylase mutant and increased carbohydrate synthesis in complemented strain. Eukaryot Cell 9:1251–1269. https://doi.org/10.1128/EC.00075-10

    Article  CAS  Google Scholar 

  • Yeh KL, Chang JS (2012) Effects of cultivation conditions and media composition on cell growth and lipid productivity of indigenous microalga Chlorella vulgaris ESP-31. Bioresour Technol 105:120–127

    Article  CAS  Google Scholar 

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Acknowledgements

The authors are grateful to Director, IARI, New Delhi, for essential facilities.

Funding

The study is financially supported by the Department of Biotechnology, Govt. of India under Indo-Denmark Collaborative Project.

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Authors and Affiliations

  1. Centre for Conservation and Utilisation of Blue Green Algae, ICAR-Indian Agricultural Research Institute, New Delhi, 110012, India

    Rashi Vishwakarma, Dolly Wattal Dhar & Sudhir Saxena

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  1. Rashi Vishwakarma
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  2. Dolly Wattal Dhar
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  3. Sudhir Saxena
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Correspondence to Rashi Vishwakarma.

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Vishwakarma, R., Dhar, D.W. & Saxena, S. Influence of nutrient formulations on growth, lipid yield, carbon partitioning and biodiesel quality potential of Botryococcus sp. and Chlorella sp.. Environ Sci Pollut Res 26, 7589–7600 (2019). https://doi.org/10.1007/s11356-019-04213-2

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Keywords

  • Lipid
  • Nutrient media
  • Biodiesel
  • FAMEs
  • Biomass
  • Microalgae