Glycerol on Lipid Enhancement and FAME Characterization in Algae for Raw Material of Biodiesel

Anwesha Khanra, Shrasti Vasistha, Monika Prakash Rai


To reduce the cost of media components and consecutively enhance the amount of lipid formation, the effect of crude glycerol on Euglena gracilis is reported for the first time. Presently, E. gracilis has been chosen because of excellent capability of its growth and lipid synthesis in mixotrophic culture condition on organic carbon sources (Glucose and Glycerol). Biochemical composition of the cell in presence of both organic carbon sources were compared in mixotrophic condition. Glycerol was selected as most suitable carbon source and the highest biomass concentration of 2.63 g L-1 and lipid accumulation of 27.64 % were observed. The effect of biodiesel derived crude glycerol on the growth of E. gracilis was found very positive. A significant increase in lipid accumulation (49.46%) was noticed in presence of crude glycerol by optimizing concentration and other culture parameters. Fatty acid methyl esters (FAMEs) produced from lipid biomolecules by following transesterification reaction, were analysed through GC-MS. FAMEs governed by 93.458% C16-C18 fatty acids with appropriate quantities of Saturated fatty acids (SFA) and Unsaturated fatty acids (UFA). Several requisite fuel properties were estimated and found to be in accordance with American and European biodiesel standards. Hence, this study focuses the improved lipid synthesis in E. gracilis utilizing crude glycerol for the preparation of raw biodiesel and it supports biorefinery approach.

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Algae, biomass energy, Mixotrophy

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R. Tripathi, J. Singh, I.S. Thakur, “Characterization of microalga Scenedesmus sp. ISTGA1 for potential CO2 sequestration and biodiesel production”, Renew Energy, 74 (2015), pp.774-781.

A. Ebrahimian, H.R. Kariminia, M. Vosoughi, “Lipid production in mixotrophic cultivation of Chlorella vulgaris in a mixture of primary and secondary municipal wastewater”, Renew Energy, 71,(2014), pp. 502-508.

A.L. Goncalves, M. Simoes, J.C.M. Pires, “The effect of light supply on microalgal growth, CO2 uptake and nutrient removal from wastewater”, Energ Convers Manage, 85 (2014), pp. 530-536.

B. Kiran, R. Kumar, D. Desmukh, “Perspectives of microalgal biofuels as a renewable source of energy”, Energ Converse Manage., 88 (2014), pp. 1228-44.

S. Vidyashankar, V.S.K. Gopal, G.V. Swarnalatha, M.D. Kavitha, V.S. Chauhan, R. Ravi, A.K. Bansal, R. Singh, A. Pande, G.A. Ravishankar, R. Sarada, “Characterization of fatty acids and hydrocarbons of chlorophycean microalgae towards their use as biofuel source”, Biomass and Bioenerg., 77 (2015), pp. 75-91.

E.C. Franscisco, D.B. Neves, L. Jocob, E. Fanco, “Microalgae as feed stocks for biodiesel production: CO2 sequestration, lipid production and biofuel quality”, J Chem Technol Biot, 85 (2010), pp. 395-403.

R. Karpagam, K.J. Raj, Ashok Kumar B, P. Varalakshmi, “Characterization and fatty acid profiling in two fresh water microalgae for biodiesel production: Lipid enhancement methods and media optimization using response surface methodology”, Bioresour Technol., 188 (2015), pp. 177-184.

R. Valez-ojeda, M. Gonzalez- Munoz, R. Vs-Vazquez, J.N. Zapat, J.C. Chavarria- herandez, S.L. Adrian, F.B. Perez, T.T. Thompson, G.G. Solorzano, R.M.E.G. Medrano, “Characterization of five fresh water microalgae with potential for biodiesel production”, Algal Res, 7 (2015), pp. 33-44.

Y. Liang, N. Sarkany, Y. Cui, “Biomass and lipid productivities of Chlorella vulgaris under autotrophic, heterotrophic and mixotrophic growth conditions”, Biotechnol Lett, 31 (2009), pp. 1043-1049.

M.P. Rai, S. Nigam, R. Sharma, “Response of growth and fatty acid compositions of Chlorella pyrenoidosa under mixotrophic cultivation with acetate and glycerol for bioenergy application”, Biomass and Bioenerg, 58 (2013), pp. 251-257.

S. Kim, J.E. Park, Y.B. Cho, S.J. Hwang, “Growth rate, organic carbon and nutrient removal rates of Chlorella sorokiniana in autotrophic, heterotrophic and mixotrophic conditions”, Bioresour Technol, 144 (2013), pp. 8-13.

I. Pancha, K. Chokshi, S. Mishra, “Enhanced biofuel production potential with nutritional stress amelioration through optimization of carbon source and light intensity in Scenedesmus sp. CCNM 1077”, Bioresour Technol., 179 (2015), pp. 565-572.

H.J. Choi, S.W. Yu, “Influence of crude glycerol on the biomass and lipid content of microalgae”, Biotechnol Biotechnol Equip., 29(4) (2015), pp. 506-513.

J. Trivedi, M. Aila, D.P. Bangwal, S. Kaul, M.O. Garg, “Algae based biorefinery- How to make sense?” Renew Sust Energ Rev., 47 (2015), pp. 295-307.

B. Cheirsilp, S. Torpee, “Enhanced growth and lipid production of microalgae under mixotrophic culture condition: Effect of light intensity, glucose concentration and fed-batch cultivation”, Bioresour Technol., 110 (2012), pp. 510-516.

G. Knothe, “Improving biodiesel fuel properties by modifying fatty ester composition”. Ener Environ Sci. 2 (2009), pp. 759-66.

G. Knothe, “Production and properties of biodiesel from algal oils. In: Borowitzka MA, Moheimani NR (eds),” Algae for biofuels and energy. Springer 2013, pp. 207-221.

P. Neofotis, A. Huang, K. Sury, W. Chang, F. Joseph, A. Gabr, S. Twary, W. Qiu, O. Holguin, E.W.J. Polle, “Characterization and classification of highly productive microalgae strains discovered for biofuel and bioproduct generation”, Algal Res, 15 (2016), pp.164-178.

T. Ogawa, M. Tamoi, A. Kimura, A. Mine, H. Sakuyama, E. Yoshida, T. Maruta, K. Suzuki, T. Ishikawa, S. Shigeoka, “Enhancement of photosynthetic capacity in Euglena gracilis by expression of cyanobacterial fructose-1,6-/sedoheptulose-1,7-bisphosphatase leads to increases in biomass and wax ester production”, Biotechnol Biofuels, 8 (2015), pp. 80-91.

Y. Yamane, T. Utsunomiya, M. Watanabe, K. Sasaki, “Biomass production in mixotrophic culture of Euglena gracilis under acidic condition and its growth energetics”, Biotechnol Lett., 23 (2011), pp. 1223-1228.

V.T. Ramachandra, N.H. Chanakya, M.D. Mahapatra, “Euglena sp. as a suitable source of lipids for potential use as biofuel and sustainable wastewater treatment”, J Appl Phycol, 25(3), (2013), pp. 855-865.

M.P. Rai, S. Gupta, “Effect of media composition and light supply on biomass, lipid content and FAME profile for quality biofuel production from Scenedesmus abundans”, Energy Converse Manage, 2016, (online available May 2016).

O.H. Lowry, N.J. Rosebrough, A.L. Farr, R.J. Randall, “Protein measurement with the folin phenol reagent”, J Biol Chem 1951(193), pp. 265-275.

X. Miao, Q. Wu. “High yield bio-oil production from fast pyrolysis by metabolic controlling of Chlorella protothecoides,” J Biotechnol.110(1), (2004), pp. 85-93.

S. Leow, R.J. Witter, R.D. Vardon, K.B Sharma., S.J. Guest, J.T. Sarathmann, “Prediction of microalgae hydrothermal liquifection products from feedstock biochemical composition”, Green Chemistry, 17 (2015), pp. 3584-3599.

Anders Moller, Danish Food information 2011, Sheppard A, J Lipid Manual, U.S. Food and Drug Administration 1992.

G. Yu, D. Shi, Z. Cai, W. Cong, F. Ouyang, “Growth and physiological features of Cyanobacterium Anabaena sp. strain PCC 7120 in a glucose-mixotrophic culture”, Chinese J Chem Eng., 19(2011), pp. 108-115.

P. Nicolas, G. Freyssinet, V. Nigon, “Effect of light on glucose utilization by Euglena gracilis,” Plant Physiol, 65 (1980), pp. 631-634.

J.P. Schwarzhans, D. Cholewa, P. Grimn, U. Beshay, J.M. Risse, K. Friehs, E. Flaschel, “Dependency of the fatty acid composition of Euglena gracilis on growth phase and culture conditions”, J Appl Phycol, 27(4), (2014), pp. 1389-1399.

V. Andruleviciute, V. Makareviciene, V. Skorupskaite, M. Gumbyte, “Biomass and oil content of Chlorella sp., Haematococcus sp., Nannochloris sp. and Scenedesmus sp. under mixotrophic growth conditions in the presence of technical glycerol,” J Appl Phycol., 26 (2014), pp. 83-90.

G. Knothe, “Fuel properties of highly polyunsaturated fatty acid methyl esters. Prediction of fuel properties of algal biodiesel”, Ener Fuel 26 (2012), pp. 5265-5273.

M.J. Ramos, C.M. Fernandez, A. Casas, L. Rodriguez, A. Perez, “Influence of fatty acid composition of raw materials on biodiesel properties”, Bioresour Technol, 100 (2009), pp. 261-268.


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