Use of a Combined Technology of Ultrasonication, Three-Phase Partitioning, and Aqueous Enzymatic Oil Extraction for the Extraction of Oil from Spirogyra sp.
Algal oil from Spirogyra sp. was extracted using a combined technology of ultrasonication, three-phase partitioning, and aqueous enzymatic oil extraction. Ultrasonication was done to rupture the cell wall and papain was used for an easier release of the trapped oil. The salt concentration for three-phase partitioning, preincubation period with (or without) the protease, and its operational temperature were optimized for a maximum possible yield of the oil and the effect of ultrasonication, and three-phase partitioning with (or without) the protease were studied. It was found that under optimized conditions at 50% ammonium sulphate concentration using tert-butanol (in 1？:？1, v/v ratio) a presonicated and papain treated algal suspension could produce 24% (w/w, dry weight) oil within few hours which was ten times higher as compared to the oil obtained by Soxhlet extraction using hexane and two times higher than the oil obtained without using the protease. 1. Introduction In past few decades preparation of biofuels from the waste materials has been a matter of great interest [1–3]. A more recent alternative which has been in focus for acting as a source of oil is algae [1, 2, 4]. The algae which have been found most effective as triacylglycerols (TAG) source are microalgae. Depending upon the species and the extraction process, the TAG content (%, dry weight of the algae) in microalgae can reach beyond 90% which is much higher than the reported values (20–30%) for macroalgae [5–7]. However, extraction of oil from macroalgae has continued to be a matter of research [4, 5, 8]. In the present work Spirogyra sp. has been evaluated as the source of oil. Apart from its widespread availability one attractive feature of Spirogyra cell is its simple cell wall which can be easily ruptured [9]. Hence the algae need more attention and should be explored with advanced technologies which speed up the extraction process and improve the yield of the oil. One of the recent techniques which has been attractive to the scientists for oil extraction is the use of Three-Phase Partitioning (TPP) [10–12]. Three-Phase Partitioning (TPP) uses tert-butanol and ammonium sulfate to precipitate enzymes and proteins from aqueous solutions. TPP can be used effectively with crude samples and can be easily scaled up. tert-Butanol seems to bind with TPP-precipitated proteins which are actually protein-tert-butanol coprecipitates float in between organic and aqueous layer. Buoyancy of this protein bound tert-butanol above the denser saline aqueous medium and presence of sulphate ion in large
References
[1]
E. G. Shay, “Diesel fuel from vegetable oils: status and opportunities,” Biomass and Bioenergy, vol. 4, no. 4, pp. 227–242, 1993.
[2]
J. W. Lee, Advanced Biofuels and Bioproducts, Springer, NewYork, NY, USA, 2013.
[3]
R. Kasirajan, S. Pandian, S. Tamilarasan, and R. Sahadevan, “Lipid extraction from natural plant source of Adenanthera pavonina using mixed solvent by superheated extractor,” Korean Journal of Chemical Engineering, vol. 31, no. 3, pp. 509–513, 2014.
[4]
T. Suganya, R. Kasirajan, and S. Renganathan, “Ultrasound-enhanced rapid in situ transesterification of marine macroalgae Enteromorpha compressa for biodiesel production,” Bioresource Technology, vol. 156, pp. 283–290, 2014.
[5]
A. B. M. S. Hossain, A. Salleh, A. N. Boyce, P. Chowdhury, and M. Naqiuddin, “Biodiesel fuel production from algae as renewable energy,” American Journal of Biochemistry and Biotechnology, vol. 4, no. 3, pp. 250–254, 2008.
[6]
E. Ryckebosch, K. Muylaert, and I. Foubert, “Optimization of an analytical procedure for extraction of lipids from microalgae,” Journal of the American Oil Chemists' Society, vol. 89, no. 2, pp. 189–198, 2012.
[7]
P. Mercer and R. E. Armenta, “Developments in oil extraction from microalgae,” European Journal of Lipid Science and Technology, vol. 113, no. 5, pp. 539–547, 2011.
[8]
S. Gupta, R. Sharma, S. K. Soni, and S. Sharma, “Biomass utilization of waste algal consortium for extraction of algal oil,” Journal of Algal Biomass Utilization, vol. 3, pp. 34–38, 2012.
[9]
F. S. Eshaq, M. N. Ali, and M. K. Mohd, “Spirogyra biomass a renewable source for biofuel (bioethanol) production,” International Journal of Engineering Science and Technology, vol. 2, pp. 7045–7054, 2010.
[10]
A. Sharma, S. K. Khare, and M. N. Gupta, “Three phase partitioning for extraction of oil from soybean,” Bioresource Technology, vol. 85, no. 3, pp. 327–329, 2002.
[11]
A. Sharma and M. N. Gupta, “Oil extraction from almond, apricot and rice bran by three-phase partitioning after ultrasonication,” European Journal of Lipid Science and Technology, vol. 106, no. 3, pp. 183–186, 2004.
[12]
S. Shah, A. Sharma, and M. N. Gupta, “Extraction of oil from Jatropha curcas L. seed kernels by enzyme assisted three phase partitioning,” Industrial Crops and Products, vol. 20, no. 3, pp. 275–279, 2004.
[13]
C. Dennison and R. Lovrien, “Three phase partitioning: concentration and purification of proteins,” Protein Expression and Purification, vol. 11, no. 2, pp. 149–161, 1997.
[14]
S. Raghava, B. Barua, P. K. Singh et al., “Refolding and simultaneous purification by three-phase partitioning of recombinant proteins from inclusion bodies,” Protein Science, vol. 17, no. 11, pp. 1987–1997, 2008.
[15]
A. Rosenthal, D. L. Pyle, and K. Niranjan, “Aqueous and enzymatic processes for edible oil extraction,” Enzyme and Microbial Technology, vol. 19, no. 6, pp. 402–420, 1996.
[16]
A. Sharma, S. K. Khare, and M. N. Gupta, “Enzyme-assisted aqueous extraction of peanut oil,” Journal of the American Oil Chemists' Society, vol. 79, no. 3, pp. 215–218, 2002.
[17]
S. Shah, A. Sharma, and M. N. Gupta, “Extraction of oil from Jatropha curcas L. seed kernels by combination of ultrasonication and aqueous enzymatic oil extraction,” Bioresource Technology, vol. 96, no. 1, pp. 121–123, 2005.
[18]
L. C. Dickey, D. B. Johnston, M. J. Kurantz, A. McAloon, and R. A. Moreau, “Modification of aqueous enzymatic oil extraction to increase the yield of corn oil from dry fractionated corn germ,” Industrial Crops and Products, vol. 34, no. 1, pp. 845–850, 2011.
[19]
I. Horst, B. M. Parker, J. S. Dennis, C. J. Howe, S. A. Scott, and A. G. Smith, “Treatment of Phaeodactylum tricornutum cells with papain facilitates lipid extraction,” Journal of Biotechnology, vol. 162, no. 1, pp. 40–49, 2012.
[20]
N. L. Huang, M.-D. Huang, T.-L. Chen, and A. H. C. Huang, “Oleosin of subcellular lipid droplets evolved in green algae,” Plant Physiology, vol. 161, no. 4, pp. 1862–1874, 2013.
[21]
B. S. Baines and K. Brocklehurst, “A necessary modification to the preparation of papain from any high-quality latex of Carica papaya and evidence for the structural integrity of the enzyme produced by traditional methods,” Biochemical Journal, vol. 177, no. 2, pp. 541–548, 1979.
[22]
E. M. Frenkel, “Studies on enzyme action: XV. Factors influencing the proteolytic activity of papain,” The Journal of Biological Chemistry, vol. 31, pp. 201–215, 1917.
[23]
H. Bisswanger, Practical Enzymology, Wiley-VCH Verlag Gmbh, Weinheim, Germany, 2004.
