Cite this paper:
CHIA Shir Reen, CHEW Kit Wayne, SHOW Pau Loke, SIVAKUMAR Manickam, LING Tau Chuan, TAO Yang. Isolation of protein from Chlorella sorokiniana CY1 using liquid biphasic flotation assisted with sonication through sugaring-out effect[J]. Journal of Oceanology and Limnology, 2019, 37(3): 898-908

Isolation of protein from Chlorella sorokiniana CY1 using liquid biphasic flotation assisted with sonication through sugaring-out effect

CHIA Shir Reen1,2, CHEW Kit Wayne1,2, SHOW Pau Loke1,2, SIVAKUMAR Manickam2, LING Tau Chuan3, TAO Yang4
1 Bioseparation Research Group, Department of Chemical and Environmental Engineering, Faculty of Engineering, University of Nottingham Malaysia Campus, Semenyih 43500, Malaysia;
2 Department of Chemical and Environmental Engineering, Faculty of Engineering, University of Nottingham Malaysia Campus, Semenyih 43500, Malaysia;
3 Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur 50603, Malaysia;
4 College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Abstract:
Microalgae, a sustainable source of multi beneficial components has been discovered and could be utilised in pharmaceutical, bioenergy and food applications. This study aims to investigate the sugaring-out effect on the recovery of protein from wet green microalga, Chlorella sorokiniana CY 1 which was assisted with sonication. A comparison of monosaccharides and disaccharides as one of the phaseforming constituents shows that the monosaccharides, glucose was the most suitable sugar in forming the phases with acetonitrile to enhance the production of protein (52% of protein). The protein productivity of microalgae was found to be significantly influenced by the volume ratio of both phases, as the yield of protein increased to 77%. The interval time between the sonication as well as the sonication modes were influencing the protein productivity as well. The optimum protein productivity was obtained with 10 s of resting time in between sonication. Pulse mode of sonication was suitable to break down the cell wall of microalgae compared to continuous mode as a lower protein yield was obtained with the application of continuous mode. The optimum condition for protein extraction were found as followed:200 g/L glucose as bottom phase with volume ratio of 1:1.25, 10 s of resting time for ultrasonication, 5 s of ultrasonication in pulse mode and 0.25 g of biomass weight. The high yield of protein about 81% could be obtained from microalgae which demonstrates the potential of this source and expected to play an important role in the future.
Key words:    sugaring-out|Chlorella sorokiniana CY1|liquid biphasic flotation|sonication|microalgae|extraction   
Received: 2018-09-13   Revised: 2018-10-19
Tools
PDF (838 KB) Free
Print this page
Add to favorites
Email this article to others
Authors
Articles by CHIA Shir Reen
Articles by CHEW Kit Wayne
Articles by SHOW Pau Loke
Articles by SIVAKUMAR Manickam
Articles by LING Tau Chuan
Articles by TAO Yang
References:
Asenjo J A, Andrews B A. 2012. Aqueous two-phase systems for protein separation:phase separation and applications.Journal of Chromatography A, 1238:1-10, https://doi.org/10.1016/j.chroma.2012.03.049.
Becker E W. 2007. Micro-algae as a source of protein.Biotechnology Advances, 25(2):207-210.
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, https://doi.org/10.1016/0003-2697(76)90527-3.
Cardoso G D B, Mourão T, Pereira F M, Freire M G, Fricks A T, Soares C M F, Lima Á S. 2013. Aqueous two-phase systems based on acetonitrile and carbohydrates and their application to the extraction of vanillin. Separation and Purification Technology, 104:106-113, https://doi.org/10.1016/j.seppur.2012.11.001.
Chemat F, Rombaut N, Meullemiestre A, Turk M, Perino S, Fabiano-Tixier A S, Abert-Vian M. 2017a. Review of green food processing techniques. Preservation, transformation, and extraction. Innovative Food Science& Emerging Technologies, 41:357-377, https://doi.org/10.1016/j.ifset.2017.04.016.
Chemat F, Rombaut N, Sicaire A G, Meullemiestre A, FabianoTixier A S, Abert-Vian M. 2017b. Ultrasound assisted extraction of food and natural products. Mechanisms, techniques, combinations, protocols and applications. A review. Ultrasonics Sonochemistry, 34:540-560, https://doi.org/10.1016/j.ultsonch.2016.06.035.
Chew K W, Chia S R, Show P L, Ling T C, Arya S S, Chang J S. 2018. Food waste compost as an organic nutrient source for the cultivation of Chlorella vulgaris. Bioresource Technology, 267:356-362, https://doi.org/10.1016/j.biortech.2018.07.069.
Chew K W, Yap J Y, Show P L, Suan N H, Juan J C, Ling T C, Lee D J, Chang J S. 2017. Microalgae biorefinery:high value products perspectives. Bioresource Technology, 229:53-62, https://doi.org/10.1016/j.biortech.2017.01. 06.
Chia S R, Chew K W, Show P L, Yap Y J, Ong H C, Ling T C, Chang J S. 2018b. Analysis of economic and environmental aspects of microalgae biorefinery for biofuels production:a review. Biotechnology Journal, 13(6):1 700 618, https://doi.org/10.1002/biot.201700618.
Chia S R, Ong H C, Chew K W, Show P L, Phang S M, Ling T C, Nagarajan D, Lee D J, Chang J S. 2018a. Sustainable approaches for algae utilisation in bioenergy production.Renewable Energy, 129:838-852, https://doi.org/10.1016/j.renene.2017.04.001.
Dhamole P B, Mahajan P, Feng H. 2010a. Sugaring out:a new method for removal of acetonitrile from preparative RPHPLC eluent for protein purification. Process Biochemistry, 45(10):1 672-1 676, https://doi.org/10.1016/j.procbio.2010.06.020.
Dhamole P B, Mahajan P, Feng H. 2010b. Phase separation conditions for sugaring-out in acetonitrile-water systems.J. Chem. Eng. Data, 55(9):3 803-3 806, https://doi.org/10.1021/je1003115.
Frampton J P, Tsuei M, White J B, Abraham A T, Takayama S. 2015. Aqueous two-phase system-mediated antibody micropatterning enables multiplexed immunostaining of cell monolayers and tissues. Biotechnology Journal, 10(1):121-125, https://doi.org/10.1002/biot.201400271.
Gerken H G, Donohoe B, Knoshaug E P. 2013. Enzymatic cell wall degradation of Chlorella vulgaris and other microalgae for biofuels production. Planta, 237(1):239-253, https://doi.org/10.1007/s00425-012-1765-0.
Goja A M, Yang H, Cui M, Li C. 2013. Aqueous two-phase extraction advances for bioseparation. Journal of Bioprocessing & Biotechniques, 4:140, https://doi.org/10.4172/2155-9821.1000140.
Guo S J, Li J, Li T, Shi D Y, Han L J. 2011. Synthesis of three bromophenols from red algae as PTP1B inhibitors.
Chinese Journal of Oceanology and Limnology, 29(1):68-74, https://doi.org/10.1007/s00343-011-9996-7.
Hariskos I, Posten C. 2014. Biorefinery of microalgaeopportunities and constraints for different production scenarios. Biotechnology Journal, 9(6):739-752, https://doi.org/10.1002/biot.201300142.
Ho S H, Chen Y D, Chang C Y, Lai Y Y, Chen C Y, Kondo A, Ren N Q, Chang J S. 2017. Feasibility of CO2 mitigation and carbohydrate production by microalga Scenedesmus obliquus CNW-N used for bioethanol fermentation under outdoor conditions:effects of seasonal changes.Biotechnology for Biofuels, 10:27, https://doi.org/10.1186/s13068-017-0712-5.
Khadhraoui B, Turk M, Fabiano-Tixier A S, Petitcolas E, Robinet P, Imbert R, El Maâtaoui M, Chemat F. 2018.Histo-cytochemistry and scanning electron microscopy for studying spatial and temporal extraction of metabolites induced by ultrasound. Towards chain detexturation mechanism. Ultrasonics Sonochemistry, 42:482-492, https://doi.org/10.1016/j.ultsonch.2017.11.029.
Kumar P S, Kumar M S, Pandit A. 2000. Experimental quantification of chemical effects of hydrodynamic cavitation. Chemical Engineering Science, 55(9):1 633-1 639, https://doi.org/10.1016/S0009-2509(99)00435-2.
Liu J G, Zhang X L, Sun Y H, Lin W. 2010.Antioxidative capacity and enzyme activity in Haematococcus pluvialis cells exposed to superoxide free radicals. Chinese Journal of Oceanology and Limnology, 28(1):1-9, https://doi.org/10.1007/s00343-010-9244-6.
Ma X L, Yu J Z, Zhu B H, Pan K H, Pan J, Yang G P. 2011.Cloning and characterization of a delta-6 desaturase encoding gene from Nannochloropsis oculata. Chinese Journal of Oceanology and Limnology, 29(2):290-296, https://doi.org/10.1007/s00343-011-0048-0.
Markou G, Nerantzis E. 2013. Microalgae for high-value compounds and biofuels production:a review with focus on cultivation under stress conditions. Biotechnology Advances, 31(8):1 532-1 542, https://doi.org/10.1016/j.biotechadv.2013.07.011.
Naveena B, Armshaw P, Pembroke J T. 2015. Ultrasonic intensification as a tool for enhanced microbial biofuel yields. Biotechnology for Biofuels, 8:140, https://doi.org/10.1186/s13068-015-0321-0.
Nugbienyo L, Malinina Y, Garmonov S, Kamencev M, Salahov I, Andruch V, Moskvin L, Bulatov A. 2017. Automated sugaring-out liquid-liquid extraction based on flow system coupled with HPLC-UV for the determination of procainamide in urine. Talanta, 167:709-713, https://doi.org/10.1016/j.talanta.2017.02.051.
Phong W N, Show P L, Teh W H, Teh T X, Lim H M Y, Nazri N S B, Tan C H, Chang J S, Ling T C. 2017. Proteins recovery from wet microalgae using liquid biphasic flotation (LBF). Bioresource Technology, 244:1 329-1 336, https://doi.org/10.1016/j.biortech.2017.05.165.
Platis D, Labrou N E. 2009. Application of a PEG/salt aqueous two-phase partition system for the recovery of monoclonal antibodies from unclarified transgenic tobacco extract.Biotechnology Journal, 4(9):1 320-1 327, https://doi.org/10.1002/biot.200800359.
Pradal D, Vauchel P, Decossin S, Dhulster P, Dimitrov K. 2016. Kinetics of ultrasound-assisted extraction of antioxidant polyphenols from food by-products:extraction and energy consumption optimization. Ultrasonics Sonochemistry, 32:137-146.
Safi C, Ursu A V, Laroche C, Zebib B, Merah O, Pontalier P Y, Vaca-Garcia C. 2014. Aqueous extraction of proteins from microalgae:effect of different cell disruption methods. Algal Research, 3:61-65, https://doi.org/10.1016/j.algal.2013.12.004.
Sankaran R, Manickam S, Yap Y J, Ling T C, Chang J S, Show P L. 2018. Extraction of proteins from microalgae using integrated method of sugaring-out assisted liquid biphasic flotation (LBF) and ultrasound. Ultrasonics Sonochemistry, 48:231-239, https://doi.org/10.1016/j.ultsonch.2018.06.002.
Selvakumar P, Ling T C, Walker S, Lyddiatt A. 2010.A practical implementation and exploitation of ATPS for intensive processing of biological feedstock:a novel approach for heavily biological feedstock loaded ATPS.
Separation and Purification Technology, 75(3):323-331, https://doi.org/10.1016/j.seppur.2010.08.022.
Shirsath S R, Sable S S, Gaikwad S G, Sonawane S H, Saini D R, Gogate P R. 2017. Intensification of extraction of curcumin from Curcuma amada using ultrasound assisted approach:effect of different operating parameters.Ultrasonics Sonochemistry, 38:437-445, https://doi.org/10.1016/j.ultsonch.2017.03.040.
Shishov A, Nechaeva D, Moskvin L, Andruch V, Bulatov A. 2017. Automated solid sample dissolution coupled with sugaring-out homogenous liquid-liquid extraction.Application for the analysis of throat lozenge samples.Journal of Molecular Liquids, 233:149-155, https://doi.org/10.1016/j.molliq.2017.03.022.
Show P L, Tan C P, Anuar M S, Ariff A, Yusof Y A, Chen S K, Ling T C. 2011. Direct recovery of lipase derived from Burkholderia cepacia in recycling aqueous two-phase flotation. Separation and Purification Technology, 80(3):577-584, https://doi.org/10.1016/j.seppur.2011.06.013.
Sicaire A G, Vian M A, Fine F, Carré P, Tostain S, Chemat F. 2016. Ultrasound induced green solvent extraction of oil from oleaginous seeds. Ultrasonics Sonochemistry, 31:319-329, https://doi.org/10.1016/j.ultsonch.2016.01.011.
Sivakumar M, Senthilkumar P, Majumdar S, Pandit A B. 2002.Ultrasound mediated alkaline hydrolysis of methyl benzoate - reinvestigation with crucial parameters.Ultrasonics Sonochemistry, 9(1):25-30, https://doi.org/10.1016/S1350-4177(01)00099-2.
Sivakumar V, Rani K, Kumari M. 2017. Efficient extraction of natural dye from red sandal wood (Pterocarpus Sandalinus) using ultrasound. International Wood Products Journal, 8(1):6-9, https://doi.org/10.1080/2042 6445.2016.1214380.
Soares R R, Azevedo A M, Van Alstine J M, Aires-Barros M R. 2015. Partitioning in aqueous two-phase systems:analysis of strengths, weaknesses, opportunities and threats.Biotechnology Journal, 10(8):1 158-1 169, https://doi.org/10.1002/biot.201400532.
Sobianowska K, Walkowiak W, Kozłowski C. 2009. Principles and applications of solvent sublation-a review. Ars Separatoria Acta, 7:23-38.
Timofeeva I, Shishov A, Kanashina D, Dzema D, Bulatov A. 2017. On-line in-syringe sugaring-out liquid-liquid extraction coupled with HPLC-MS/MS for the determination of pesticides in fruit and berry juices.Talanta, 167:761-767, https://doi.org/10.1016/j.talanta. 2017.01.008.
Tu X J, Sun F Y, Wu S Y, Liu W Y, Gao Z S, Huang S K, Chen W B. 2018. Comparison of salting-out and sugaring-out liquid-liquid extraction methods for the partition of 10-hydroxy-2-decenoic acid in royal jelly and their coextracted protein content. Journal of Chromatography B, 1073:90-95, https://doi.org/10.1016/j.jchromb.2017.12. 020.
Wang B, Ezejias T, Feng H, Blaschek H. 2008a. Sugaring-out:a novel phase separation and extraction system. Chemical Engineering Science, 63(9):2 595-2 600, https://doi.org/10.1016/j.ces.2008.02.004.
Wang B, Feng H, Ezeji T, Blaschek H. 2008b. Sugaring-out separation of acetonitrile from its aqueous solution.Chemical Engineering & Technology, 31(12):1 869-1 874, https://doi.org/10.1002/ceat.200800003.
Wu J Y, Lin L D, Chau F T. 2001. Ultrasound-assisted extraction of ginseng saponins from ginseng roots and cultured ginseng cells. Ultrasonics Sonochemistry, 8(4):347-352, https://doi.org/10.1016/S1350-4177(01)00066-9.
Xia S, Wan L L, Li A F, Sang M, Zhang C W. 2013. Effects of nutrients and light intensity on the growth and biochemical composition of a marine microalga Odontella aurita.Chinese Journal of Oceanology and Limnology, 31(6):1 163-1 173.
Yang H C, Zeng M Y, Dong S Y, Liu Z Y, Li R X. 2010.Antiproliferative activity of phlorotannin extracts from brown algae Laminaria japonica Aresch. Chinese Journal of Oceanology and Limnology, 28(1):122-130, https://doi.org/10.1007/s00343-010-9054-x.
Zhang C, Huang K, Yu P H, Liu H Z. 2012. Sugaring-out threeliquid-phase extraction and one-step separation of Pt(IV), Pd(Ⅱ) and Rh(Ⅲ). Separation and Purification Technology, 87:127-134, https://doi.org/10.1016/j.seppur.2011.11.032.
Zimmermann S, Gretzinger S, Zimmermann P K, Bogsnes A, Hansson M, Hubbuch J. 2018. Cell separation in aqueous two-phase systems- influence of polymer molecular weight and tie-line length on the resolution of five model cell lines. Biotechnology Journal, 13(2):1 700 250, https://doi.org/10.1002/biot.201700250.
Copyright © Haiyang Xuebao