สภาวะสารอาหารที่เหมาะสมสำหรับการผลิตมวลเซลล์และโอเมก้า-3 จากทรอสโทไคทริดส์
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เม.ย. 30, 2018
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ทรอสโทไคทริดส์ มวลเซลล์ กรดไขมันโอเมก้า-3
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บทคัดย่อ
งานวิจัยนี้มีวัตถุประสงค์เพื่อประเมินผลของแหล่งคาร์บอนและแหล่งไนโตรเจนต่อการผลิตมวลเซลล์และกรดไขมันโอเมก้า-3 จากทรอสโทไคทริดส์ และศึกษาสายสัมพันธ์เชิงวิวัฒนาการของเชื้อที่มีศักยภาพโดยลำดับเบสในนิวคลีโอไทด์ ศึกษาการผลิตมวลเซลล์และโอเมก้า-3 โดยอาหารที่มีกลูโคส 3 และ 6 เปอร์เซ็นต์ ที่เติมแหล่งไนโตรเจน (1 เปอร์เซ็นต์ ยีสต์สกัด, 1 เปอร์เซ็นต์ ยีสต์สกัดและเพปโตน, 1 เปอร์เซ็นต์ ยีสต์สกัด เพปโตนและทริปโตน) เพื่อประเมินผลของชุดการทดลองเหล่านี้ จึงศึกษาปริมาณมวลเซลล์และโอเมก้า-3 โดยวิเคราะห์น้ำหนักเซลล์แห้งและแก๊สโครมาโตกราฟี-แมสสเปคโตรมิทรี ตามลำดับ ผลการศึกษาพบว่าทรอสโทไคทริดส์ ไอโซเลท TC9-TSU สามารถผลิตมวลเซลล์ได้สูงสุดเท่ากับ 32.87 กรัมต่อลิตร ในอาหาร 6 เปอร์เซ็นต์ กลูโคส และ 1 เปอร์เซ็นต์ ยีสต์สกัด เพปโทน และทริปโตน นอกจากนี้เชื้อไอโซเลทนี้ยังสามารถผลิตกรดไขมันโอเมก้า-3 ได้สูงที่สุด (52.56 เปอร์เซ็นต์) โดยพบกรดไขมันดีเอชเอ (docosahexaenoic acid, 22:6n3) ดีพีเอ(docosapentaenoic acid, 22:5n3) และ อีพีเอ (eicosapentaenoic acid, 20:5n3) จากผลการจัดจำแนกทรอสโทไคทริดส์โดยการวิเคราะห์ลำดับเบสในนิวคลีโอไทด์ของยีน 18S ของไรโบโซมอลอาร์เอ็นเอ พบว่าไอโซเลท TC9-TSU จัดจำแนกได้เป็น Aurantiochytrium limacinum (รหัสนิวคลีโอไทด์ MG279107) จากผลการศึกษาแสดงให้เห็นว่าทรอสโทไคทริดส์เป็นแหล่งทางเลือกสำหรับการผลิตโอเมก้า-3 กรดไขมันไม่อิ่มตัวเชิงซ้อนที่น่าสนใจสำหรับประยุกต์ในอนาคต
คำสำคัญ : ทรอสโทไคทริดส์; มวลเซลล์; กรดไขมันโอเมก้า-3
Article Details
ประเภทบทความ
Biological Sciences
เอกสารอ้างอิง
[1] Sijtsma, L. and Swaaf, D.M., 2004, Biotechnological production and applica-tions of the omega-3 polyunsaturated fatty acid, docosahexaenoic acid, Appl. Microbiol. Biotechnol. 64: 146-153.
[2] Jain, R., Raghukumar, S., Tharanathan, R. and Bhosle, N.B., 2005, Extracellular poly-saccharide production by thraustochytrid protists, Mar. Biotechnol. 7: 184-192.
[3] Simopoulos, A.P., Kifer, R.R., Martin, R.E. and Barlaw, S.M., 1991, Health effects of omega 3 polyunsaturated fatty acids in seafoods, World Rev. Nutr. Diet. 66: 1-592.
[4] Shwu-Tzy, W., Shih-Tsung, Y. and Liang-Ping, L., 2005, Effect of culture conditions on docosahexaenoic acid production by Schizochytrium sp. S31, Proc. Biochem. 40: 3103-3108.
[5] Nauroth, J.M., Liu, Y.C. and Elswyk. M.V., 2010, Docosahexaenoic acid (DHA) and docosapentaenoic acid (DPA n-6) algal oils reduce inflammatory mediators in human peripheral mononuclear cells in vitro and paw edema in vivo, Lipids 45: 375-384.
[6] Corsinovi, L., Bias,i F., Poli, G., Leonarduzzi, G. and Isaia, G., 2011, Dietary lipids and their oxidized products in Alzheimer’s disease, Mol. Nutr. Food Res. 55: 161-172.
[7] Raghukumar, S., 2008, Thraustochytrid marine protists: Production of PUFAs and other emerging technologies, Mar. Biotechnol. 10: 631-640.
[8] Raghukumar, S. and Damare, V.S., 2011, Increasing evidence for the important role of Labyrinthulomycetes in marine ecosystems, Bot. Mar. 54: 3-11.
[9] Lewis, T.E., Nichols, P.D. and McMeekin, T.A., 1999, The biotechnological potential of thraustochytrids, Mar. Biotechnol. 1: 580-587.
[10] Leano, M.D.V., Jones, G.B.G. and Pang, K.L., 2012, Labyrinthulomycota, pp. 215-244, In Marine Fungi and Fngal-like Organisms, 1st Ed., Walter de Gruyter GmbH, Germany.
[11] Shene, C., Leyton, A., Esparza, Y., Flores, L., Quilodrán, B. and Hinzpeter, I., 2010, Microbial oils and fatty acids: Effect of carbon source on docosahexaenoic acid (c22:6 n-3, DHA) production by thraustochytrid strains, J. Soil Sci. Plant Nutr. 10: 207-216.
[12] Chamberlian, A.H.L. and Moss, S.T., 1998, The thraustochytrids: A protest groupwith mixed affinities, Biosystems 21: 341-349.
[13] Alderman, D.J., Harrison, J.L., Bremer, G.B. and Jone, E.B.G., 1974, Taxonomic revisions in the marine biflagellate fungi: The ultrastructural evidence, Mar. Biol. 25: 345-357.
[14] Porter, D., 1989. Handbook of Protoctista: Phylum Labyrinthulomycota Net Slime Mold, Boston.
[15] Honda, D., Yokochi, T., Nakahara, T., Erata, M. and Higashihara, T., 1998, Schizochytrium limacinum sp. Nov., a new thraustochytrids from mangrove area in the west Pacific Ocean, Mycol. Res. 102: 439-448.
[16] Leander, C.A., Porter, D. and Leander, B.S., 2004, Comparative morphology and molecular phylogeny of aplanochytrids (Labyrinthulomycota), Eur. J. Protistol. 40: 317-328.
[17] Yokoyama, R. and Honda, D., 2007. Taxonomic rearrangement of the genus Schizochytrium sensu lato on morphology, chemotaxonomic charac-teristics, and 18S rRNA gene phylogeny (Thraustochytriaceae, Labyrinthuromuce-tes): Emendation for Schizochytrium and erection of Aurantiochytrium and Oblongichytrium gen.nov., Mycoscience 48: 199-211.
[18] Mo, C. and Rinkevich, B., 2001, A simple, reliable, and fast protocol for thraustochytrid DNA extraction, Mar. Biotechnol. 3: 100-102.
[19] Harel, M., Ben-Dov, E., Rasoulouniriana, D., Siboni, N., Kramarsky-Winter, E., Loya, Y., Barak, Z., Wiesman, Z. and Kushmaro, A., 2008, A new thraustochytrid, strain Fng1, isolated from the surface mucus of the hermatypic coral Fungia granulosa, FEMS Microbiol. Ecol. 64: 378-387.
[20] Hall, T., 2005, BioEdit: Biological Sequence Alignment Editor for Windows 95/98/NT/ XP, Available Source: http://www.mbio.ncsu.edu/bioedit/page1.html.
[21] Edgar, R.C., 2004, MUSCLE: Multiple sequence alignment with high accuracy and high throughput, Nucl. Acids Res. 32: 1792-1797.
[22] Swofford, D.L., 2002. PAUP*: Phylogenetic Analysis Using Parsimony (*and other Methods), Version 4. Sunderland, Sinauer Associates, Massachusetts.
[23] Kishino, H. and Hasegawa, M., 1989, Evaluation of the maximum likelihood estimate of the evolutionary tree topologies from DNA sequence data, and the branching order in hominoidea, J. Mol. Evol. 29: 170-179.
[24] Nylander, J.A.A., 2004, MrModeltest v. 2.0. Evolutionary Biology Centre, Uppsala University: Program Distributed by the Author.
[25] Perveen, Z., Ando, H., Ueno, A., Ito, Y., Yamamoto, Y., Yamada, Y., Takagi, T., Kaneko, T., Kogame, K. and Okuyama, H., 2006, Isolation and characterization of a novel thraustochytrid-like microorganisms that efficiently produces docosahexae-noic acid, Biotechnol. Lett. 28: 197-202.
[26] Yang, H.L., Lu, C.K., Chen, S.F., Chen, Y.M. and Chen, Y.M., 2010, Isolation and characterisation of Taiwanese hetero-trophic microalgae: screening of strains for docosahexaenoic acid (DHA) production, Mar. Biotechnol. 12: 173-185.
[27] Bahnweg, G., 1997, Studies on the physiology of thraustochytriales I: Growth requirements and nitrogen nutrition of Thraustochytrium spp., Schizochytrium sp., Japoochytrium sp., Ulkenia spp., and Labyrinthuloides spp., Veroff. Inst. Meeresforsch. Bremerhaven 17: 245-268.
[28] Jones, E.B.G. and Harrison, J.L., 1976, Physiology of marine phycomycetes, Adv. Aqu. Mycol. 3: 261-278.
[29] Burja, A.M., Radianingtyas, H., Windust, A. and Barrow, C.J., 2006, Isolation and characterization of polyunsaturated fatty acid producing Thraustochytrium species: Screening of strains and optimization of omega-3 production, Appl. Microbiol. Biotechnol. 72: 1161-1169.
[30] Ganuza, E. and Izquierdo, M., 2007, Lipid accumulation in Schizochytrium G13/2S production in continuous culture, Appl. Microbiol. Biotechnol. 76: 985-990.
[31] Wong, M.K.M., Tsui, C.K.M., Au, D.W.T. and Wrijmoed, L.L.P., 2008, Docosahexaenoic acid production and ultrastructure of the thraustochytrid Aurantiochytrium mangrovei MP2 under high glucose concentration, Mycoscience. 49: 266-270.
[32] Ren, L.J., Huang, H., Xiao, A.H., Lian, M., Jin, L.J. and Ji, X.J. 2009, Enhanced docosahexaenoic acid production by reinforcing acetyl-CoA and NADPH supply in Schizochytrium sp. HX- 308, Bioprocess Biosyst. Eng. 32: 837-843.
[33] Zhou, P.P., Lu, M.B., Li, W. and Yu, L.J., 2010, Microbial production of docosahexaenoic acid by a low temperature-adaptive strain Thraustochytriidae sp. Z105: Screening and optimization, J. Basic Microbiol. 50: 380-387.
[34] Xiong, Z.G., 1995, Fermentation: Process and Theory, China Medical Science and Technology Press.
[35] Wen, Z.Y. and Chen, F., 2001, Optimiza-tion of nitrogen sources for heterotrophic production of eicosapentaenoic acid by the diatom Nitzschia laevis, Enzyme Microb. Technol. 29, 341-347
[36] Fan, K.W. and Chen, F., 2007, Production of High-Value Products by Marine Microalgae Thraustochytrids, pp. 293- 324, In Yang, S.T. (Ed.), Bioprocessing for Value-Added Products from Renewable Resources: New Technologies and Applications, Elsevier.
[37] Chen, G.Q., Fan, K.W., Lu, F.P., 2010, Optimization of nitrogen source for enhanced production of squalene from Thraustochytrid Aurantiochytrium sp., New Biotechnol. 27: 382-389.
[38] Shene, C., Leyton, A. Esparza, Y., Flores, L., Quilodram, B., Hinzpeter, I. and Rubilar, M., 2010, Microbial oils and fatty acids: effect of carbon source on docosahexaenoic acid (C22:6 N-3, DHA) production by thraustochytrid strain, J. Soil Sci. Plant Nutr. 10: 207-216.
[39] Nakazawa, A., Matsuura, H., Kose, R., Ito, K., Uedo, M., Honda, D., Inouye, I., Kaya K. and Watanabe, M.M., 2012, Optimization of biomass and fatty acid production by Aurantiochytrium sp. Strain 4W-1b, Proc. Environ. Sci. 15: 27-33.
[40] Fan, K.W., Vrijmoed, L.L.P. and Jone, E.B.G., 2002, Physiological studies of subtropical mangrovei thraustochytrids, Bot. Mar. 45: 50-57.
[41] Fan, K.W. and Kamlangdee, N., 2003, Polyunsaturated fatty acids production by Schizochytrium sp. Isolated from mangrove, J. Sci. Technol. 25: 643-650.
[42] Chang, K.J.L., Nichols, C.M., Blackburn, S.I., Dunstan, G.A., Koutoulis, A. and Nichols, P.D., 2014, Comparison of Thraustochy-trium sp., Schizochytrium sp., Thraustochytrium sp. and Ulkenia sp. for production of biodiesel, long-chain omega-3 oils, and exopolysaccharide, Mar. Biotechnol. 16: 396-411.
[43] Manikan, V., Kalil, M.S. and Hamid, A.A., 2015, Response surface optimization of culture medium for enhanced docosahexaenoic acid production by a Malaysian thraustochytrid, Sci. Rep. 5: 1-8.
[44] Furlan, V.J.M., Maus, V., Batista, I. and Bandarra, N.M., 2017, Production of docosahexaenoic acid by Aurantiochy-trium sp. ATCC PRA-276, Braz. J. Microbiol. 48: 359-365.
[45] Homayooni, B., Sahari, M.A. and Barzegar, M., 2014, Concentration of omega-3 fatty acids from rainbow sardine fish oil by various methods, IFRJ. 21: 743-748.
[2] Jain, R., Raghukumar, S., Tharanathan, R. and Bhosle, N.B., 2005, Extracellular poly-saccharide production by thraustochytrid protists, Mar. Biotechnol. 7: 184-192.
[3] Simopoulos, A.P., Kifer, R.R., Martin, R.E. and Barlaw, S.M., 1991, Health effects of omega 3 polyunsaturated fatty acids in seafoods, World Rev. Nutr. Diet. 66: 1-592.
[4] Shwu-Tzy, W., Shih-Tsung, Y. and Liang-Ping, L., 2005, Effect of culture conditions on docosahexaenoic acid production by Schizochytrium sp. S31, Proc. Biochem. 40: 3103-3108.
[5] Nauroth, J.M., Liu, Y.C. and Elswyk. M.V., 2010, Docosahexaenoic acid (DHA) and docosapentaenoic acid (DPA n-6) algal oils reduce inflammatory mediators in human peripheral mononuclear cells in vitro and paw edema in vivo, Lipids 45: 375-384.
[6] Corsinovi, L., Bias,i F., Poli, G., Leonarduzzi, G. and Isaia, G., 2011, Dietary lipids and their oxidized products in Alzheimer’s disease, Mol. Nutr. Food Res. 55: 161-172.
[7] Raghukumar, S., 2008, Thraustochytrid marine protists: Production of PUFAs and other emerging technologies, Mar. Biotechnol. 10: 631-640.
[8] Raghukumar, S. and Damare, V.S., 2011, Increasing evidence for the important role of Labyrinthulomycetes in marine ecosystems, Bot. Mar. 54: 3-11.
[9] Lewis, T.E., Nichols, P.D. and McMeekin, T.A., 1999, The biotechnological potential of thraustochytrids, Mar. Biotechnol. 1: 580-587.
[10] Leano, M.D.V., Jones, G.B.G. and Pang, K.L., 2012, Labyrinthulomycota, pp. 215-244, In Marine Fungi and Fngal-like Organisms, 1st Ed., Walter de Gruyter GmbH, Germany.
[11] Shene, C., Leyton, A., Esparza, Y., Flores, L., Quilodrán, B. and Hinzpeter, I., 2010, Microbial oils and fatty acids: Effect of carbon source on docosahexaenoic acid (c22:6 n-3, DHA) production by thraustochytrid strains, J. Soil Sci. Plant Nutr. 10: 207-216.
[12] Chamberlian, A.H.L. and Moss, S.T., 1998, The thraustochytrids: A protest groupwith mixed affinities, Biosystems 21: 341-349.
[13] Alderman, D.J., Harrison, J.L., Bremer, G.B. and Jone, E.B.G., 1974, Taxonomic revisions in the marine biflagellate fungi: The ultrastructural evidence, Mar. Biol. 25: 345-357.
[14] Porter, D., 1989. Handbook of Protoctista: Phylum Labyrinthulomycota Net Slime Mold, Boston.
[15] Honda, D., Yokochi, T., Nakahara, T., Erata, M. and Higashihara, T., 1998, Schizochytrium limacinum sp. Nov., a new thraustochytrids from mangrove area in the west Pacific Ocean, Mycol. Res. 102: 439-448.
[16] Leander, C.A., Porter, D. and Leander, B.S., 2004, Comparative morphology and molecular phylogeny of aplanochytrids (Labyrinthulomycota), Eur. J. Protistol. 40: 317-328.
[17] Yokoyama, R. and Honda, D., 2007. Taxonomic rearrangement of the genus Schizochytrium sensu lato on morphology, chemotaxonomic charac-teristics, and 18S rRNA gene phylogeny (Thraustochytriaceae, Labyrinthuromuce-tes): Emendation for Schizochytrium and erection of Aurantiochytrium and Oblongichytrium gen.nov., Mycoscience 48: 199-211.
[18] Mo, C. and Rinkevich, B., 2001, A simple, reliable, and fast protocol for thraustochytrid DNA extraction, Mar. Biotechnol. 3: 100-102.
[19] Harel, M., Ben-Dov, E., Rasoulouniriana, D., Siboni, N., Kramarsky-Winter, E., Loya, Y., Barak, Z., Wiesman, Z. and Kushmaro, A., 2008, A new thraustochytrid, strain Fng1, isolated from the surface mucus of the hermatypic coral Fungia granulosa, FEMS Microbiol. Ecol. 64: 378-387.
[20] Hall, T., 2005, BioEdit: Biological Sequence Alignment Editor for Windows 95/98/NT/ XP, Available Source: http://www.mbio.ncsu.edu/bioedit/page1.html.
[21] Edgar, R.C., 2004, MUSCLE: Multiple sequence alignment with high accuracy and high throughput, Nucl. Acids Res. 32: 1792-1797.
[22] Swofford, D.L., 2002. PAUP*: Phylogenetic Analysis Using Parsimony (*and other Methods), Version 4. Sunderland, Sinauer Associates, Massachusetts.
[23] Kishino, H. and Hasegawa, M., 1989, Evaluation of the maximum likelihood estimate of the evolutionary tree topologies from DNA sequence data, and the branching order in hominoidea, J. Mol. Evol. 29: 170-179.
[24] Nylander, J.A.A., 2004, MrModeltest v. 2.0. Evolutionary Biology Centre, Uppsala University: Program Distributed by the Author.
[25] Perveen, Z., Ando, H., Ueno, A., Ito, Y., Yamamoto, Y., Yamada, Y., Takagi, T., Kaneko, T., Kogame, K. and Okuyama, H., 2006, Isolation and characterization of a novel thraustochytrid-like microorganisms that efficiently produces docosahexae-noic acid, Biotechnol. Lett. 28: 197-202.
[26] Yang, H.L., Lu, C.K., Chen, S.F., Chen, Y.M. and Chen, Y.M., 2010, Isolation and characterisation of Taiwanese hetero-trophic microalgae: screening of strains for docosahexaenoic acid (DHA) production, Mar. Biotechnol. 12: 173-185.
[27] Bahnweg, G., 1997, Studies on the physiology of thraustochytriales I: Growth requirements and nitrogen nutrition of Thraustochytrium spp., Schizochytrium sp., Japoochytrium sp., Ulkenia spp., and Labyrinthuloides spp., Veroff. Inst. Meeresforsch. Bremerhaven 17: 245-268.
[28] Jones, E.B.G. and Harrison, J.L., 1976, Physiology of marine phycomycetes, Adv. Aqu. Mycol. 3: 261-278.
[29] Burja, A.M., Radianingtyas, H., Windust, A. and Barrow, C.J., 2006, Isolation and characterization of polyunsaturated fatty acid producing Thraustochytrium species: Screening of strains and optimization of omega-3 production, Appl. Microbiol. Biotechnol. 72: 1161-1169.
[30] Ganuza, E. and Izquierdo, M., 2007, Lipid accumulation in Schizochytrium G13/2S production in continuous culture, Appl. Microbiol. Biotechnol. 76: 985-990.
[31] Wong, M.K.M., Tsui, C.K.M., Au, D.W.T. and Wrijmoed, L.L.P., 2008, Docosahexaenoic acid production and ultrastructure of the thraustochytrid Aurantiochytrium mangrovei MP2 under high glucose concentration, Mycoscience. 49: 266-270.
[32] Ren, L.J., Huang, H., Xiao, A.H., Lian, M., Jin, L.J. and Ji, X.J. 2009, Enhanced docosahexaenoic acid production by reinforcing acetyl-CoA and NADPH supply in Schizochytrium sp. HX- 308, Bioprocess Biosyst. Eng. 32: 837-843.
[33] Zhou, P.P., Lu, M.B., Li, W. and Yu, L.J., 2010, Microbial production of docosahexaenoic acid by a low temperature-adaptive strain Thraustochytriidae sp. Z105: Screening and optimization, J. Basic Microbiol. 50: 380-387.
[34] Xiong, Z.G., 1995, Fermentation: Process and Theory, China Medical Science and Technology Press.
[35] Wen, Z.Y. and Chen, F., 2001, Optimiza-tion of nitrogen sources for heterotrophic production of eicosapentaenoic acid by the diatom Nitzschia laevis, Enzyme Microb. Technol. 29, 341-347
[36] Fan, K.W. and Chen, F., 2007, Production of High-Value Products by Marine Microalgae Thraustochytrids, pp. 293- 324, In Yang, S.T. (Ed.), Bioprocessing for Value-Added Products from Renewable Resources: New Technologies and Applications, Elsevier.
[37] Chen, G.Q., Fan, K.W., Lu, F.P., 2010, Optimization of nitrogen source for enhanced production of squalene from Thraustochytrid Aurantiochytrium sp., New Biotechnol. 27: 382-389.
[38] Shene, C., Leyton, A. Esparza, Y., Flores, L., Quilodram, B., Hinzpeter, I. and Rubilar, M., 2010, Microbial oils and fatty acids: effect of carbon source on docosahexaenoic acid (C22:6 N-3, DHA) production by thraustochytrid strain, J. Soil Sci. Plant Nutr. 10: 207-216.
[39] Nakazawa, A., Matsuura, H., Kose, R., Ito, K., Uedo, M., Honda, D., Inouye, I., Kaya K. and Watanabe, M.M., 2012, Optimization of biomass and fatty acid production by Aurantiochytrium sp. Strain 4W-1b, Proc. Environ. Sci. 15: 27-33.
[40] Fan, K.W., Vrijmoed, L.L.P. and Jone, E.B.G., 2002, Physiological studies of subtropical mangrovei thraustochytrids, Bot. Mar. 45: 50-57.
[41] Fan, K.W. and Kamlangdee, N., 2003, Polyunsaturated fatty acids production by Schizochytrium sp. Isolated from mangrove, J. Sci. Technol. 25: 643-650.
[42] Chang, K.J.L., Nichols, C.M., Blackburn, S.I., Dunstan, G.A., Koutoulis, A. and Nichols, P.D., 2014, Comparison of Thraustochy-trium sp., Schizochytrium sp., Thraustochytrium sp. and Ulkenia sp. for production of biodiesel, long-chain omega-3 oils, and exopolysaccharide, Mar. Biotechnol. 16: 396-411.
[43] Manikan, V., Kalil, M.S. and Hamid, A.A., 2015, Response surface optimization of culture medium for enhanced docosahexaenoic acid production by a Malaysian thraustochytrid, Sci. Rep. 5: 1-8.
[44] Furlan, V.J.M., Maus, V., Batista, I. and Bandarra, N.M., 2017, Production of docosahexaenoic acid by Aurantiochy-trium sp. ATCC PRA-276, Braz. J. Microbiol. 48: 359-365.
[45] Homayooni, B., Sahari, M.A. and Barzegar, M., 2014, Concentration of omega-3 fatty acids from rainbow sardine fish oil by various methods, IFRJ. 21: 743-748.
