สภาวะสารอาหารที่เหมาะสมสำหรับการผลิตมวลเซลล์และโอเมก้า-3 จากทรอสโทไคทริดส์

Main Article Content

พฤทฐิภร ศุภพล
ชุติมา แก้วพิบูลย์
สิตา ปรีดานนท์

Abstract

This research aimed to evaluate the effect of carbon and nitrogen sources on the production of cell mass and omega-3 fatty acids by thraustochytrids and phylogenetic relationships of the potential isolates based on their nucleotide sequences. Thraustochytrids were studied for biomass and omega-3 fatty acids production by using 3 and 6 % glucose concentrations supplemented with nitrogen from different sources, including 1 % yeast extract, 1 % yeast extract and peptone, and 1 % yeast extract, peptone and tryptone. To assess the effects of these different treatments, we determined the cell mass and omega-3 fatty acids concentration by dried cells weight and gas chromatography–mass spectrometry, respectively. The maximum dried cell weight (32.87 g/L) was obtained from isolate TC9-TSU using initial concentrations of 6 % glucose and 1 % yeast extract, peptone and tryptone. Under these conditions, this isolate gave the highest level of PUFAs production (52.56 %), with DHA (docosahexaenoic acid, 22:6n3), DPA (docosapentaenoic acid, 22:5n3), and EPA (eicosapentaenoic acid, 20:5n3). In addition, the potential isolate TC9-TSU was identified based on 18S rDNA nucleotide sequences as Aurantiochytrium limacinum (accession number MG279107). This result showed that thraustochytrids are an interested alternative source of omega-3 polyunsaturated fatty acids production for further application. 


Keywords: thraustochytrids; cell mass; omega-3 fatty acid

Downloads

Download data is not yet available.

Article Details

Section
วิทยาศาสตร์ชีวภาพ
Author Biographies

พฤทฐิภร ศุภพล

สาขาวิชาชีววิทยา คณะวิทยาศาสตร์ มหาวิทยาลัยทักษิณ วิทยาเขตพัทลุง ตำบลบ้านพร้าว อำเภอป่าพะยอม จังหวัดพัทลุง 93110

ชุติมา แก้วพิบูลย์

สาขาวิชาชีววิทยา คณะวิทยาศาสตร์ มหาวิทยาลัยทักษิณ วิทยาเขตพัทลุง ตำบลบ้านพร้าว อำเภอป่าพะยอม จังหวัดพัทลุง 93110

สิตา ปรีดานนท์

ศูนย์พันธุวิศวกรรมและเทคโนโลยีชีวภาพแห่งชาติ (ไบโอเทค) ตำบลคลองหนึ่ง อำเภอคลองหลวง จังหวัดปทุมธานี 12120

References

[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.