Growth and Fatty Acid Profile of Thraustochytrium sp.
CR01 Using Different Sugar Substitutes
Marigold O. Uba1, Katherine Charmaine P. Duabe2,3,
Maria Amabelle Christine M. Biene2,3, Ma. Kristina Celyna R. Ortiz2,3,
Reuel M. Bennett2, and Gina R. Dedeles1,2,3*
1The Graduate School, 2Research Center for the Natural and Applied Sciences,
Thomas Aquinas Research Complex and 3Department of Biological Sciences,
College of Science, University of Santo Tomas, España, Manila 1015 Philippines
*corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.
ABSTRACT
Thraustochytrids, which are a group of marine heterokonts, have shown their promising potential as a good source of the omega-3 polyunsaturated fatty acids (PUFAs) docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) of importance to human health and aquaculture, respectively. A cost-effective production method (using alternative carbon source) of these thraustochytrids coupled with higher PUFA yield has yet to be established for commercial exploitation of lipids. In this study, Thraustochytrium sp. CR01 isolated from senescent fallen mangrove leaves in Coastal Road, Cavite was grown on different sugar products such as liquid sugar, molasses, and corn syrup as glucose substitute for carbon source in the culture medium. Based on growth analysis, there is no significant difference between the sugar used in which the corn syrup biomass produced 0.15 – 0.25 g/ 25mL; while liquid sugar and molasses had 0.1 – 0.3 g/ 25mL. Analysis of fatty acid methyl ester results showed that Thraustochytrium sp. CR01 produced predominantly palmitic acid (16:0), a saturated fatty acid which constitutes 57% total fatty acids (TFA) in corn syrup, 54% TFA in molasses, and 25% TFA in liquid sugar. DHA was also produced at 5% TFA in corn syrup, 7% TFA in molasses, and 2% TFA in liquid sugar.
INTRODUCTION
Thraustochytrids are a large group of marine heterokonts which belong to Kingdom Straminipila and Phylum Labyrinthulomycetes (Arafiles et al. 2011). Although ubiquitous, studies on these group of organisms are still limited. Biomass of these ubiquitous microorganisms is 10-50% total oil and 30-70% of which is DHA (Singh & Ward 2005). Moreover, thraustochytrids are capable of producing up to 90% monounsaturated fatty acids (MUFAs) such as palmitoleic acid (C16:1), oleic acid (C18:1), eicosenoic acid (C20:1), and erucic acid (C22:1). This gives the thraustochytrids another feature of world-wide significance – as a source of fatty acids for biodiesel production (Fisher et al. 2008). . . . . . read more
REFERENCES
Aki T, Hachida K, Yoshinaga M, Katai Y, Yamasaki T, Kawamoto S, Kakizono T, Maoka T, Shigeta S, Suzuki O, Ono K. 2003. Thraustochytrid as a potential source of carotenoids. Journal of the American Oil Chemist’s Society 80:789-794.
Aki T, Iwasaka H, Adachi H, Watanabe K, Kawamoto S, Ono K. 2013. Utilization of waste syrup for production of polyunsaturated fatty acids and xanthophylls by Aurantiochytrium. Journal of Oleo Science 62 (9):729-736.
Arafiles KHV, Alcantara JCO, Cordero PRF, Batoon JAL, Galura, FS, Leaño EM, Dedeles GR. 2011. Cultural optimization of thraustochytrids for biomass and fatty acid production. Mycosphere 2(5):521-531.
Ashford A, Barclay WR, Weaver CA, Giddings TH, Zeller S. 2000. Electron microscopy may reveal structure of docosahexaenoic acid-rich oil within Schizochytrium sp. Lipids 35:1377–1386.
BAJPAI PK, BAJPAI P, WARD OP. 1991. Optimization of production of docosahexaenoic acid (DHA) by Thraustochytrium aureum ATCC 34304. Journal of American Oil Chemist’s Society 68:509–514.
BONGIORNI L, JAIN R, RAGHUKUMAR S, AGGARWAL RK. 2005. Thraustochytrium gaertnerium sp. nov.: a new thraustochytrid stramenopilan protist from mangroves of Goa, India. Protist 156:303–315.
BOWLES RD, HUNT AE, BREMER GB, DUCHARS MG, EATON RA. 1999. Long-chain n-3 polyunsaturated fatty acid production by members of the marine protistan group, the thraustochytrids: screening of isolates and optimisation of docosahexaenoic acid production. Journal of Biotechnology 70: 193-202.
CHATDUMRONG W, YONGMANITCHAI W, LIMTONG S, WORAWATTANAMATEEKUL W. 2007. Optimization of docosahexaenoic acid (DHA) production and improvement of astaxanthin content in a mutant Schizochytrium limacium isolated from mangrove forest in Thailand. Kasetsart Journal 41:324-334.
CHIHIB NE, TIERNY Y, MARY P, HORNEZ JP. 2005. Adaptational changes in cellular fatty acid branching and unsaturation of Aeromonas species as a response to growth temperature and salinity. International Journal of Food Microbiology 102:113–119.
Fan KW, Chen F, Jones EBG, Vrijmoed LLP. 2000. Utilization of food processing waste by Thraustochytrids. Fungal Diversity 5:185-194.
Fan KW, Chen F, Jones EBG, Vrijmoed LLP. 2001. Eicosapentaenoic and docosahexaenoic acids production by and okara-utilizing potential of thraustochytrids. Journal of Industrial Microbiology and Biotechnology 27:199-202.
Fan KW, Vrijmoed LLJ, Jones EBG. 2002. Physiological studies of subtropical mangrove thraustochytrids. Botanica Marina 45:50-57.
FAN KW, KAMLANGDEE N. 2003. Polyunsaturated fatty acids production by Schizochytrium sp. isolated from mangrove. Journal of Science and Technology 25:643–650.
FAN KW, CHEN F. 2007. Production of high-value products by marine microalgae thraustochytrids. In: Yang ST, editor. Bioprocessing for value added products from renewable resources. Amsterdam: Elsevier. p 293–324.
Fan K W, Jiang Y, Ho L, Chen F. 2009. Differentiation in fatty acid profiles of pigmented and nonpigmented Aurantiochytrium isolated from Hongkong mangroves. Journal of Agricultural and Food Chemistry 30(20):768-775.
Fisher L, Nicholls D, Sanderson K. 2008. Production of biodiesel. World Intellectual Property Organization 067605.
FUKUDA H, KONDO A, NODA H. 2001. Biodiesel fuel production by transesterification of oils. Journal of Bioscience and Bioengineering 92:405-416.
Genuis SJ. 2008. To sea or not to sea: Benefits and risks of gestational fish consumption. Reproductive Toxicology 26(2):81-85.
JOLLEY A. 2006. The supply of fossil fuels. Climate Change Project Working Paper Series. Victoria University of Technology: US.
Leaño EM. 2001. Straminipilous organisms from fallen mangrove leaves from Panay Island, Philippines. Fungal Diversity 6:75-81.
LEAÑO EM, GAPASIN RSJ, POLOHAN B, VRIJMOED LLP. 2003. Growth and fatty acid production of thraustochytrids from Panay mangroves, Philippines. Fungal Diversity 12:111-122.
Leaño EM, Gapasin RS, Estudillo-del Castillo C. 2009. Enrichment potential of HUFA-rich thraustochytrids Schizochytrium mangrovei for the rotifer Branchionus plicatilis. Aquaculture 293:57-61.
LEWIS TE, NICHOLS PD, MCMEEKIN TA. 1999. The Biotechnological Potential of Thraustochytrids. Marine Biotechnology 1:580-587.
LEWIS TE, NICHOLS PD, MCMEEKIN TA. 2001. Sterol and squalene content of a docosahexaenoic acid producing thraustochytrids: influence of culture, age temperature and dissolved oxygen. Marine Biotechnology 3:439–447.
LI ZY, WARD OP. 1994. Production of docosahexaenoic acid (DHA) by Thraustochytrium roseum. Journal of Industrial Microbiology 13:238–341.
PERVEEN Z, ANDO H, UENO A, ITO Y, YAMAMOTO Y, YAMADA Y, TAKAGI T, KANEKO T, KOGAME K, OKUYAMA H. 2006. Isolation and characterization of a novel thraustochytrid-like microorganisms that efficiently produces decosahexaenoic acid. Biotechnology Letters 28:1997-202.
QUILODRÁN B, HINZPETER I, QUIROZ A, SHENE C. 2009. Evaluation of liquid residues from beer and potato processing for the production of docosahexanoic acid (C20:6n-3, DHA) by native thraustochytrid strains. World Journal of Microbiology and Biotechnology 25:2121-2128.
QUILODRÁN B, HINZPETER I, HORMAZABAL E, QUIROZ A, SHENE C. 2010. Docosahexaenoic acid (C22:6n−3, DHA) and astaxanthin production by Thraustochytriidae sp. AS4-A1 a native strain with high similitude to Ulkenia sp.: Evaluation of liquid residues from food industry as nutrient sources. Enzyme and Microbial Technology 47:24-30.
RAGHUKUMAR S. 2002 Ecology of the marine protists, the Labyrinthulomycetes (Thraustochytrids and Labyrinthulids). European Journal of Protistology 38:127–145.
RAGHUKUMAR S. 2008. Thraustochytrid marine protists:Production of PUFAs and other emerging technologies. Marine Biotechnology 10(6):631-640.
SHENE C, LEYTON A, ESPARZA Y, FLORES L, QUILODRÁN B, HINZPETER I. RUBILAR M. 2010. Microbial oils and fatty acids: effect of carbon source on docosahexaenoic acid (C22:6 N-3, DHA) production by thraustochytrids strains. Journal of Soil Science and Plant Nutrition 10 (3):207 - 216.
SIJTSMA L, DE SWAAF ME. 2004. Biotechnological production and applications of the ω-3 polyunsaturated fatty acid docosahexaenoic acid. Applied Microbiology and Biotechnology 64:146-153.
SINGH A, WILSON S, WARD OP. 1996. Docosahexaenoic acid (DHA) production by Thraustochytrium sp. ATCC 20892. Journal of Microbiology and Biotechnology 12:76–83.
SINGH A, WARD OP. 2005. Omega-3/6 fatty acids: Alternative sources of production. Process Biochemistry 40: 3627-3652.
TAOKA Y, NAGANO N, OKITA Y, IZUMIDA H, SUGIMOTO S, HAYASHI M. 2009. Influences of culture temperature on the growth, lipid content and fatty acid composition of Aurantiochytrium sp. strain mh0186. Marine Biotechnology 11:368-374.
UNAGULP,ASSANTACHAIC, PHADUNGRUENGLUIJI S, SUPHANTHARIKA M, TANTICHAROEN M, VERDUYN C. 2007. Coconut water as a medium additive for the production of docosahexanoic acid (C22:6 n3) by Schizochytrium mangrovei Sk-02. Bioresource Technology 98:281-287.
WADA M, FUKUNAGA N, SASAKI S. 1987. Effect of growth temperature on phospholipid and fatty acid composition in a phychrotrophic bacterium. Plant and Cell Physiology 28:1209–1217.
WU S, YU S, LIN L. 2005. Effect of culture conditions on docosahexaenoic acid production by Schizochytium sp. S31. Process Biochemistry 40 (9): 3103-3108.
YAGUCHI T, TANAKA S, YOKOCHI T, NAKAHARA T, HIGASHIHARA T. 1997. Production of high yields of docosahexaenoic acid by Schizochytrium sp. strain SR21. Journal of American Oil Chemist’s Society 74:1431–1434.
YAMASAKI T, AKI T, SHINOZAKI M, TAGUCHI M, KAWAMOTO S, ONO K. 2006. Utilization of Shochu distillery wastewater for production of polyunsaturated fatty acids and xanthophylls using thraustochytrids. Journal of Bioscience and Bioengineering 102 (4): 323-327.
YOKOCHI T, HONDA D, HIGASHIHARA T, NAKAHARA T. 1998. Optimization of docosahexaenoic acid production by Schizochytrium limacinum SR21. Applied Microbiology and Biotechnology 49:72–76.
