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Schizochytrium Sp.) AS a SOURCE of DHA

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Schizochytrium Sp.) AS a SOURCE of DHA The Potential of Heterotrophic Microalgae.............. (A.R. Hakim) THE POTENTIAL OF HETEROTROPHIC MICROALGAE (Schizochytrium sp.) AS A SOURCE OF DHA Potensi Mikroalga Heterotroph (Schizochytrium sp.) sebagai Sumber DHA Arif Rahman Hakim 1) Research Institute for Fisheries Post-harvest Mechanization; *Correspondence author: Arif Rahman Hakim, KS. Tubun Petamburan VI Jakarta Pusat 10260, E-mail: [email protected] ABSTRACT Docosahexanoic acid (DHA) is commercially obtained from marine fish. With an increasing human population, the supplies of DHA are still not sufficient to meet the world’s need of DHA as food supplement. The objective of this review is to discuss Schizochytrium sp., one of microalgae which is rich in DHA, as one of the best candidate as producer of sustainable and affordable DHA. Heterotrophic microalgae, especially genus Schizochytrium, produces omega-3 fatty acids up to 40% of total unsaturated fatty acids. Cultivation of the microalgae is easy as it does not require sunlight as source of energy. Previous publication reported that several local strains of Schizochytrium have been isolated from mangrove area in Indonesia. We expect that those strains can be cultivated in mass production as producer of DHA. Keywords: docosahexanoic acids, fatty acids, Schizochytrium sp. ABSTRAK Docosahexanoic acid (DHA) komersial sebagian besar diperoleh dari ikan laut. Dengan meningkatnya populasi penduduk dunia, sumber ini tidak akan mencukupi permintaan dari masyarakat terhadap DHA sebagai salah satu makanan kesehatan. Tujuan dari review ini adalah untuk mendiskusikan Schizochytrium sebagai salah satu mikroalga yang kaya DHA, sebagai kandidat mikroalga yang mampu memproduksi DHA secara berkelanjutan dan terjangkau. Jenis mikroalga yang bersifat heterotrof terutama genus Schizochytrium mampu menghasilkan asam lemak omega 3 mencapai 40% dari total asam lemak tak jenuhnya. Kultivasi mikroalga ini juga lebih mudah karena tidak membutuhkan sinar matahari sebagai sumber energinya. Publikasi sebelumnya menunjukkan bahwa beberapa strain lokal telah di isolasi dari daerah mangrove di Indonesia. Kami berharap strain tersebut bisa dikembangkan ke produksi DHA secara masal. Kata Kunci: asam docosaheksanoat, asam lemak, Schizochytrium sp. INTRODUCTION taste, and poor oxidative stability (Spolaore et al., 2006). As consequence, various studies should be Docosahexanoic acid (DHA; 22:6(n-3)) and conducted continuously in order to find better quality Eicosapentanoic acid (EPA ; 20:5(n-3)) are long chain and sustainable sources of omega-3 (De Swaaf et polyunsaturated fatty acids that is referred to omega al., 2003). 3. Both of these fatty acids have beneficial effect to Some marine microalgae such as dinoflagellates the human body such as maintaining cardiovascular and species in the Heterokonta phylum contain high health, preventing or treating arteriosclerosis and DHA (Barclay et al., 1994; Apt & Behrens, 1999; De certain cancer (Nettleton, 1995; Ward & Singh, 2005). Swaaf et al., 2003; Wu & Lin, 2003; Wu et al., 2005). Marine fish such as salmon, mackerel, and tuna However, majority of those microalgae are are sources of omega-3. On the other hand, over photoautotrophic that dependent on light as energy fishing, pollution, and global climate change caused source and on weather conditions. Heterotrophic scarcity in the sustainability of omega 3 sources microalgae are able to take energy from simple organic (Lewis et al., 1999). Moreover, the application of fish substances without requiring light (Apt & Behrens, oil as food additive is limited due to problems 1999). One of heterotrophic microalgae is associated with its smell characteristic, unpleasant Schizochytrium sp. which can be utilized as an 29 Squalen Vol. 7 No. 1, May 2012: 29-38. alternative to replace of fish oils due to its rapid growth to the human body, especially as the prevention of rate; its weather condition independent and its DHA several diseases such as cancer (Carrol, 1991; content which reach up to 48.95% of its total fat (Ren Zerouga et al.,1996). DHA is compiler of phospholipids et al., 2010). This paper will discuss the potential of membrane in the human brain and retina. Therefore, heterotrophic microalgae (Schizochytrium sp.) as a pregnant women and infants need to have sufficient source of DHA fatty acids. DHA intake, especially when pregnancy reaches the Recently, one of food supplement industry, Martek, age of third trimester in which the average stage produced omega-3 oil from a different strain of development of the brain and retina is the fastest. Infant Schizochytrium sp., named Algal Oil which containing could fulfill DHA diet by breast milk; however the breast approximately 37% of DHA and 16% of EPA (w/w). milk is influenced by the mother’s diet (Brenna et al., Algal Oil, as a source of DHA and EPA is intended for 2007). According to Hoffman et al. (2009) fortification food ingredient use and dietary supplement (Fedorova of milk with DHA 0.15-0.36% from the total fatty acids et al., 2011a). A number of studies including sub in the milk improves nutritional quality of infant milk. chronic feeding to rodents (Hammond et al., 2001) and non-rodents (Abril et al., 2003), developmental TAXONOMY AND MORPHOLOGY OF toxicity in rodent and non rodent species (Hammond Schizochytrium et al., 2001b), reproduction (Hammond et al., 2001c), and in vitro mutagenicity and genotoxicity (Hammond Schizochytrium is a heterotrophic microalgae et al., 2002) confirmed the safety of DHA-rich dried belongs to the family of Thraustochytriacaea (Luying Schizochytrium sp. Recently, DHA algal oil derived et al., 2008). Schizochytrium is a spherical unicellular from Schizochytrium sp. is generally recognize as microorganism. This microorganism has been favored safe (GRAS) and available for food use and for dietary for study as it is much easier to cultivate than other supplements (FDA, 2004). (Wu et al., 2005). According to Yokoyama et al. (2007), morphological characteristics of Schizochytrium under DHA AND BENEFITS the microscope would show a ectoplasmic nets, a formation of zoospores, aplanospores, and amoeboid DHA (Docosahexaenoic acids) is a type of cells, and the size between 10-20 μm. The taxonomy polyunsaturated fatty acids that has chemical detail of Schizochytrium is as follows: structure of C22: 6n-3, which is also called as derived omega 3 fatty acids. This fatty acids become important Kingdom : Chromista (Stramenopilia) since it was part of essential fatty acids that humans must ingeste because the body requires them for good Phylum : Heterokonta health but cannot synthesize them. One rich source Class : Thraustochytridae of DHA is fish oil. The high amount of DHA in fish oil is originated from photosynthetic and heterotrophic Order : Thraustochytriales microalgae consumed by the fish (Anon., 2009). Family : Thraustochytriaceae Currently, DHA is also commercially manufactured from microalgae i.e. Crypthecodinium cohnii and other Genus : Schizochytrium microalgae from the genus of Schizochytrium. Species : Schizochytrium sp. As shown in Fig. 1, the chemical structure of DHA (Source: Leipe et al., 1994) consist of 22 carbon atoms and 6 double bonds is the longest and the most unsaturated, and therefore Schizochytrium produces biflagellate zoospore and perhaps the most influential. Among the member of the mature cells divided by repeated binary division the omega-3 group of polyunsaturated fatty acids to form diads, tetrads and clusters (Fig. 2). Each (PUFA), DHA is the longest and the most unsaturated, Schizochytrium cell could develop into a sporangium therefore perhaps DHA is the most influential PUFA that produces several zoospores (Kamlangdee & Fan, (Stillwell et al., 2005). DHA has a tremendous benefit 2003). Figure 1. Structure of Docosahexaenoic acid /DHA, 22:6Ä4, 7, 10, 13, 16, 19 (Source: Stillwell et al., 2005). 30 The Potential of Heterotrophic Microalgae.............. (A.R. Hakim) Figure 2. Morfology of Schizochytrium cell (Source: Takao et al., 2005). ISOLATION OF Schizochytrium incubated for 4-5 days. Single colony grown in the agar then was inoculated in to fresh agar plate. According to Yang et al. (2010) Scizochytrium Inoculation was done repeatedly until strains which could be isolated from macroalgae, fallen leaves of have characteristics of Schizochytrium has been mangrove trees and also mud from mangrove obtained. ecosystem. Schizochytrium grows in mangrove and coastal ecosystem. Several studies showed that For obtaining DNA sequences of 18S rRNA gene Schizochytrium is symbiosis with mud degrading from one strain, a single colony of the strain grown on bacteria, so that these microalgae also could be an agar plate was carefully transferred to a 50 mL isolated from soil or mud around mangroves. tube with 10 mL liquid medium containing 1 g/L peptone, 2 g/L yeast extract, and 4 g/L glucose Isolation techniques of Schizochytrium can be prepared with seawater. The culture was then cultivated initiated by taking samples from those places. The at 26°C for 1 week with continuous shaking (150 rpm). samples were stored in sterile plastic bags and sent The algal cells were collected by centrifugation (3.000 to the laboratory within 1 day for algal cell isolation. rpm for 5 min), rinsed with 5 mL deionized water, and Microalgal cells attached to samples were washed lyophilized before DNA sequencing (Yang et al., 2010). down using sterile seawater, passed through a 60- The resulting 18S rRNA gene sequences were then μm. plankton net to remove zooplankton,
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