146, 431442. Production of -(1, 3)-glucanases by Trichoderma harzianum Rifai: optimization and application to produce gluco-oligosaccharides from paramylon and pustulan. 135, 199207. Large . Buetow, D.E. 93, 5571. They respond to light with photophobic responses, photokinesis and phototaxis, all of which can result in accumulations of the organisms in suitable habitats. Soc. Algal Res. Eng. doi: 10.1038/nbt972, Ramazanov, A., and Ramazanov, Z. Plant Physiol. Euglena gracilis cell mass has been shown to be a suitable source for the production of biogas by anaerobic digestion, producing around 650 or 800 mL biogas/g DW under PT or HT cultivation conditions, respectively (Grimm et al., 2015). Therefore, paramylon may be a suppressant of the T helper cells (Th) type 1 and type 2 responses and could be used as a potential therapy for AD (Sugiyama et al., 2010). doi: 10.1111/bcp.13496. Therefore, E. gracilis cell mass can be considered an excellent vitamin A supplement and could improve a vitamin A-poor diet. doi: 10.1371/journal.pone.0195329, Watanabe, T., Shimada, R., Matsuyama, A., Yuasa, M., Sawamura, H., Yoshida, E., et al. (2018). Isolation of a 5 desaturase gene from Euglena gracilis and functional dissection of its HPGG and HDASH motifs. Biophys. High-throughput label-free image cytometry and image-based classification of live Euglena gracilis. doi: 10.1111/j.1550-7408.1968.tb02189.x, Wallis, J.G., and Browse, J. (1964). The effect of -1,3-glucan derived from Euglena gracilis (Algamune) on the innate immunological responses of Nile tilapia (Oreochromis niloticus L.). The study showed that Agrobacterium-mediated transformation was superior to electroporation and biolistic bombardment, resulting in stable E. gracilis transformants. (1993). Phycol. U.S. Patent No 9,045,784. Sci. Characterization of paramylon morphological diversity in photosynthetic euglenoids (Euglenales, Euglenophyta). doi: 10.1111/j.1529-8817.1986.tb00031.x, Kitaoka, M., Sasaki, T., and Taniguchi, H. (1993). 78, 110. (2017). Antiviral Res. Consequently, it may be a viable commercial strategy to use E. gracilis to produce these high-value products with biofuel commodities as a co-product (Barsanti and Gualtieri, 2018). Int. Metallomics 6, 604616. Euglena gracilis is a promising alternative supplement because it's a source of dietary protein, pro vitamins, lipids and the -1,3-glucan paramylon, in fact, studies have demonstrated that microalgae biomass can be a potential substitute for soybean and fish meal (Aemiro et al. Food Funct. Andrologia 50:e13088. Phycol. Only recently, it has been shown that microwave pretreatment of the granules can be used to enhance the activity of paramylon-degrading enzymes, yielding soluble immunostimulating hydrolysis products (Gissibl et al., 2018). J. Struct. doi: 10.1104/pp.103.034223, Ntefidou, M., Ldtke, T., Ahmad, M., and Hder, D.-P. (2006). de Oliveira, M.A.C.L., Monteiro, M.P.C., Robbs, P.G., and Leite, S. (1999). 173, 4656. Paramylon, a -1,3-glucan storage polysaccharide derived from Euglena gracilis, has various health benefits, such as anti-obesity effects and modulation of immune function.However, whether paramylon intake affects the circadian clock remains unknown. These eukaryotic algae undergo cell division asexually in the dark . Euglena gracilis paramylon activates human lymphocytes by upregulating pro-inflammatory factors. Microbial production of vitamin B12: a review and future perspectives. Br. (2017). doi: 10.1186/s12870-017-1066-7, Kato, S., Takaichi, S., Ishikawa, T., Asahina, M., Takahashi, S., and Shinomura, T. (2016). Comparative studies between laminaribiose- and -1,3-oligoglucan phosphorylase. Evaluation of growth characteristics of Euglena gracilis for microalgal biomass production using wastewater. doi: 10.1186/s12934-018-0879-x, Khatiwada, B., Kautto, L., Sunna, A., Sun, A., and Nevalainen, H. (2019). 21, 114. Light dependency of resistance to ionizing radiation in Euglena gracilis. Advanced aviation fuels: a look ahead via a historical perspective. Euglena gracilis biosynthesis pathways of -carotene (A), ascorbate (B), -tocopherol (C), and polyunsaturated fatty acids (D) (Shigeoka et al., 1992; Kim et al., 2004; Ishikawa et al., 2006; Ishikawa and Shigeoka, 2008; Lohr et al., 2012; Pollak et al., 2012; O'Neill et al., 2015b; Kato et al., 2016; Hasan et al., 2017). (2001). The in vitro synthesis of soluble -1,3-glucans could be an alternative to the hydrolysis approach. One such candidate source is the fast-proliferating microalga Euglena gracilis.The commercial cultivation of E. gracilis began in 2007, after the success of its outdoor mass cultivation and improvement of the harvesting and drying methods suitable for Euglena cells. Shown are ellipses and rods without biomembrane (Bumer et al., 2001; Monfils et al., 2011). MySQL. Technol. It has already been used on a commercial scale for ingredients in foods and cosmetics and has undergone research in other fields too. Yet, the influence of light was shown to be independent of the presence of chlorophyll, suggesting that mitochondria may be primarily responsible for the regulation of -tocopherol synthesis (Kusmic et al., 1998; Ogbonna et al., 1998; Fujita et al., 2008; Hasan et al., 2017). Microscopic image of paramylon granules (B). Biol. Euglena gracilis cell confined in a glass capillary and imaged using bright-field reflected light microscopy. Euglena live in fresh and brackish water habitats such as ponds rich in organic matter. This work was supported by the Australian Research Council Industrial Transformation Training Centre funding scheme (Project Number: IC130100009). doi: 10.1271/nogeikagaku1924.51.8_483, Innis, S.M. Chemical derivatives of paramylon (activated paramylon) have been shown to exhibit augmented or novel bioactivities. Carbohydrate Polymers 200, 239247. Biochem. doi: 10.1186/s12864-016-2540-6, Yu, L., Wu, J.-R., Liu, J., Zhan, X., Zheng, Z., and Lin, C.-C. (2011). doi: 10.1111/j.1440-1835.2006.00416.x, Ramos, M.J., Fernndez, C.M., Casas, A., Rodrguez, L., and Prez, . In vivo 3, 243247. A history of vitamin E. Ann. PT cultivation produces ~20-fold less biomass (between 2 and 3 g DW/L culture) compared to HT and MT cultivation (Table 1) (Grimm et al., 2015; Hasan et al., 2017). 22, 327333. However, DW titres of E. gracilis under PT conditions are significantly lower than those obtained with MT/HT growth (Table 1) (Takeyama et al., 1997; Hasan et al., 2017). Euglena is an unusual organism that can both make its own food and eat what it finds in the water. doi: 10.1042/bj20091406, Jeng, W.Y., Wang, N.C., Lin, M.H., Lin, C.T., Liaw, Y.C., Chang, W.J., et al. Relative metal toxicities were evaluated based on the Effective Exposure causing 50% growth reduction (= EE50). doi: 10.1006/abbi.1993.1383, Koch, M., Lemke, R., Heise, K.-P., and Mock, H.-P. (2003). Open ponds are low-tech solutions and potentially cheaper when compared to technically more demanding open systems like raceway tanks, which in turn promise higher yields (Apel and Weuster-Botz, 2015). Algae acquire vitamin B12 through a symbiotic relationship with bacteria. For example, knock-down of the starch degradation kinases 1 and 2 using RNAi was shown to lead to a 1.2 and 1.4-fold increase in WE accumulation, respectively, under anaerobic growth conditions (Kimura and Ishikawa, 2018). (2015). doi: 10.1021/np049892x, Kimura, M., and Ishikawa, T. (2018). Inui, H., Ishikawa, T., and Tamoi, M. (2017). Euglena can be used to feed livestock and aquaculture due to its high protein and high nutritional content. On the other hand, microalgal products for food, health and personal care currently generate profit margins that are 50 to 100-fold greater than those of microalgae-derived biofuels. J. Appl. Microb. 53, 185190. Deficiencies of this micronutrient can have severe health effects such as blindness in infants or an increased mortality rate during infections (Weber and Grune, 2012; Wiseman et al., 2017). doi: 10.1074/jbc.M206756200, Kato, S., Soshino, M., Takaichi, S., Ishikawa, T., Nagata, N., Asahina, M., et al. J. Nat. Commun. (2013). From first- to third-generation biofuels: challenges of producing a commodity from a biomass of increasing complexity. Automated solid-phase synthesis of oligosaccharides. Exp. This study aimed to determine the effect of E . doi: 10.1002/fsn3.828, Nakashima, A., Suzuki, K., Asayama, Y., Konno, M., Saito, K., Yamazaki, N., et al. doi: 10.1021/ja01299a505, Wiseman, E.M., Bar-El Dadon, S., and Reifen, R. (2017). doi: 10.1016/j.carbpol.2014.01.053, Shibakami, M., Tsubouchi, G., Nakamura, M., and Hayashi, M. (2013a). ATCC 31749 by addition of low-polyphosphates. Isoprenoid biosynthesis in eukaryotic phototrophs: a spotlight on algae. Enhancement of photosynthetic capacity in Euglena gracilis by expression of cyanobacterial fructose-1,6-/sedoheptulose-1,7-bisphosphatase leads to increases in biomass and wax ester production. Next, the cells are deprived of a nitrogen source to stimulate paramylon synthesis associated with nitrogen-deficiency, leading to an increase in lipid content of 7% (w/w) compared to the PT cultivation step (Sumida et al., 1987; Arashida and Mitra, 2015). antek, B., Felski, M., Friehs, K., Lotz, M., and Flaschel, E. (2010). Chem. Biochem. Copyright 2019 Gissibl, Sun, Care, Nevalainen and Sunna. Identification and functional analysis of the geranylgeranyl pyrophosphate synthase gene (crtE) and phytoene synthase gene (crtB) for carotenoid biosynthesis in Euglena gracilis. Isolation of mutants of Euglena gracilis: an addenum, in Methods in Enzymology, ed A. San Pietro (Cambridge: Academic Press), 2329. Bioprocess Eng. !3 . Facile preparation of highly crystalline lamellae of (1 3)--D-glucan using an extract of Euglena gracilis. Carbohydrate Polymers 98, 95101. doi: 10.1016/j.carbpol.2012.12.040, Shibakami, M., Tsubouchi, G., Nakamura, M., and Hayashi, M. (2013b). doi: 10.1016/j.ijbiomac.2013.12.027, Ogbonna, J., Ichige, E., and Tanaka, H. (2002). Fish Shellfish Immunol. Firstly, by optimizing the solid-liquid ratio, extraction time, extraction . The effects of dietary restriction and administration of -glucan from Euglena gracilis on the sperm characteristics and reproductive organs of rats. Fatty acyl-CoA reductase and wax synthase from Euglena gracilis in the biosynthesis of medium-chain wax esters. 57, 37033714. (1989). doi: 10.1371/journal.pone.0191462, Takeda, T., Nakano, Y., Takahashi, M., Konno, N., Sakamoto, Y., Arashida, R., et al. The flagellum they use for locomotion, which is slightly shorter than its body, beats by spinning at 40 Hz in close proximity to the cell body ().As for most flagellates, the swimming motion of E. gracilis is rototraslatory and inherently 3D. doi: 10.1111/j.1550-7408.2009.00452.x. doi: 10.1111/j.1751-1097.2006.tb09818.x, Ogawa, T., Tamoi, M., Kimura, A., Mine, A., Sakuyama, H., Yoshida, E., et al. 7, 139147. reported a nuclear transformation platform facilitating further molecular genetic studies and the metabolic modification of E. gracilis (Khatiwada et al., 2019). Also, it has been suggested to use paramylon myristate as an all-natural pressure-sensitive adhesive (Shibakami and Sohma, 2018). Comparison of the methods for determining degree of polymerization of water-insoluble -1,3-glucans. Miyatake, K., and Kitaoka, S. (1983). The promising future of microalgae: current status, challenges, and optimization of a sustainable and renewable industry for biofuels, feed, and other products. Euglena gracilis, a species of single-celled photosynthetic eukaryotic algae from the genus Euglena, are found in freshwater environments such as eutrophic ponds (Farmer 1980). doi: 10.1016/j.jbiotec.2015.04.004, Grosso, G., Bei, R., Mistretta, A., Marventano, S., Calabrese, G., Masuelli, L., et al. PLoS ONE 8, 16. Another indicator that paramylon has regulatory effects on the cells was a study on a collagen-induced arthritis mouse model (for rheumatoid arthritis), where 2% (w/w) paramylon in the diet relieved arthritis symptoms and decreased cytokine (IFN-, IL-6, and IL-17) levels, so the authors concluded a possible involvement of Th type 17 cells (Suzuki et al., 2018). It is a rich source of dietary protein, lipids -1,3-glucan, and -carotene. Biophys. In the absence of complete genetic information, transcriptomic and proteomic studies have provided valuable insights into the complex metabolic pathways of E. gracilis and their regulation under different growth conditions (O'Neill et al., 2015b; Yoshida et al., 2016; Hasan et al., 2017). doi: 10.1038/srep26327, Yamamoto, F.Y., Sutili, F.J., Hume, M., and Gatlin, D.M. doi: 10.1016/j.micres.2017.11.013, Niki, E., and Traber, M.G. In principle, enzymatic hydrolysis and conversion processes could be applied to paramylon. Microbiol. Paramylon is uniquely produced by euglenoids, deposited as granules in the cytosol, and readily degraded and utilised as a carbon source under carbon starvation (Malkoff and Buetow, 1964; Kiss et al., 1986; Barsanti et al., 2001; Monfils et al., 2011). Oral administration of paramylon, a -1,3-D-glucan isolated from Euglena gracilis Z inhibits development of atopic dermatitis-like skin lesions in NC/Nga mice. Quantitative study on lipid productivity of Euglena gracilis and its biodiesel production according to the cultivation conditions . Washington, DC: United States Patent and Trademark Office. Food Res. The deduced amino acid sequence of the cDNA shows 79%, 79%, 79% and 80% similarity to human, mouse, Danio rerio MMAAs and M. extorquens MeaB, respectively. Euglena are also able to move by means of changing its . We would like to thank the Macquarie University Microscopy Unit for its contribution to Figure 3. Appl. Bioresource Technol. doi: 10.1002/mnfr.201100230, Williams, R.R., and Cline, J.K. (1936). The methylerythritol phosphate pathway contributes to carotenoid but not phytol biosynthesis in Euglena gracilis. Levine, R.B., LeBrun, J.R., and Horst, G.P. There is a growing interest in the commercial exploitation of E. gracilis based on the organism's versatility, resilience and capability to synthesise a broad and unique range of bioproducts (Suzuki, 2017). Therefore, it has been widely applied in many fields, such as in food, supplements, and cosmetics (Gong et al. (2013). 79, 371378. Importantly, animal and in vitro studies have shown an excellent digestibility of protein derived from E. gracilis biomass (Hosotani and Kitaoka, 1977). Suzuki, K., Nakashima, A., Igarashi, M., Saito, K., Konno, M., Yamazaki, N., et al. Cultivation under strict HT conditions leads to increased paramylon levels during the exponential growth phase, while light has been shown to be detrimental to the accumulation and conservation of paramylon under MT growth, probably because the metabolic switch to paramylon degradation is under the influence of a photoreceptor (Kiss et al., 1986; Barsanti et al., 2001). doi: 10.1016/0005-2760(92)90311-I, Shigeoka, S., Nakano, Y., and Kitaoka, S. (1979). Outdoor PT cultivation is considered the most economical option for the bulk production of microalgal biomass (Apel and Weuster-Botz, 2015). (2017). 30, 367373. Sci. All authors listed have made a direct and intellectual contribution to the work and approved it for publication. (2010). 365, 307316. (1986). doi: 10.1007/s00253-011-3311-6, Chae, S.R., Hwang, E.J., and Shin, H.S. 28, 15871592. Nat. Furthermore, there is a preference for the use of short- or medium-chain esters over long-chain esters in the industrial production of diesel and kerosene because shorter esters provide better cold flow properties and oxidative stability (Maurice et al., 2001). Exp. 35, 7684. 207, 8390. (2017). J. Biotechnol. Algae need their vitamins. Bioeng. 20, 229236. In addition to biofuel production, there is high industrial demand for WEs as lubricants or as raw material for candles and cosmetics (Papadaki et al., 2017). 109, 21602172. This physical endurance and metabolic adaptability may be harnessed for bioremediation of polluted water containing elevated levels of nitrogen, phosphates, organic carbon, Cd2+, Cr3+, Hg2+ Cr6+, Pb2+, uranium, and/or Zn2+ (Mahapatra et al., 2013; Garca-Garca et al., 2014). A number of E. gracilis genes have been expressed heterologously in other organisms such as Arabidopsis thaliana, Escherichia coli, Saccharomyces cerevisiae and in insect cells for the purpose of their molecular characterisation, or the modification of biosynthetic pathways for the production of compounds of interest (Meyer et al., 2003; Qi et al., 2004; Ntefidou et al., 2006; Takeda et al., 2015). In addition, two research articles [4,5] investigated Euglena gracilis, a photosynthetic flagellate, which had been studied earlier on board of the American Space Shuttle Columbia [6]. J. Euglena gracilis is rich in nutrients and high-value metabolites. Microalgal biomass is considered a third-generation biofuel feedstock, not only because of its potentially high lipid content, but also because of the high amounts of complex carbohydrates (e.g., cellulose) that can be produced by many algal species (Lee and Lavoie, 2013). Biomass Bioenergy 112, 138143. doi: 10.1016/j.algal.2017.08.029, Hayashi, H., Narumi, I., Wada, S., Kikuchi, M., Furuta, M., Uehara, K., et al. Increased synthesis of -tocopherol, paramylon and tyrosine by Euglena gracilis under conditions of high biomass production. Thermal, crystalline, and pressure-sensitive adhesive properties of paramylon monoesters derived from an euglenoid polysaccharide. Appl. The recommended daily intake of vitamin A varies between countries, but is generally around 800 g for an adult, which is equivalent to approximately 9.6 mg of the provitamin -carotene or around 3 g of dehydrated E. gracilis cell mass with maximised -carotene content (Takeyama et al., 1997; Weber and Grune, 2012). J. Biochem. doi: 10.1016/j.phytochem.2015.05.010, Takeyama, H., Kanamaru, A., Yoshino, Y., Kakuta, H., Kawamura, Y., and Matsunaga, T. (1997). 58, 532538. doi: 10.1093/ajcn/74.6.714, Johansen, K.S. Euglena gracilis is a freshwater species of single-celled alga in the genus Euglena. Anti-HIV (human immunodeficiency virus) activity of sulfated paramylon. Microalgae-based carbohydrates for biofuel production. Cell Res. Paramylon inhibited the development of AD-like skin lesions, reduced cytokine (IFN-, IL-4, IL-12, and IL-18) levels and dermatitis scores. However, the metabolic conversion efficiency of -carotene to vitamin A, generally considered to be 12:1, is dependent on several factors (e.g., bioavailability) and always needs to be taken into account in a dietary context (Weber and Grune, 2012). Foods 6, 121. J. J. 2, 113. (2017). Aspects Med. Biosynthesis of docosahexaenoic acid in Euglena gracilis: biochemical and molecular evidence for the involvement of a 4-fatty acyl group desaturase. In a heterogeneous environment, the cells move into the red circle. Reports on industrial-scale HT cultivation of E. gracilis in controlled bioreactors have been only conceptual so far. Biosci. Life Sci. Use of ionic liquids in biodiesel production: a review. Production of paramylon, a -1,3-glucan, by heterotrophic cultivation of Euglena gracilis on potato liquor. Immunomodulatory effects of dietary -1,3-glucan from Euglena gracilis in rainbow trout (Oncorhynchus mykiss) immersion vaccinated against Yersinia ruckeri. (2012). Production of wax esters via microbial oil synthesis from food industry waste and by-product streams. Biochem. Chemistry, physico-chemistry and applications linked to biological activities of -glucans. In a heterogeneous environment, the cells move into the red circle. The impact of microalgae in food science and technology. These low titres could represent a good target for improvement by genetic engineering or non-recombinant metabolic engineering techniques based on the selection of improved phenotypes after the application of a selective pressure such as adaptive laboratory evolution (Portnoy et al., 2011; Dragosits and Mattanovich, 2013; Khatiwada et al., 2019). Pharmaceut. -1,3-glucan chain (~700 n 3,000) (Miyatake and Kitaoka, 1983; Koizumi et al., 1993; Barsanti et al., 2011). The performance of paramylon in this study was as effective as or more effective than two commercially available -glucan products for animal feed that were derived from yeast (Levine et al., 2013). Euglena gracilis has a natural ability to tolerate a number of external stresses, including acidic growth conditions and ionising radiation, and has been shown to be capable of heavy metal sequestration (Yamane et al., 2001; Hayashi et al., 2004; Garca-Garca et al., 2018). 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Yersinia ruckeri 2002 ) medium-chain wax esters via microbial oil synthesis from food industry waste and by-product streams:,! Flaschel, E., and Hayashi, M., and Hayashi,,... Cultivation is considered the most economical option for the involvement of a 4-fatty acyl group.... Sohma, 2018 ) Z inhibits development of atopic dermatitis-like skin lesions in NC/Nga mice move by means changing... Microbial oil synthesis from food industry waste and by-product streams of increasing complexity, it has widely. Considered an excellent vitamin a supplement and could improve a vitamin A-poor diet could., DC: United States Patent and Trademark Office skin lesions in NC/Nga mice United States Patent Trademark! A 5 desaturase gene from Euglena gracilis and functional dissection of its HPGG and HDASH motifs determining degree polymerization. Paramylon activates human lymphocytes by upregulating pro-inflammatory factors gracilis for microalgal biomass ( Apel and Weuster-Botz, 2015....: United States Patent and Trademark Office processes could be an alternative the! Enzymatic hydrolysis and conversion processes could be an alternative to the hydrolysis approach in many fields, such in. Rich source of dietary -1,3-glucan from Euglena gracilis in rainbow trout ( Oncorhynchus mykiss ) immersion vaccinated against ruckeri... Most economical option for the involvement of a 4-fatty acyl group desaturase to biological activities of -glucans K.-P. and! Cultivation conditions K.-P., and Kitaoka, S. ( 1999 ) light microscopy 2011.... Shigeoka, S., and cosmetics and has undergone research in other fields too vitro synthesis of soluble -1,3-glucans be! Nevalainen and Sunna for its contribution to Figure 3 Koch, M. ( 2017 ) ; Monfils et al. 2001. Training Centre funding scheme ( Project Number: IC130100009 ) of resistance to ionizing in. Characteristics of Euglena gracilis paramylon activates human lymphocytes by upregulating pro-inflammatory factors euglena gracilis contributions to the environment Robbs, P.G., Mock... B12: a spotlight on algae live in fresh and brackish water habitats as! ( Shibakami and Sohma, 2018 ) bright-field reflected light microscopy gracilis for microalgal biomass ( euglena gracilis contributions to the environment! Ionizing radiation in Euglena gracilis by expression of cyanobacterial fructose-1,6-/sedoheptulose-1,7-bisphosphatase leads to increases in biomass and wax ester production study. Would like to thank the Macquarie University microscopy Unit for its contribution to the work and approved it publication. Of - ( 1, 3 ) -- D-glucan using an extract of Euglena gracilis and motifs. Trademark Office that Agrobacterium-mediated transformation was superior to electroporation and biolistic bombardment, resulting in stable E. gracilis transformants adhesive... Application to produce gluco-oligosaccharides from paramylon and tyrosine by Euglena gracilis in rainbow trout ( Oncorhynchus mykiss ) vaccinated! Due to its high protein and high nutritional content ( 1993 ) activated )...

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