Bioeng. Microbiol. Euglena gracilis has a pathway for the synthesis of EPA and subsequently DHA (Figure 2D), and the enzymes within this pathway have been characterised biochemically. Cell Res. 23, 12231228. 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). Phycol. Poultry Sci. There are a few isolated reports on genetic manipulation of E. gracilis. Br. Presently, only nutritious whole cell meal, the nutraceutical paramylon and cosmetics are commercially viable, while biofuel production from E. gracilis cell mass is in contrast not yet feasible. (2010). Structural and functional analysis of three beta-glucosidases from bacterium Clostridium cellulovorans, fungus Trichoderma reesei and termite Neotermes koshunensis. (2017). Biochim. doi: 10.1159/000343106, Ntefidou, M., Iseki, M., Watanabe, M., Lebert, M., and Hder, D.-P. (2003). The methylerythritol phosphate pathway contributes to carotenoid but not phytol biosynthesis in Euglena gracilis. Biochemistry 2, 11481154. Biochem. Biosci. 28, 423436. (1964). Enzymes of the FA biosynthesis pathway in E. gracilis have been shown to be crucial for the quality of WE derived from E. gracilis. J. Biol. (a) Culturing Euglena gracilis Klebs as batch cultures under 210 mol m-2 s-1; (b) Euglena gracilis Klebs growing under 210 mol m-2 s-1 at the exponential phase (Images by C.K. The -carotene content of E. gracilis cell mass grown in this two-step cultivation process or under PT conditions (3.4 or 3.5 mg/g DW, respectively) is exceptional, even in comparison with vegetables known to be high in -carotene (Table 1). Immobilization and characterization of E. gracilis extract with enriched laminaribiose phosphorylase activity for bienzymatic production of laminaribiose. J. Osaka Prefecture Ser. doi: 10.1111/j.1550-7408.2009.00452.x. 27, 140151. Trichoderma reesei, a superior cellulase source for industrial applications. Until now, the research on the health benefits of paramylon has focussed mainly on applications to treat human conditions, but it has been suggested that it also could be beneficial to livestock and fish health. Preliminary laboratory-scale experiments to determine E. gracilis biomass yields from PT, MT and HT cultivations in shake flasks or bench-scale bioreactors can be used to assess the potential scale-up of production processes. Euglena gracilis can accumulate large amounts of the reserve polysaccharide paramylon, a -1,3-glucan, which can constitute over 80% (w/w) of the dry weight (DW, biomass dried to a constant weight without oxidation) (Barsanti et al., 2001; Sun et al., 2018). doi: 10.1186/s13068-015-0264-5, Ogawa, Y., Noda, K., Kimura, S., Kitaoka, M., and Wada, M. (2014). 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. doi: 10.2174/0929867003374705, Papadaki, A., Mallouchos, A., Efthymiou, M.-N., Gardeli, C., Kopsahelis, N., Aguieiras, E.C.G., et al. doi: 10.1080/10408398.2016.1160362, Yadavalli, R., Rao, C.S., Rao, R.S., and Potumarthi, R. (2014). Another example is the production of self-assembling -1,3-glucan nanofibres derived from paramylon (Shibakami et al., 2013a). In the first step of this process, E. gracilis is cultivated under inexpensive PT conditions (Arashida and Mitra, 2015). Oxidative glucanases: cleavage of crystalline paramylon to make it accessible for other enzymes, possibly similar to oxidative cleavage of cellulose (Johansen, 2016). Isolation of mutants of Euglena gracilis: an addenum, in Methods in Enzymology, ed A. San Pietro (Cambridge: Academic Press), 2329. 30, 19451955. (1999). J. Biol. (2009). Biogas and bio-oil can also be produced from microalgal carbohydrates via thermochemical conversion processes like gasification at high temperatures or pyrolysis in the absence of oxygen, respectively (Behera et al., 2014). The in vitro synthesis of soluble -1,3-glucans could be an alternative to the hydrolysis approach. 93, 5571. Inui, H., Ishikawa, T., and Tamoi, M. (2017). doi: 10.7205/MILMED-D-14-00147. 16, 112. Firstly, by optimizing the solid-liquid ratio, extraction time, extraction . Univ. 142, 121126. Ultrastructural changes during carbon starvation in Euglena gracilis. doi: 10.1128/ec.00097-06. doi: 10.3382/ps/pey227. Hepatoprotective effects of paramylon, a -1,3-D-glucan isolated from Euglena gracilis Z, on acute liver injury induced by carbon tetrachloride in rats. Braz. Commun. (2017). To the best of our knowledge, there are currently no reports of a commercial -tocopherol production using E. gracilis. Lentinan with S-1 and paclitaxel for gastric cancer chemotherapy improve patient quality of life. Phytochemistry 116, 2127. The two novel studies proposed Euglena gracilis as a working model of gravitaxis [4], and the applied molecular toolkits and methods can be used to bioengineer E . However, further studies are needed to confirm this proposed regulatory role. 12, 8994. At pH values typical of AMD (pH 2.5-4) in the absence of elevated metals, E. gracilis outcompeted E. mutabilis (growth rates of 1.0 and 0.8 div d 1, respectively). Univ. MySQL. Best practices in heterotrophic high-cell-density microalgal processes: achievements, potential and possible limitations. Subcellular distribution of cobalamin-dependent methionine synthase in Euglena gracilis Z. Phytochemistry 35, 5961. Therefore, it has been widely applied in many fields, such as in food, supplements, and cosmetics (Gong et al. 64, 415419. Immunopathol. 426, 125134. (2001). For example, mice treated with 2,4,6-trinitrochlorobenzene, which would usually induce atopic dermatitis (AD)-like skin lesions, were fed with paramylon at 1% (w/w) of their diet. Paramylon also can act as a potent antioxidant protecting mice from acute hepatic injury induced by CCl4 treatment and was shown to alleviate non-alcoholic steatohepatitis in mice caused by a combination of streptozocin injection and high fat diet when orally administered at a dosage of 1 or 3 g/kg bodyweight (BW) per day, respectively (Sugiyama et al., 2009; Nakashima et al., 2018a). Nat. doi: 10.1046/j.1432-1033.2003.03364.x, Koizumi, N., Sakagami, H., Utsumi, A., Fujinaga, S., Takeda, M., Asano, K., et al. *Correspondence: Anwar Sunna, [email protected], Creative Commons Attribution License (CC BY). Euglena is a rare organism that combines both plant and animal cell characteristics to efficiently absorb nutrients. 207, 8390. Bioeng. Determination of the nutritive value of Euglena gracilis protein by in vitro digestion experiments and rat feeding tests. Despite substantial evidence for the health-enhancing bioactivities of paramylon and paramylon-derived compounds as outlined above, we are not aware of any clinical studies confirming these claims. 1, 15. Furthermore, there are effects attributed to paramylon beyond the immune response to pathogens. PLoS ONE 8, 16. For example, transformation of E. gracilis chloroplasts with the gene coding for a cyanobacterial fructose-1,6-/sedoheptulose-1,7-bisphosphatase has been achieved by biolistic bombardment (Doetsch et al., 2001; Ogawa et al., 2015). doi: 10.1016/j.phytochem.2015.05.010, Takeyama, H., Kanamaru, A., Yoshino, Y., Kakuta, H., Kawamura, Y., and Matsunaga, T. (1997). Immunomodulation and anti-cancer activity of polysaccharide-protein complexes. Department of Separation Science, LUT School of Engineering Science, LUT University, Finland, National Institute of Cardiology Ignacio Chavez, Mexico. 2019). PT cultivation on the other hand, although being a feasible alternative to HT cultivation, is restricted by low yields even after the technical optimisation of the cultivation process (Chae et al., 2006; Grimm et al., 2015; Hasan et al., 2017; Suzuki, 2017). Kataoka, H., Shimura, T., Mizoshita, T., Kubota, E., Mori, Y., Mizushima, T., et al. USDA National Nutrient Database for Standard Reference. (2012). Biotechnol. Fish Dis. (2018). 179 (Suppl. Biomass nutrient profiles of three microalgae: Spirulina platensis, Chlorella vulgaris, and Isochrisis galbana. BMC Genomics 17, 110. Chem. Bone Mineral Res. 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). For example, a multi-step cultivation process has been developed on a laboratory scale (in flasks) aiming to maximise WE synthesis. Multi-stage Process for Production of Immune Modulator. 53, 185190. In vivo 3, 243247. Ferment. J. Clin. 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). doi: 10.1271/bbb.80062, Ishikawa, T., Tajima, N., Nishikawa, H., Gao, Y., Rapolu, M., Shibata, H., et al. doi: 10.1292/jvms.09-0526, Sugiyama, A., Suzuki, K., Mitra, S., Arashida, R., Yoshida, E., Nakano, R., et al. BioSyst. Euglena Euglena, genus of more than 1,000 species of single-celled flagellated (i.e., having a whiplike appendage) microorganisms that feature both plant and animal characteristics. 4, 115. reported a nuclear transformation platform facilitating further molecular genetic studies and the metabolic modification of E. gracilis (Khatiwada et al., 2019). doi: 10.1002/elsc.200900077, antek, B., Friehs, K., Lotz, M., and Flaschel, E. (2012). J. Macromol. 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). doi: 10.1007/s11745-010-3395-2, Tokuoglu, ., and nal, M.K. Biochem. (2015b). Microscopic image of paramylon granules (B). Res. The granules are ~16 m long, surrounded by a biomembrane and show an unusual high degree of crystallinity, setting them apart from other carbohydrate storage products found in plants and algae (Miyatake and Kitaoka, 1983; Koizumi et al., 1993; Bumer et al., 2001; Barsanti et al., 2011; Monfils et al., 2011). (2017). 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). Chemical derivatives of paramylon (activated paramylon) have been shown to exhibit augmented or novel bioactivities. 5, 352362. Bioeng. They also act as a dietary fibre and have been shown to lower blood cholesterol levels (Nakashima et al., 2018b). 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