At present, only two enzymes of this pathway, the D-galacturonate reductase and aldonolactonase, have been biochemically characterised in vitro. E. gracilis has been a popular model organism and of considerable biotechnological interest, but the absence of a gene catalogue has hampered both basic research and translational efforts. Heterologous expression of photoactivated adenylyl cyclase (PAC) genes from the flagellate Euglena gracilis in insect cells. doi: 10.1016/j.jhazmat.2017.09.008, Giese, E., Dekker, R., Barbosa, A., da Silva, M., and da Silva, R. (2011). Vogel, K., and Barber, A.A. (1968). Large-scale cultivation of Euglena, in Euglena: Biochemistry, Cell and Molecular Biology, eds S. D. Schwartzbach and S. Shigeoka (Cham: Springer International Publishing), 285293. Food Sci. 38, 9951008. (2001). In addition, a film dressing prepared from paramylon was shown to speed up wound healing in mice, possibly by regulating the immune response (Yasuda et al., 2018). (2017). PT cultivation of E. gracilis is currently favoured, probably due to lower cost and relative ease of set-up, but HT cultivation for higher yields of cell mass and paramylon could soon become reality. Abstract Motile microorganisms such as the green Euglena gracilis use a number of external stimuli to orient in their environment. Buetow, D.E. Euglena gracilis shows an absolute requirement for vitamins B1 (thiamine) and B12 (cobalamin), so complex E. gracilis media usually contain at least 50 ng/L and 10 ng/L, respectively, of these compounds (Isegawa et al., 1987; Shigeoka et al., 1987; Croft et al., 2005; Hasan et al., 2017). J. Appl. bioRxiv 2017, 159. Biophys. doi: 10.1016/j.carbpol.2013.05.026, Shigeoka, S., Ishiko, H., Nakano, Y., and Mitsunaga, T. (1992). Department of Separation Science, LUT School of Engineering Science, LUT University, Finland, National Institute of Cardiology Ignacio Chavez, Mexico. doi: 10.3382/ps/pey227. doi: 10.2216/09-112.1, Mller, C., Ortmann, T., Abi, A., Hartig, D., Scholl, S., and Jrdening, H.-J. (1967). (2012). doi: 10.1046/j.1432-1033.2003.03364.x, Koizumi, N., Sakagami, H., Utsumi, A., Fujinaga, S., Takeda, M., Asano, K., et al. For example, introduction of acyl groups with different alkyl chain length into paramylon molecules has been shown to produce an alternative to petroleum-based resins (Shibakami et al., 2014). Euglena cells contain bio-compounds like Paramylon (-glucan) which enhances the immune system, helps remove undesirable substances like fats and cholesterol, and also reduces the level of uric acid in the blood. doi: 10.1016/j.plantsci.2011.07.018, Mahapatra, D.M., Chanakya, H.N., and Ramachandra, T.V. (2018). J. Hepatogastroenterology 56, 547550. Hazard. Mol. saccharophila. However, it has also been shown that -tocopherol titres per g DW decrease under MT/HT cultivation (without ethanol) over time (Grimm et al., 2015; Hasan et al., 2017). There are several health benefits associated with -glucans, including immunostimulatory and antioxidant effects (Barsanti et al., 2011). As a result, corneal epithelial cell migration was increased and the acute inflammatory reaction caused by corneal alkali burns in vivo was supressed (Choi et al., 2013). doi: 10.1016/0005-2760(92)90311-I, Shigeoka, S., Nakano, Y., and Kitaoka, S. (1979). There are strong indications that this pretreatment of paramylon has the effect of an increase in stimulation of leucocytes and hepatoprotection, probably because the -1,3-glucan triple-helices of paramylon are disrupted in the process and the apparently more bioactive single helices are thereby exposed (Kataoka et al., 2002; Kusmic et al., 2018). Sci. Conversion of biowaste leachate to valuable biomass and lipids in mixed cultures of Euglena gracilis and chlorophytes. Technological trends, global market, and challenges of bio-ethanol production. This work was supported by the Australian Research Council Industrial Transformation Training Centre funding scheme (Project Number: IC130100009). Advertisement randellpierre23 Answer: However, carbohydrate and lipid yields in E. gracilis can be improved by new cultivation technologies and metabolic engineering approaches, possibly providing an affordable alternative to fossil fuels in the near future. One of the highest paramylon titres reported (16 g/L culture) was obtained in a repeated-batch cultivation under HT conditions in the dark (Table 1), using a medium supplemented with potato liquor, vitamins, and a high concentration of glucose (30 g/L) (antek et al., 2012; Grimm et al., 2015). MySQL. Biol. 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). Shibakami, M., Shibata, K., Akashi, A., Onaka, N., Takezaki, J., Tsubouchi, G., et al. Eng. However, there is no further information on whether this proposed system was ever built and tested. The 8-desaturase of Euglena gracilis: an alternate pathway for synthesis of 20-carbon polyunsaturated fatty acids. doi: 10.1007/s11095-018-2553-8, Shibakami, M., and Sohma, M. (2018). 9, 368373. J. Biol. 84, 659672. and their products currently on the market. (2001). Engineering solutions for open microalgae mass cultivation and realistic indoor simulation of outdoor environments. (1968). 41, 15791588. J. Lipid Res. 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. Int. (2017). This approach led to the isolation of a new strain capable of accumulating lipids to up to 0.2 g/g DW under hypoxic cultivation conditions, which represents a 40% (w/w) increase in lipid content compared to the original strain (Yamada et al., 2016). Suzuki, K., Nakashima, A., Igarashi, M., Saito, K., Konno, M., Yamazaki, N., et al. (2006). As yet, a maximum total lipid and WE titre of ~0.7 and 0.6 g/g DW, respectively, was reached during anaerobic cultivation of a natural E. gracilis isolate with the addition of an elongase inhibitor (Tucci et al., 2010). Bone Mineral Res. The alga Euglena gracilis (E. gracilis) has recently gained attention as a health food, but its effects on human gut microbiota remain unknown. Immunomodulation and anti-cancer activity of polysaccharide-protein complexes. Am. PLoS ONE 8, 16. Determination of the nutritive value of Euglena gracilis protein by in vitro digestion experiments and rat feeding tests. Paramylon is the water-insoluble storage polysaccharide of E. gracilis, which consists of -1,3-linked glucose subunits, and has an estimated molecular weight between 100 and 500 kDa (Figure 3A) (Miyatake and Kitaoka, 1983; Koizumi et al., 1993; Barsanti et al., 2011). Isoprenoid biosynthesis in eukaryotic phototrophs: a spotlight on algae. doi: 10.1016/S0031-9422(00)85672-1, Meyer, A., Cirpus, P., Ott, C., Schlecker, R., Zhringer, U., and Heinz, E. (2003). Eng. Eng. Mouse and rat studies, as well as experiments with different mammalian cell lines have been performed to determine the potential impact of the -1,3-glucan paramylon on human health. Chem. While some microalgae may achieve higher total lipid contents (Table 1), E. gracilis has the competitive advantage of a better WE/total lipid ratio of up to 0.8 g/g, as lipids produced by most microalgae are typically FAs (Maxwell et al., 1968; Tucci et al., 2010; Matos, 2017; Khan et al., 2018). doi: 10.1364/BOE.7.002703, Levine, R., Horst, G., Tonda, R., Lumpkins, B., and Mathis, G. (2018). 2019). doi: 10.1186/s12864-016-2540-6, Yu, L., Wu, J.-R., Liu, J., Zhan, X., Zheng, Z., and Lin, C.-C. (2011). Importantly, animal and in vitro studies have shown an excellent digestibility of protein derived from E. gracilis biomass (Hosotani and Kitaoka, 1977). doi: 10.1007/s12257-010-0145-5, Zhu, G., Ou, Q., Zhang, T., Jiang, X., Sun, G., Zhang, N., et al. Technol. 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). Microbiol. J. Appl. Production of -(1, 3)-glucanases by Trichoderma harzianum Rifai: optimization and application to produce gluco-oligosaccharides from paramylon and pustulan. The enzymatic conversion of major algal and cyanobacterial carbohydrates to bioethanol. Chemical modification potentiates paramylon induction of antimicrobial activity. doi: 10.1111/jpy.12758, Sutivisedsak, N., Leathers, T.D., Bischoff, K.M., Nunnally, M.S., and Peterson, S.W. (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. doi: 10.1016/0005-2744(69)90253-8, Barsanti, L., Bastianini, A., Passarelli, V., Tredici, M.R., and Gualtieri, P. (2000). Curr. doi: 10.1271/bbb.80062, Ishikawa, T., Tajima, N., Nishikawa, H., Gao, Y., Rapolu, M., Shibata, H., et al. Anal. 365, 307316. Food Chem. doi: 10.1016/j.algal.2018.02.001, Barsanti, L., Passarelli, V., Evangelista, V., Frassanito, A.M., and Gualtieri, P. (2011). doi: 10.1023/a:1008011201437, O'Neill, E.C., Trick, M., Henrissat, B., and Field, R.A. (2015a). Eng. doi: 10.1021/np049892x, Kimura, M., and Ishikawa, T. (2018). Nippon Nogeikagaku Kaishi 51, 483488. 72, 11431154. (2018). Cell Factories 16, 113. Biomass production in mixotrophic culture of Euglena gracilis under acidic condition and its growth energetics. From these, paramylon is already marketed as an immunostimulatory agent in nutraceuticals. Automated solid-phase synthesis of oligosaccharides. doi: 10.1080/17597269.2016.1192444, Khan, M.I., Shin, J.H., and Kim, J.D. 7, 139147. Two alternative metabolic routes to -carotene have been identified biochemically or via transcriptomics in E. gracilis: the methylerythritol phosphate and the mevalonate pathway (Disch et al., 1998; Kim et al., 2004; O'Neill et al., 2015b). (2012). HT cultivation with glucose as a carbon source in shake flasks usually reaches DW titres of ~12 g/L culture, whereas repeated-batch cultivation supplemented with potato liquor and glucose in a bioreactor (5 L working volume) was shown to reach ~23 g/L culture (antek et al., 2012; Grimm et al., 2015; Hasan et al., 2017). doi: 10.2527/af.2013-0010, Lei, C., Ito, T., Ugawa, M., Nozawa, T., Iwata, O., Maki, M., et al. Paramylon and other -1,3-glucans are of special interest because of their reported immunostimulatory and antimicrobial bioactivities (Kiss et al., 1986; Barsanti et al., 2001; Russo et al., 2017; Gissibl et al., 2018). AG was supported by an international Macquarie University Research Excellence Scholarship. The technical set-up of a cultivation also plays an important role in -tocopherol production, for example, hydrodynamic stress caused by fast stirring with baffled plates in a bioreactor cultivation has been shown to have a detrimental effect on the final yields of -tocopherol during HT cultivation (Ogbonna et al., 1998). (2010). Carotenoids: biochemistry, pharmacology and treatment. The unique ascorbate synthesis pathway from E. gracilis (Figure 2B) has been elucidated using radiotracer experiments, as well as transcriptomic and proteomic studies (Ishikawa and Shigeoka, 2008; O'Neill et al., 2015b; Hasan et al., 2017). Express 7, 27032708. Eur. Euglena gracilis is a common primary producer in aquatic ecosystems where it produces a large biomass with beneficial metabolites that have a high commercial value. , T.V Institute of Cardiology Ignacio Chavez, Mexico Sohma, M., and challenges bio-ethanol! 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