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. Biotechnol. In contrast, some vegetable oils (e.g., olive, sunflower, and wheat germ oil) are marketed as good sources of daily vitamin E intake, but only have maximum -tocopherol contents of 1.5 mg/g (Table 1), which further emphasises the commercial potential of E. gracilis as a viable candidate for industrial -tocopherol production (Psomiadou et al., 2000; US Department of Agriculture., 2018). 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). (2010). Curr. Firstly, by optimizing the solid-liquid ratio, extraction time, extraction . 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: 10.1016/j.jplph.2004.04.005, Henchion, M., Hayes, M., Mullen, A., Fenelon, M., and Tiwari, B. 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. World Intellectual Property Organization. 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). However, there is no further information on whether this proposed system was ever built and tested. doi: 10.1074/jbc.M206756200, Kato, S., Soshino, M., Takaichi, S., Ishikawa, T., Nagata, N., Asahina, M., et al. 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: 10.1016/0005-2744(67)90227-6, Matos, .P. (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: 10.1271/bbb.60327, Ishikawa, T., and Shigeoka, S. (2008). Biological, and Environmental Sciences, Faculty of Agriculture, Tottori . (2018). Eukaryot. The majority of the E. gracilis-derived lipids (WEs and FAs) are suitable for biodiesel production because they are saturated or have a low degree of unsaturation (Rosenberg, 1963; Tucci et al., 2010; Furuhashi et al., 2015). 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). However, unlike higher plants, E. gracilis produces almost exclusively the -form of tocopherol (Shigeoka et al., 1986; O'Neill et al., 2015b; Fritsche et al., 2017; Hasan et al., 2017). In a similar study, mice were provided with a diet containing lower than 1% (w/w) paramylon and challenged with a potentially lethal dose of E. coli. (2017). 91, 3146. Acta (BBA) Enzymol. doi: 10.1038/s41598-017-14077-6, Torihara, K., and Kishimoto, N. (2015). doi: 10.1002/elsc.200900077, antek, B., Friehs, K., Lotz, M., and Flaschel, E. (2012). 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). 2, 113. J. Biol. doi: 10.1271/bbb.80062, Ishikawa, T., Tajima, N., Nishikawa, H., Gao, Y., Rapolu, M., Shibata, H., et al. Biochem. They possess two flagellae, one long, one short, which can allow the organisms to move. Biosci. doi: 10.1042/bj3330381, Doetsch, N.A., Favreau, M.R., Kuscuoglu, N., Thompson, M.D., and Hallick, R.B. Several reports have described the successful mutagenesis (e.g., by irradiation) of E. gracilis (Schiff et al., 1980; Yamada et al., 2016; Suzuki, 2017). A putative enzymatic mechanism for complete synergistic hydrolysis of the polysaccharide paramylon to glucose is shown in Figure 3C. Food Sci. 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). Only central enzymes/substrates/intermediates/products are shown. Rep. 7, 113. 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. However, paramylon granules have been shown to be recalcitrant to enzymatic degradation and it is very likely that a consortium of different enzymes is required for the efficient degradation of this polysaccharide (Sutivisedsak et al., 2013). Appl. doi: 10.1111/j.1751-1097.2006.tb09818.x, Ogawa, T., Tamoi, M., Kimura, A., Mine, A., Sakuyama, H., Yoshida, E., et al. Plant Physiol. Microalgae can accumulate large quantities of proteins intracellularly and therefore represent an attractive alternative to more traditional sources of dietary protein like meat and fish (Henchion et al., 2017; Ritala et al., 2017). doi: 10.1006/abbi.1999.1167, Wang, Y., Seppnen-Laakso, T., Rischer, H., and Wiebe, M.G. doi: 10.1039/C0NP00018C, Barsanti, L., Vismara, R., Passarelli, V., and Gualtieri, P. (2001). 33, 111120. doi: 10.1111/j.1365-2672.2010.04848.x, Rosenberg, A. 173, 4656. (2001). Relative metal toxicities were evaluated based on the Effective Exposure causing 50% growth reduction (= EE50). -glucan in foods and its physiological functions. It is related to plants, animals and fungi - all known as eukaryotes - but is a separate . Microb. Fatty acid content in wild type and WZSL mutant of Euglena gracilis. Production of paramylon, a -1,3-glucan, by heterotrophic cultivation of Euglena gracilis on potato liquor. Biotechnol. Life Sci. doi: 10.1128/ec.00097-06. 182, 197215. doi: 10.1016/j.jsb.2010.07.008, Jiang, Q., Christen, S., Shigenaga, M.K., and Ames, B.N. Euglena in time: evolution, control of central metabolic processes and multi-domain proteins in carbohydrate and natural product biochemistry. These findings highlight the potential of E. gracilis enzymes for the microbial production of biofuels (Teerawanichpan and Qiu, 2010; Tomiyama et al., 2017; Nandy and Srivastava, 2018). doi: 10.3390/foods6070053, Hosotani, K., and Kitaoka, S. (1977). Curr. The contribution of -carotene to vitamin A supply of humans. 31, 107115. Eur. In addition, the enzyme ascorbate peroxidase reduces excess hydrogen peroxide to water by oxidation of ascorbate (Shigeoka et al., 1980; Ishikawa et al., 2010; Hasan et al., 2017). 23, 12231228. Initial transcriptomic studies indicated that amino acid synthesis pathways in E. gracilis were similar to those used by plants and bacteria (O'Neill et al., 2015b). doi: 10.1038/srep26327, Yamamoto, F.Y., Sutili, F.J., Hume, M., and Gatlin, D.M. Vet. Biotechnol. saccharophila. J. Appl. Moreover, E. gracilis WEs are more suited for catalytic cracking for the production of fuel as the reaction is faster with less formation of unwanted poorly combustible polyaromatic compounds (Shimada et al., 2018). 11), 8287. 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). Growth and chemical composition of Spirulina maxima and Spirulina platensis biomass at different temperatures. 35, 7684. (2016). Lipids 47, 913926. In the Western world, major health conditions like cardiovascular and neurological diseases have been linked directly to a lack of long-chain -3 PUFAs, especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), in the dietary intake (Deckelbaum and Torrejon, 2012; Innis, 2014). Am. Biosci. The 8-desaturase of Euglena gracilis: an alternate pathway for synthesis of 20-carbon polyunsaturated fatty acids. (2018a). Nevertheless, ascorbate and other bioproducts like vitamin E remain an important component of commercially available food supplements based on E. gracilis cell mass (Barsanti and Gualtieri, 2018). Determination of the nutritive value of Euglena gracilis protein by in vitro digestion experiments and rat feeding tests. Yet, it is assumed that the conversion of -tocopherol to -tocopherol is the rate-limiting step in the pathway of plants (Fritsche et al., 2017). Plant Cell Physiol. Suzuki, K., Nakashima, A., Igarashi, M., Saito, K., Konno, M., Yamazaki, N., et al. Phycol. 109, 21602172. Euglena sp. Production of paramylon, a -1,3-glucan, by heterotrophic growth of Euglena gracilis on potato liquor in fed-batch and repeated-batch mode of cultivation. doi: 10.1016/j.pisc.2015.07.002, O'Neill, E.C., Trick, M., Hill, L., Rejzek, M., Dusi, R.G., Hamilton, C.J., et al. The two predominant options for the mass cultivation of microalgae are: (i) closed (photo)bioreactors; and (ii) open systems exposed to the environment. Front. 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). Am. (1967). 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). (2018b). !3 . Antitumor activity of the -glucan paramylon from Euglena against preneoplastic colonic aberrant crypt foci in mice. doi: 10.1016/j.algal.2018.08.007, Traber, M.G. 11, 28082820. (2015). doi: 10.1007/s00253-008-1443-0, Furuhashi, T., Ogawa, T., Nakai, R., Nakazawa, M., Okazawa, A., Padermschoke, A., et al. Lipids 50, 483492. Biotechnol. However, the product (laminaribiose) yield of the immobilisation approach in combination with the SP enzyme was just 20% (w/w) (Mller et al., 2016). have proposed a multi-stage process where smaller bioreactors generate the inocula for the bigger stages (up to 1,000,000 L) with the aim to increase biomass titres to 120 g/L culture or higher (Levine et al., 2013). (2001). Chloroplast transformation in Euglena gracilis: splicing of a group III twintron transcribed from a transgenic psbK operon. A comprehensive assessment of the biosynthetic pathways of ascorbate, -tocopherol and free amino acids in Euglena gracilis var. doi: 10.1016/j.fsi.2018.04.003, Yamane, Y.-,I., Utsunomiya, T., Watanabe, M., and Sasaki, K. (2001). Advertisement randellpierre23 Answer: doi: 10.1039/C3MT00313B, Garca-Garca, J.D., Pea-Sanabria, K.A., Snchez-Thomas, R., and Moreno-Snchez, R. (2018). Nippon Nogeikagaku Kaishi 51, 483488. Improvement of the performance of E. gracilis has mainly relied on developing cultivation conditions to favour the synthesis of the compound of interest, followed by scale-up of the cultivation volume, because of a general lack of genetic information on the metabolic pathways leading to the various bioproducts (O'Neill et al., 2015a; Wang et al., 2018). Comparative assessment of the Euglena gracilis var. However, an in-depth proteomic analysis recently revealed a unique pathway for arginine biosynthesis in E. gracilis, which is independent from the urea cycle commonly utilised by other eukaryotic organisms (Hasan et al., 2017). doi: 10.1007/s002940000174, Dragosits, M., and Mattanovich, D. (2013). 7:108. doi: 10.3389/fbioe.2019.00108. FEBS Lett. 22, 327333. J. Appl. Carbohydrate Polymers 105, 9096. Braz. It is likely that certain disadvantages of HT cultivation prevented its realisation such as the cost of the medium containing an organic carbon source like glucose (antek et al., 2010). Eng. doi: 10.1039/C3FO60256G, Weber, D., and Grune, T. (2012). (2016). Adaptive laboratory evolution - principles and applications for biotechnology. J. doi: 10.3177/jnsv.64.8, Nakazawa, M., Andoh, H., Koyama, K., Watanabe, Y., Nakai, T., Ueda, M., et al. Nutr. (2012). Variability of wax ester fermentation in natural and bleached Euglena gracilis strains in response to oxygen and the elongase inhibitor flufenacet. Reported EPA and DHA titres in E. gracilis are negligible (Table 1) and apparently independent of cultivation conditions (light/dark cultivation) (Korn, 1964; Barsanti et al., 2000). (2004). Vitamin-dependence is common amongst microalgae, but rarely discussed in the context of the scale-up of E. gracilis cultivation (Croft et al., 2006). Production of wax esters via microbial oil synthesis from food industry waste and by-product streams. Scope of algae as third generation biofuels. doi: 10.1364/BOE.7.002703, Levine, R., Horst, G., Tonda, R., Lumpkins, B., and Mathis, G. (2018). Therefore, E. gracilis cell mass can be considered an excellent vitamin A supplement and could improve a vitamin A-poor diet. Biotechnol. Lett. Consequently, -carotene titres per g DW are highest under PT cultivation conditions (Table 1). Photosynthesis of Euglena gracilis under cobalamin-sufficient and -limited growing conditions. Biomass nutrient profiles of three microalgae: Spirulina platensis, Chlorella vulgaris, and Isochrisis galbana. doi: 10.1186/1475-2859-12-64, Ebenezer, T.E., Zoltner, M., Burrel, A., Nenarokova, A., Anna, Vanclov, M.G.N., et al. Fatty acyl-CoA reductase and wax synthase from Euglena gracilis in the biosynthesis of medium-chain wax esters. Delta-9 Elongases and Their Use in Making Polyunsaturated Fatty Acids. 270, 8492. Biotechnol. Paramylon synthesis and chloroplast structure associated with nutrient levels in Euglena (Euglenophyceae). 135, 199207. Colonic aberrant crypt foci in mice g DW are highest under PT cultivation conditions ( Table 1.... Spirulina platensis biomass at different temperatures to move and bleached Euglena gracilis under cobalamin-sufficient and -limited growing.. Aberrant crypt foci in mice H., and Isochrisis galbana the polysaccharide paramylon to glucose is shown Figure! Friehs, K., and Flaschel, E. ( 2012 ) % growth reduction ( = EE50 ) Torihara. 2001 ) proteins in carbohydrate and natural product biochemistry putative enzymatic mechanism complete... Preneoplastic colonic aberrant crypt foci in mice for complete synergistic hydrolysis of the nutritive value of Euglena gracilis,! Seppnen-Laakso, T., Rischer, H., and Kitaoka, S. 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