doi: 10.1007/s002940000174, Dragosits, M., and Mattanovich, D. (2013). Eng. (2018). Alternatively, conventional mutagenesis could be used to generate 'natural' mutant strains with desired traits, a process which is generally not considered GM. 54, 529538. Bioproducts from E. gracilis can be produced under various cultivation conditions discussed in this review, and their yields are relatively high when compared with those achieved in microalgal systems. 61, 207212. Life Sci. Biochim. Algal Res. 54, 255259. Bioresource Technol. Rounding of the cell body, as determined by the reconfigurations of the pellicle . It has a highly flexible cell surface, allowing it to change shape from a thin cell up to 100 m long, to a sphere of approximately 20 m. doi: 10.1007/s11745-010-3395-2, Tokuoglu, ., and nal, M.K. It is a rich source of dietary protein, lipids -1,3-glucan, and -carotene. Euglena, especially Euglena gracilis, is the most studied member of the Euglenaceae. At present, only two enzymes of this pathway, the D-galacturonate reductase and aldonolactonase, have been biochemically characterised in vitro. Abstract Motile microorganisms such as the green Euglena gracilis use a number of external stimuli to orient in their environment. Chemistry, physico-chemistry and applications linked to biological activities of -glucans. 293, 28652876. Biochem. Vitamin-dependence is common amongst microalgae, but rarely discussed in the context of the scale-up of E. gracilis cultivation (Croft et al., 2006). Biosci. 16, 112. J. Biol. doi: 10.4172/2167-7972.1000102, Gissibl, A., Care, A., Parker, L.M., Iqbal, S., Hobba, G., Nevalainen, H., et al. This study aimed to determine the effect of E . Biochem. J. Protozool. A solution to this problem could be the genetic modification (GM) of E. gracilis to improve biomass and/or bioproduct yields. Vitamin E inadequacy in humans: causes and consequences. doi: 10.3945/jn.111.148080, Disch, A., Schwender, J., Mller, C., Lichtenthaler, H.K., and Rohmer, M. (1998). Comparable levels of 40 mg/L culture or 3.7 mg/g DW were reached after a relatively short incubation time of 120 h using a bleached strain in HT batch cultivation with a medium supplemented with ethanol, glutamate and malate (Rodrguez-Zavala et al., 2010). Foods 6, 121. doi: 10.1080/10408398.2016.1160362, Yadavalli, R., Rao, C.S., Rao, R.S., and Potumarthi, R. (2014). Accordingly, the supplementation of both vitamins, produced either by chemical synthesis (thiamine) or extracted from bacterial sources (cobalamin), would be a factor in a scaled-up E. gracilis cultivation and contributing to the overall costs of the process (Williams and Cline, 1936; Fang et al., 2017). Biotechnol. Biochem. Anim. Fish Shellfish Immunol. J. Appl. No use, distribution or reproduction is permitted which does not comply with these terms. doi: 10.1111/j.1529-8817.1986.tb00031.x, Kitaoka, M., Sasaki, T., and Taniguchi, H. (1993). 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). Wax ester fermentation and its application for biofuel production, in Euglena: Biochemistry, Cell and Molecular Biology, eds S. D. Schwartzbach and S. Shigeoka (Cham: Springer International Publishing), 269283. Fuel 80, 747756. Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting. Braz. Vogel, K., and Barber, A.A. (1968). For example, HT growth in a medium supplemented with external nitrogen (ammonium) has resulted in the production of cell mass with a protein content of close to 0.7 g/g DW, whereas PT cultivation maximally yields 0.5 g/g DW (Chae et al., 2006; Rodrguez-Zavala et al., 2010). Lipids 47, 913926. Microb. 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). So far, only two E. gracilis enzymes involved in paramylon degradation have been characterised, an endo--1,3-glucanase and a laminaribiose phosphorylase (LBP)/-1,3-glucan phosphorylase (-1,3-GP) involved in paramylon degradation have been characterised (Vogel and Barber, 1968; Takeda et al., 2015; Kuhaudomlarp et al., 2018). PLoS ONE 13, 114. A typical specimen of Euglena gracilis (Fig. Schiff, J.A., Lyman, H., and Russell, G.K. (1980). Lohr, M., Schwender, J., and Polle, J.E.W. 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. Growth and chemical composition of Spirulina maxima and Spirulina platensis biomass at different temperatures. Delta-9 Elongases and Their Use in Making Polyunsaturated Fatty Acids. Cell Res. Nutr. Bull. The effects of dietary restriction and administration of -glucan from Euglena gracilis on the sperm characteristics and reproductive organs of rats. 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. Photosynthesis of Euglena gracilis under cobalamin-sufficient and -limited growing conditions. -1,3-oligoglucan:orthophosphate glucosyltransferases from Euglena gracilis: II. Life Sci. Light dependency of resistance to ionizing radiation in Euglena gracilis. Nutr. 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). Euglena in time: evolution, control of central metabolic processes and multi-domain proteins in carbohydrate and natural product biochemistry. Hepatogastroenterology 56, 547550. Identification and functional analysis of the geranylgeranyl pyrophosphate synthase gene (crtE) and phytoene synthase gene (crtB) for carotenoid biosynthesis in Euglena gracilis. (2018a). doi: 10.1186/s12870-015-0698-8, Keshavarz, B., and Khalesi, M. (2016). 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). J. Vet. 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). For example, intracellular lipids of a mutant strain generated by Fe-ion irradiation of the WT parental strain were stained with a boron-dipyrromethene dye (BODIPY505/515) to isolate high-producers using fluorescence-activated cell sorting. 185186, 922. (2016). Unfortunately, the product yield of this reaction has not been reported (Bumer et al., 2001). Lipids 45, 263273. doi: 10.1007/s00253-008-1443-0, Furuhashi, T., Ogawa, T., Nakai, R., Nakazawa, M., Okazawa, A., Padermschoke, A., et al. 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). of P. gracilis that may compromise the survival of this species but at higher concentrations than recorded in the environment, so P. gracilis can be considered tolerant to this herbicide at environmentally relevant concentrations. A maximum E. gracilis biomass titre of ~48 g DW/L culture was achieved in a fed-batch HT cultivation strictly in the dark, using a jar fermenter (2 L working volume) with glucose concentrations kept constantly above 1 g/L (Ogbonna et al., 1998). However, at present, most studies only extract the paramylon of E. gracilis, which is a waste of resources.In this study, a method of sequential extraction of lipid and paramylon from E. gracilis was studied, and the extraction conditions were optimized. doi: 10.1002/fsn3.828, Nakashima, A., Suzuki, K., Asayama, Y., Konno, M., Saito, K., Yamazaki, N., et al. 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). Prod. Int. Biotechnol. doi: 10.2741/4160, Harper, C. (2006). Immunol. doi: 10.1111/and.13088, Al Abdallah, Q., Nixon, B.T., and Fortwendel, J.R. (2016). doi: 10.1016/j.carbpol.2018.08.005, Shibakami, M., Tsubouchi, G., and Hayashi, M. (2014). The enzymatic conversion of major algal and cyanobacterial carbohydrates to bioethanol. Opt. Immunomodulation and anti-cancer activity of polysaccharide-protein complexes. 142, 121126. Algae need their vitamins. 30, 367373. Ferment. Microbiol. 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). Eur. A more desirable balanced polyunsaturated fatty acid composition achieved by heterologous expression of 15/4 desaturases in mammalian cells. These eukaryotic algae undergo cell division asexually in the dark . Figure 3. doi: 10.1016/S0031-9422(00)90509-0, Ishikawa, T., Masumoto, I., Iwasa, N., Nishikawa, H., Sawa, Y., Shibata, H., et al. Crit. Euglena gracilis paramylon activates human lymphocytes by upregulating pro-inflammatory factors. 4, 115. Vitamin A is found only in animal products (e.g., fish and dairy) and its global human intake is currently supplemented to a large degree with plant-derived provitamin A (-carotene). However, local legal regulations could prevent (outdoor) cultivation of genetically modified E. gracilis strains (Beacham et al., 2017). (2012). Cancer Lett. Paramylon molecules are arranged as an intermolecular triple helix forming microfibrils, which in turn make up fibres. doi: 10.1016/0005-2744(67)90227-6, Matos, .P. J. 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). (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. Bone Mineral Res. doi: 10.1016/0005-2760(92)90311-I, Shigeoka, S., Nakano, Y., and Kitaoka, S. (1979). Biol. Nutr. Lipids 50, 483492. Trends Plant Sci. Commun. Antiviral Res. Purification and properties of laminaribiose phosphorylase (EC 2.4. doi: 10.1007/s11745-012-3690-1, Portnoy, V.A., Bezdan, D., and Zengler, K. (2011). (2018). In the first step of this process, E. gracilis is cultivated under inexpensive PT conditions (Arashida and Mitra, 2015). Euglena live in fresh and brackish water habitats such as ponds rich in organic matter. The Botryococcenes - hydrocarbons of novel structure from the alga Botryococcus braunii, Ktzing. Antioxidants 6, 118. In an alternative approach, genes encoding single enzymes of the E. gracilis PUFA synthesis pathway have been incorporated into PUFA pathways of other organisms to improve the quality of their FA profiles. They possess two flagellae, one long, one short, which can allow the organisms to move. doi: 10.1007/s00253-001-0901-8, Ogbonna, J.C., Tomiyamal, S., and Tanaka, H. (1998). This biomass yield would be comparable to or higher than those obtained with some microalgae grown under HT conditions (Table 1) (Bumbak et al., 2011). Comparison of the production titres of various bioproducts between Euglena gracilis and other source organisms or respective concentrations in alternative products. Assembled and analysed the Euglena gracilis genome and transcriptome in High Performance Computing (HPC) environment with practical experience in Linux environment, and PERL scripting languages. E. gracilis accumulates paramylon during PT, MT, and HT growth (Grimm et al., 2015). doi: 10.1007/s11746-017-3050-7, Maurice, L.Q., Lander, H., Edwards, T., and Harrison, W.E. Miyatake, K., and Kitaoka, S. (1983). Res. 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). Aspects Med. Levine, R.B., LeBrun, J.R., and Horst, G.P. Adv. For example, a fractionation of enzymes secreted by T. harzianum Rifai PAMB-86 led to the enrichment of -1,3-glucanases able to degrade paramylon to some degree, yielding mostly glucose (Giese et al., 2011; Sutivisedsak et al., 2013). Accordingly, alkalised paramylon was shown to upregulate proinflammatory factors (COX-2, IL-6, NO, TNF- and translocation of NF-B) in human lymphomonocytes at a higher rate than the similarly treated commercial -glucan product MacroGuard, which is derived from S. cerevisiae (Russo et al., 2017). For example, Levine et al. doi: 10.1080/00021369.1986.10867518, Shigeoka, S., Toshio, O., Yoshihisa, N., and Shozaburo, K. (1987). Rectangular and wedge-shaped segments consisting of these fibres are arranged to form granules, which can be synthesised by E. gracilis in different shapes like ellipses or rods (Figure 3B). Absorption, transport, and tissue delivery of vitamin E. Mol. Biochim. Euglena gracilis has a plant-like -tocopherol pathway, which has been confirmed by transcriptomic and proteomic studies. Biochem. 23, 12231228. Remarkably, paramylon has been shown to reduce the risk of cancer. Microalgae-based carbohydrates for biofuel production. Determination of the nutritive value of Euglena gracilis protein by in vitro digestion experiments and rat feeding tests. Exp. Vitaminol. Nat. 146, 431442. Comparative studies between laminaribiose- and -1,3-oligoglucan phosphorylase. doi: 10.1023/a:1008011201437, O'Neill, E.C., Trick, M., Henrissat, B., and Field, R.A. (2015a). Carbohydrate Polymers 200, 239247. doi: 10.1007/s11745-015-4010-3, Nandy, S.K., and Srivastava, R.K. (2018). For example, RNAi-mediated silencing of EgWSD isoenzymes resulted in reduced wax ester production in E. gracilis (Teerawanichpan and Qiu, 2010; Tomiyama et al., 2017). Nutr. Omega-3 fatty acid biochemistry: perspectives from human nutrition. Microbiology 133, 2530. Ammonia- and light-induced degradation of paramylum in Euglena gracilis. Many of the last enzymatic steps between the central intermediate isopentenyl pyrophosphate and the end product remain to be elucidated, as only the enzymes geranylgeranyl pyrophosphate synthase and phytoene synthase (see Figure 2A) have been verified by complementing the partially deleted carotenoid biosynthetic pathway of Pantoea ananatis expressed in E. coli with E. gracilis enzymes (Kato et al., 2016). ( 2015a ) doi: 10.1016/0005-2744 ( 67 ) 90227-6, Matos,.P biomass euglena gracilis contributions to the environment bioproduct yields activates lymphocytes! 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