Molecular tools and applications of <i>Euglena gracilis</i>: From biorefineries to bioremediation

Khatiwada, Bishal; Sunna, Anwar; Nevalainen, Helena

doi:10.1002/bit.27516

Cited by 27 publications

(20 citation statements)

References 146 publications

(175 reference statements)

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“…Paramylon is a β-1,3-glucan, that has high biotechnological potential and was considered as a functional food (Nakashima et al, 2018 ; Barsanti and Gualtieri, 2019 ). Several potential biomedical applications of paramylon, such as immunostimulant and an anti-inflammatory agent are under thorough investigations (Nakashima et al, 2017 ; Okouchi et al, 2019 ), and it can serve as a raw material for biofuels, as well (Inui et al, 1982 ; Zimorski et al, 2017 ; Khatiwada et al, 2020 ). The paramylon content decreased upon 200 mM NaCl treatment because it was degraded and converted to soluble sugars, causing trehalose accumulation in the cells (Takenaka et al, 1997 ).…”

Section: Discussionmentioning

confidence: 99%

Salt Stress Induces Paramylon Accumulation and Fine-Tuning of the Macro-Organization of Thylakoid Membranes in Euglena gracilis Cells

et al. 2021

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The effects of salt stress condition on the growth, morphology, photosynthetic performance, and paramylon content were examined in the mixotrophic, unicellular, flagellate Euglena gracilis. We found that salt stress negatively influenced cell growth, accompanied by a decrease in chlorophyll (Chl) content. Circular dichroism (CD) spectroscopy revealed the changes in the macro-organization of pigment-protein complexes due to salt treatment, while the small-angle neutron scattering (SANS) investigations suggested a reduction in the thylakoid stacking, an effect confirmed by the transmission electron microscopy (TEM). At the same time, the analysis of the thylakoid membrane complexes using native-polyacrylamide gel electrophoresis (PAGE) revealed no significant change in the composition of supercomplexes of the photosynthetic apparatus. Salt stress did not substantially affect the photosynthetic activity, as reflected by the fact that Chl fluorescence yield, electron transport rate (ETR), and energy transfer between the photosystems did not change considerably in the salt-grown cells. We have observed notable increases in the carotenoid-to-Chl ratio and the accumulation of paramylon in the salt-treated cells. We propose that the accumulation of storage polysaccharides and changes in the pigment composition and thylakoid membrane organization help the adaptation of E. gracilis cells to salt stress and contribute to the maintenance of cellular processes under stress conditions.

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Section: Discussionmentioning

confidence: 99%

Salt Stress Induces Paramylon Accumulation and Fine-Tuning of the Macro-Organization of Thylakoid Membranes in Euglena gracilis Cells

et al. 2021

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“…Several exogenous materials, such as DNA, RNA, and RNPs (ribonucleoproteins), are the standard delivery objects for bioengineering. An excellent review on the delivery mode of E. gracilis , including protoplast transformation, electroporation, biolistic bombardment, and Agrobacterium -mediated transformation, has been summarized elsewhere and will not be dwelt on here ( Khatiwada et al, 2020 ). It is proposed that single-cell microinjection and electroporation could be the efficient molecule delivery ways for E. gracilis .…”

Section: Harnessing Bioengineering Tools For the Development Of ...mentioning

confidence: 99%

A Synthetic Biology Perspective on the Bioengineering Tools for an Industrial Microalga: Euglena gracilis

Chen

Zhu

et al. 2022

Front. Bioeng. Biotechnol.

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Euglena is a genus of single-celled eukaryotes that show both plant- and animal-like characteristics. Euglena gracilis, a model species, is of great academic interest for studying endosymbiosis and chloroplast development. As an industrial species, E. gracilis is also of primary biotechnological and economic importance as high value-added food, medicine, and cosmetic and high-quality feedstock for jet-fuel production because of its cells containing many high-value products, such as vitamins, amino acids, pigments, unsaturated fatty acids, and carbohydrate paramylon, as metabolites. For more than half a century, E. gracilis has been used as an industrial biotechnology platform for fundamental biology research, mainly exploring relevant physiological and biochemical method studies. Although many researchers focused on genetic engineering tools for E. gracilis in recent years, little progress has been achieved because of the lack of high-quality genome information and efficient techniques for genetic operation. This article reviewed the progress of the genetic transformation of E. gracilis, including methods for the delivery of exogenous materials and other advanced biotechnological tools for E. gracilis, such as CRISPR and RNA interference. We hope to provide a reference to improve the research in functional genomics and synthetic biology of Euglena.

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“…In recent years, there has been an increasing interest in utilizing Euglena as a biofuel producer due to its ability to manufacture waxes suitable for combustion [9,23]. Furthermore, E. gracilis has incited its application as a bioremediation agent [10,[24][25][26] because of its ability to adapt and grow despite various environmental challenges, including low pH and high metal ion concentrations.…”

Section: Introductionmentioning

confidence: 99%

“…Only 20% of the genome assembly or coding sequence ratio has been estimated, supporting the genome's initial features [27,28]. Efficient molecular toolkits and transformation methods are limited for E. gracilis bioengineering compared to other microorganisms of commercial importance [26,29]. Several groups have reported the introduction of double-stranded RNA into Euglena by electroporation and degradation of specific mRNA by the RNAi approach [2,[30][31][32].…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium tumefaciens-Mediated Nuclear Transformation of a Biotechnologically Important Microalga—Euglena gracilis

Becker

Prasad

Ntefidou

et al. 2021

IJMS

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Euglena gracilis (E. gracilis) is an attractive organism due to its evolutionary history and substantial potential to produce biochemicals of commercial importance. This study describes the establishment of an optimized protocol for the genetic transformation of E. gracilis mediated by Agrobacterium (A. tumefaciens). E. gracilis was found to be highly sensitive to hygromycin and zeocin, thus offering a set of resistance marker genes for the selection of transformants. A. tumefaciens-mediated transformation (ATMT) yielded hygromycin-resistant cells. However, hygromycin-resistant cells hosting the gus gene (encoding β-glucuronidase (GUS)) were found to be GUS-negative, indicating that the gus gene had explicitly been silenced. To circumvent transgene silencing, GUS was expressed from the nuclear genome as transcriptional fusions with the hygromycin resistance gene (hptII) (encoding hygromycin phosphotransferase II) with the foot and mouth disease virus (FMDV)-derived 2A self-cleaving sequence placed between the coding sequences. ATMT of Euglena with the hptII-2A–gus gene yielded hygromycin-resistant, GUS-positive cells. The transformation was verified by PCR amplification of the T-DNA region genes, determination of GUS activity, and indirect immunofluorescence assays. Cocultivation factors optimization revealed that a higher number of transformants was obtained when A. tumefaciens LBA4404 (A600 = 1.0) and E. gracilis (A750 = 2.0) cultures were cocultured for 48 h at 19 °C in an organic medium (pH 6.5) containing 50 µM acetosyringone. Transformation efficiency of 8.26 ± 4.9% was achieved under the optimized cocultivation parameters. The molecular toolkits and method presented here can be used to bioengineer E. gracilis for producing high-value products and fundamental studies.

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Molecular tools and applications of Euglena gracilis: From biorefineries to bioremediation

Cited by 27 publications

References 146 publications

Salt Stress Induces Paramylon Accumulation and Fine-Tuning of the Macro-Organization of Thylakoid Membranes in Euglena gracilis Cells

Salt Stress Induces Paramylon Accumulation and Fine-Tuning of the Macro-Organization of Thylakoid Membranes in Euglena gracilis Cells

A Synthetic Biology Perspective on the Bioengineering Tools for an Industrial Microalga: Euglena gracilis

Agrobacterium tumefaciens-Mediated Nuclear Transformation of a Biotechnologically Important Microalga—Euglena gracilis

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