Selection and characterization of Euglena anabaena var. minor as a new candidate Euglena species for industrial application

Suzuki, Kengo; Mitra, Sharbanee; Iwata, Osamu; Ishikawa, Takahiro; Kato, Sueo; Yamada, Koji

doi:10.1080/09168451.2015.1045828

Cited by 23 publications

(21 citation statements)

References 15 publications

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“…Indeed, a species, Euglena (Euglenaria) anabaena var. minor, has been found to proliferate relatively faster than the other Euglena species, and has the preferable characteristic of rapid sinking, which will facilitate its harvesting process (Suzuki et al 2015).…”

Section: Breeding Potential Of Euglenamentioning

confidence: 99%

Large-Scale Cultivation of Euglena

Suzuki¹

2017

Advances in Experimental Medicine and Biology

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From the middle of the twentieth century, microalgae have been exploited as a candidate biomass source of food and other products. One such candidate source is the fast-proliferating microalga Euglena gracilis. The commercial cultivation of E. gracilis began in 2007, after the success of its outdoor mass cultivation and improvement of the harvesting and drying methods suitable for Euglena cells. The commercialization of Euglena production is based on the strategy of "5Fs of Biomass," which refers to the development and production of commercial products including food, fiber, feed, fertilizer, and fuel from biomass." Although room for improvement remains in the productivity of Euglena biomass, the product with the highest value-food-is already profitable. By enhancing the productivity of its biomass, other Euglena products, including fiber, feed, fertilizer, and fuel, can be commercialized. Breeding and recombinant DNA technology studies are being conducted to accomplish more extensive application of Euglena. In addition, the search for a better place for outdoor mass cultivation of Euglena is ongoing.

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Section: Breeding Potential Of Euglenamentioning

confidence: 99%

Large-Scale Cultivation of Euglena

Suzuki¹

2017

Advances in Experimental Medicine and Biology

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“…For example, E. anabaena var. minor precipitates easily 37 , E. mutabilis grows in acid mine drainages 38 , and E. sanguinea produces carotenoid astaxantin 39 . While not included in this Article, we have attempted and verified that FACS analysis can also be applied to E. anabaena var.…”

Section: Discussionmentioning

confidence: 99%

“…Approximately 10 mg and 100 mg of samples of dried cells were used for quantifying their intracellular paramylon and lipid content, respectively. The paramylon content was evaluated in the same way as in previous reports 13 37 . The dried cells were briefly suspended in 10 mL of acetone and homogenized twice for 90 sec by using a sonicator (UD-201, TOMY).…”

Section: Methodsmentioning

confidence: 99%

Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting

Yamada¹,

Suzuki²,

Takeuchi³

et al. 2016

Sci Rep

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Euglena gracilis, a microalgal species of unicellular flagellate protists, has attracted much attention in both the industrial and academic sectors due to recent advances in the mass cultivation of E. gracilis that have enabled the cost-effective production of nutritional food and cosmetic commodities. In addition, it is known to produce paramylon (β-1,3-glucan in a crystalline form) as reserve polysaccharide and convert it to wax ester in hypoxic and anaerobic conditions–a promising feedstock for biodiesel and aviation biofuel. However, there remain a number of technical challenges to be solved before it can be deployed in the competitive fuel market. Here we present a method for efficient selective breeding of live oil-rich E. gracilis with fluorescence-activated cell sorting (FACS). Specifically, the selective breeding method is a repetitive procedure for one-week heterotrophic cultivation, staining intracellular lipids with BODIPY505/515, and FACS-based isolation of top 0.5% lipid-rich E. gracilis cells with high viability, after inducing mutation with Fe-ion irradiation to the wild type (WT). Consequently, we acquire a live, stable, lipid-rich E. gracilis mutant strain, named B1ZFeL, with 40% more lipid content on average than the WT. Our method paves the way for rapid, cost-effective, energy-efficient production of biofuel.

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“…E. gracilis cells were dried in a freeze dryer (Lyph-Lock 6, Labconco) after counting the cells. Paramylon in the dried cells was extracted and quantified by using the phenol-sulfuric acid method 44 , 45 . The average amount of intracellular paramylon per cell was estimated by dividing the total extracted amount by the cell population.…”

Section: Methodsmentioning

confidence: 99%

Targeted delivery of fluorogenic peptide aptamers into live microalgae by femtosecond laser photoporation at single-cell resolution

Maeno

Uzawa

Kono

et al. 2018

Sci Rep

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Microalgae-based metabolic engineering has been proven effective for producing valuable substances such as food supplements, pharmaceutical drugs, biodegradable plastics, and biofuels in the past decade. The ability to accurately visualize and quantify intracellular metabolites in live microalgae is essential for efficient metabolic engineering, but remains a major challenge due to the lack of characterization methods. Here we demonstrate it by synthesizing fluorogenic peptide aptamers with specific binding affinity to a target metabolite and delivering them into live microalgae by femtosecond laser photoporation at single-cell resolution. As a proof-of-principle demonstration of our method, we use it to characterize Euglena gracilis, a photosynthetic unicellular motile microalgal species, which is capable of producing paramylon (a carbohydrate granule similar to starch). Specifically, we synthesize a peptide aptamer containing a paramylon-binding fluorescent probe, 7-nitrobenzofurazan, and introduce it into E. gracilis cells one-by-one by suppressing their mobility with mannitol and transiently perforating them with femtosecond laser pulses at 800 nm for photoporation. To demonstrate the method’s practical utility in metabolic engineering, we perform spatially and temporally resolved fluorescence microscopy of single live photoporated E. gracilis cells under different culture conditions. Our method holds great promise for highly efficient microalgae-based metabolic engineering.

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Selection and characterization of Euglena anabaena var. minor as a new candidate Euglena species for industrial application

Cited by 23 publications

References 15 publications

Large-Scale Cultivation of Euglena

Large-Scale Cultivation of Euglena

Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting

Targeted delivery of fluorogenic peptide aptamers into live microalgae by femtosecond laser photoporation at single-cell resolution

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