Overexpression of PtVDL1 in Phaeodactylum tricornutum Increases Fucoxanthin Content under Red Light
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Phaeodactylum tricornutum is a model diatom with significant biotechnological applications, including enhancing biomass, biofuel, and carotenoid production. Specifically, owing to the capacity of this organism to serve as a valuable source of essential raw materials for pharmaceuticals and nutraceuticals, ongoing research is actively focused on enhancing its productivity. One of the genes involved in various stages of fucoxanthin (Fx) biosynthesis, violaxanthin de-epoxidase like 1 (VDL1), has recently been identified. To validate the intracellular function of this gene and boost Fx production through overexpression, we established and examined three transgenic P. tricornutum lines characterized by elevated P. tricortunum VDL1 ( PtVDL1) expression and evaluate their cell growth and Fx productivity. These transgenic lines exhibited substantially increased PtVDL1 mRNA and protein levels compared to the wild type (WT). Notably, the enzyme substrate violaxanthin was entirely depleted and could not be detected in the transformants, whereas it remained at constant levels in the WT. Interestingly, under standard white light conditions, Fx productivity in the transformants remained unchanged; however, but after 48 h of exposure to red light, it increased by up to 15%. These results indicate that PtVDL1-overexpressing P. tricornutum has industrial potential, particularly for enhancing Fx production under red light conditions.
Abstract
Phaeodactylum tricornutum is a model diatom with significant biotechnological applications, including enhancing biomass, biofuel, and carotenoid production. Specifically, owing to the capacity of this organism to serve as a valuable source of essential raw materials for pharmaceuticals and nutraceuticals, ongoing research is actively focused on enhancing its productivity. One of the genes involved in various stages of fucoxanthin (Fx) biosynthesis, violaxanthin de-epoxidase like 1 (VDL1), has recently been identified. To validate the intracellular function of this gene and boost Fx production through overexpression, we established and examined three transgenic P. tricornutum lines characterized by elevated P. tricortunum VDL1 ( PtVDL1) expression and evaluate their cell growth and Fx productivity. These transgenic lines exhibited substantially increased PtVDL1 mRNA and protein levels compared to the wild type (WT). Notably, the enzyme substrate violaxanthin was entirely depleted and could not be detected in the transformants, whereas it remained at constant levels in the WT. Interestingly, under standard white light conditions, Fx productivity in the transformants remained unchanged; however, but after 48 h of exposure to red light, it increased by up to 15%. These results indicate that PtVDL1-overexpressing P. tricornutum has industrial potential, particularly for enhancing Fx production under red light conditions.
참고문헌 (35)
- Armbrust EV, "The life of diatoms in the world’s oceans", Nature, 459, pp.185~192 (2009)
- Field CB, "Primary production of the biosphere : integrating terrestrial and oceanic components", Science, 281, pp.237~240 (1998)
- Aresta M, "Utilisation of CO2 as a chemical feedstock : opportunities and challenges", Dalton Trans, 28, pp.2975~2992 (2007)
- Wijffels RH, "An outlook on microalgal biofuels", Science, 329, pp.796~799 (2010)
- Butler T, "Phaeodactylum tricornutum : a diatom cell factory", Trends Biotechnol, 38, pp.606~622 (2020)
- Murthy KC, "In vivo antioxidant activity of carotenoids from Dunaliella salina-a green microalga", Life Sci, 76, pp.1381~1390 (2005)
- Mulders KJ, "Phototrophic pigment production with microalgae : biological constraints and opportunities", J. Phycol, 50, pp.229~242 (2014)
- Koo SY, "Anti-obesity effect of standardized extract of microalga Phaeodactylum tricornutum containing fucoxanthin", Mar. Drugs, 17, pp.311~ (2019)
- Miyashita K, "Nutraceutical characteristics of the brown seaweed carotenoid fucoxanthin", Arch. Biochem. Biophys, 686, pp.108364~ (2020)
- Pocha CKR, "Current advances in recovery and biorefinery of fucoxanthin from Phaeodactylum tricornutum", Algal Res, 65, pp.102735~ (2022)
- McClure DD, "An investigation into the effect of culture conditions on fucoxanthin production using the marine microalgae Phaeodactylum tricornutum", Algal Res, 29, pp.41~48 (2018)
- Seth K, "Bioprospecting of fucoxanthin from diatoms—challenges and perspectives", Algal Res, 60, pp.102475~ (2021)
- Bai Y, "Green diatom mutants reveal an intricate biosynthetic pathway of fucoxanthin", Proc. Natl. Acad. Sci. U S A, 119, pp.e2203708119~ (2022)
- Bowler C, "The Phaeodactylum genome reveals the evolutionary history of diatom genomes", Nature, 456, pp.239~244 (2008)
- Seo S, "Development of a new constitutive expression system for the transformation of the diatom Phaeodactylum tricornutum", Algal Res, 11, pp.50~54 (2015)
- Seo S, "Enhanced pyruvate metabolism in plastids by overexpression of putative plastidial pyruvate transporter in Phaeodactylum tricornutum", Biotechnol. Biofuels, 13, pp.120~ (2020)
- Bertrand M, "Carotenoid biosynthesis in diatoms", Photosynth. Res, 106, pp.89~102 (2010)
- Dautermann O, "An algal enzyme required for biosynthesis of the most abundant marine carotenoids", Sci. Adv, 6, pp.eaaw9183~ (2020)
- Falciatore A, "Transformation of nonselectable reporter genes in marine diatoms", Mar. Biotechnol, 1, pp.239~251 (1999)
- Baek K, "Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA-free CRISPR-Cas9 RNP-mediated mutagenesis", Biotechnol. Bioeng, 115, pp.719~728 (2018)
- Coesel S, "Evolutionary origins and functions of the carotenoid biosynthetic pathway in marine diatoms", PLoS One, 3, pp.e2896~ (2008)
- Duarte B, "Pigment and fatty acid production under different light qualities in the diatom Phaeodactylum tricornutum", Appl. Sci, 11, pp.2550~ (2021)
- Gupta AK, "Biosynthesis and extraction of high-value carotenoid from algae", Front. Biosci. (Landmark Ed), 26, pp.171~190 (2021)
- Liang MH, "Carotenoids biosynthesis and cleavage related genes from bacteria to plants", Crit. Rev. Food Sci. Nutr, 58, pp.2314~2333 (2018)
- Lohr M, "Algae displaying the diadinoxanthin cycle also possess the violaxanthin cycle", Proc. Natl. Acad. Sci. USA, 96, pp.8784~8789 (1999)
- Kuczynska P, "Photosynthetic pigments in diatoms", Mar. Drugs, 13, pp.5847~5881 (2015)
- Seo S, "Enhanced biomass production by Phaeodactylum tricornutum overexpressing phosphoenolpyruvate carboxylase", Algal Res, 31, pp.489~496 (2018)
- Takaichi S, "Carotenoids in algae : distributions, biosyntheses and functions", Mar. Drugs, 9, pp.1101~1118 (2011)
- Shen X, "A DUF4281 domain-containing protein(homologue of ABA4)of Phaeodactylum tricornutum regulates the biosynthesis of fucoxanthin", Algal Res, 65, pp.102728~ (2022)
- Cao T, "An unexpected hydratase synthesizes the green light-absorbing pigment fucoxanthin", Plant Cell, 35, pp.3053~3072 (2023)
- Zhang H, "Maximizing fucoxanthin production in Odontella aurita by optimizing the ratio of red and blue light-emitting diodes in an auto-controlled internally illuminated photobioreactor", Bioresour. Technol, 344, pp.126260~ (2022)
- Truong TQ, "Interdependence of fucoxanthin biosynthesis and fucoxanthin-chlorophyll a", c binding proteins in Phaeodactylum tricornutum under different light intensities, 2023, pp.35, 25~42
- Yang R, "Improving fucoxanthin production in mixotrophic culture of marine diatom Phaeodactylum tricornutum by LED light shift and nitrogen supplementation", Front. Bioeng. Biotechnol, 8, pp.820~ (2020)
- Schofield A, "Modulation of carotenoid biosynthesis during tomato fruit ripening through phytochrome regulation of phytoene synthase activity", Plant Physiol. Biochem, 43, pp.1052~1060 (2005)
- Fortunato AE, "Diatom phytochromes reveal the existence of farred-light-based sensing in the ocean", Plant Cell, 28, pp.616~628 (2016)