Bio-production of eicosapentaenoic acid from the diatom Nanofrustulum shiloi via two-step high performance countercurrent chromatography

Bárcenas-Pérez, Daniela; Lukeš, Martin; Hrouzek, Pavel; Zápal, Jakub; Kuzma, Marek; Kopeckỳ, J.; Kubáč, David; Arredondo-Vega, Bertha; Cheel, José

doi:10.1007/s10811-022-02816-w

Cited by 5 publications

(5 citation statements)

References 57 publications

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“…In CCC, solvent systems with a wide range of polarities can be used, allowing efficient isolation of compounds with great chemical diversity. Furthermore, since the stationary phase in CCC is liquid, it is possible to inject a large amount of extract and consequently obtain large amounts of fractions/compounds [ 12 , 23 ]. These advantages are particularly important for bioassays and for chemical identification.…”

Section: Discussionmentioning

confidence: 99%

“…Shortly after ME eluted after 30 min, the elution–extrusion mode was applied to the chromatographic process to elute DE, which was strongly retained in the stationary phase. The stationary phase was extruded by switching mobile phase pumping to stationary phase pumping while the column continued to rotate [ 23 ]. The target fractions were collected manually and evaporated at 30 °C under reduced pressure, yielding 1.5 g ME and 0.1 g DE, which were subsequently analyzed using HPLC-DAD.…”

Section: Methodsmentioning

confidence: 99%

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Astaxanthin Extract from Haematococcus pluvialis and Its Fractions of Astaxanthin Mono- and Diesters Obtained by CCC Show Differential Antioxidant and Cytoprotective Effects on Naïve-Mouse Spleen Cells

et al. 2023

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Carotenoids are the most abundant lipid-soluble phytochemicals and are used as dietary supplements to protect against diseases caused by oxidative stress. Astaxanthin, a xanthophyll carotenoid, is a very potent antioxidant with numerous beneficial effects on cellular functions and signaling pathways. In this study, using spleen cells from healthy Balb/c mice, we report the bio-functional effects of an astaxanthin-rich extract (EXT) prepared from the microalga Haematococcus pluvialis and its astaxanthin monoesters-rich fraction (ME) and astaxanthin diesters-rich fraction (DE) obtained by fractionation of EXT using countercurrent chromatography (CCC). After incubation under standard culture conditions (humidity, 37 °C, 5% CO2, atmospheric oxygen), the viability of untreated splenocytes, as determined by the trypan blue exclusion assay, the MTT assay, and the neutral red assay, decreases to approximately 75% after 24 h compared with naïve splenocytes. This effect correlated with the decrease in mitochondrial membrane potential and the transition of ~59% of cells to the early stage of apoptosis, as well as with the decreased ROS production, indicating that hyperoxia in cell-culture deteriorates cell functions. They are restored or stimulated by co-cultivation with EXT, ME, and DE up to 10 µg/mL in the order EXT > DE > ME, suggesting that esterification increases bioavailability to cells in vitro. ROS and H2O2 concentrations reflect mRNA transcriptional activity of Nrf2, superoxide dismutase 1 (SOD1), catalase, and glutathione peroxidase 1, as well as SOD-mediated ROS conversion, whereas they inversely correlate with iNOS-mediated NO production. The highest-tested concentration of EXT, ME, and DE (40 µg/mL) is detrimental to cells, probably because of the overwhelming scavenging activity of astaxanthin and its esters for the reactive oxygen/nitrogen species required for cellular functions and signal transduction at low physiological concentrations. In this study, we demonstrate that differential activities of ME and DE contribute to the final antioxidant and cytoprotective effects of astaxanthin extract, which is beneficial in preventing a wide range of ROS-induced adverse effects, with DE being more effective. In addition, the selection of physioxia-like conditions for pharmacological research is highlighted.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Astaxanthin Extract from Haematococcus pluvialis and Its Fractions of Astaxanthin Mono- and Diesters Obtained by CCC Show Differential Antioxidant and Cytoprotective Effects on Naïve-Mouse Spleen Cells

et al. 2023

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“…Fractional (molecular) distillation utilizes temperature and pressure to manipulate the molecular mean free path and consequently, the evaporation rate of fatty acids with distinct degrees of unsaturation, specifically, to evaporate faster the ones with lower unsaturation (higher molecular mean free path) (Chen et al 2023 ). Chromatographic methods, such as high-performance countercurrent chromatography (Bárcenas-Pérez et al 2022 ), silica-gel chromatography (Sánchez et al 2022 ), or supercritical fluid chromatography (Li et al 2019 ), as well as lipase enzymatic reactions, in which lipase selective hydrolysis can promote the enrichment in EPA (Wang et al 2023 ; da Silva et al 2019 ), have also been applied to concentrate/purify EPA.…”

Section: Downstream Processingmentioning

confidence: 99%

Factors impacting the microbial production of eicosapentaenoic acid

Sousa,

Carvalho,

Gomes

2024

Appl Microbiol Biotechnol

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The increasing applications for eicosapentaenoic acid (EPA) and the potential shortfall in supply due to sustainability and contamination issues related with its conventional sources (i.e., fish oils; seafood) led to an extensive search for alternative and sustainable sources, as well as production processes. The present mini-review covers all the steps involved in the production of EPA from microorganisms, with a deeper focus on microalgae. From production systems to downstream processing, the most important achievements within each area are briefly highlighted. Comparative tables of methodologies are also provided, as well as additional references of recent reviews, so that readers may deepen their knowledge in the different issues addressed. Key points • Microorganisms are more sustainable alternative sources of EPA than fish. • Due to the costly separation from DHA, species that produce only EPA are preferable. • EPA production can be optimised using non-genetic and genetic tailoring engineering.

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“…In another study, eicosapentaenoic acid was isolated by LLC from the biomass of Nanofrustulum shiloi , a species of marine diatoms. The proposed approach led to the high efficiency of the isolation process and high purity of the obtained fatty acid (Bárcenas‐Pérez et al, 2022). At this point, it is important to mention the groundbreaking research of the W. Vetter group, which led to the development of efficient methods for isolating lipid compounds by LLC.…”

Section: Fatty Acids and Plant Oilsmentioning

confidence: 99%

Liquid–liquid chromatography as a promising technology in the separation of food compounds

Kulinowski

Luca

Skalicka‐Woźniak

2023

eFood

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Bio-production of eicosapentaenoic acid from the diatom Nanofrustulum shiloi via two-step high performance countercurrent chromatography

Cited by 5 publications

References 57 publications

Astaxanthin Extract from Haematococcus pluvialis and Its Fractions of Astaxanthin Mono- and Diesters Obtained by CCC Show Differential Antioxidant and Cytoprotective Effects on Naïve-Mouse Spleen Cells

Astaxanthin Extract from Haematococcus pluvialis and Its Fractions of Astaxanthin Mono- and Diesters Obtained by CCC Show Differential Antioxidant and Cytoprotective Effects on Naïve-Mouse Spleen Cells

Factors impacting the microbial production of eicosapentaenoic acid

Liquid–liquid chromatography as a promising technology in the separation of food compounds

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