Production and high throughput quantification of fucoxanthin and lipids in Tisochrysis lutea using single-cell fluorescence

Gao, Fengzheng; Teles, Iago; Ferrer-Ledo, Narcís; Wijffels, René H.; Barbosa, María J.

doi:10.1016/j.biortech.2020.124104

Cited by 30 publications

(16 citation statements)

References 45 publications

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“…Total lipids (24, 31 and 21% of algal DW, respectively for mono-cultures of N. oculata and T. lutea, and for the co-cultures) did not differ significantly between mono-and cocultivated algae (p > 0.2; one-way ANOVA). The results for mono-cultures were substantially in line with those available in the literature for the same algae (Su et al 2011;Gao et al 2020). Interestingly, the lower, even if not significant, value obtained for the co-cultures reflected the different proportion of algae in the whole population, where N. oculata was dominant over T. lutea.…”

Section: Lipid and Fatty Acid Profilesupporting

confidence: 89%

A co-cultivation process of Nannochloropsis oculata and Tisochrysis lutea induces morpho-physiological and biochemical variations potentially useful for biotechnological purposes

et al. 2021

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The biotechnological potential of microalgae has gained considerable importance in many applied fields: biomass production for food and feed, cosmeceutical and pharmaceutical products, energy and phytoremediation. The driving force that inspires the progress in microalgae production is the need for new cultivation systems to obtain simultaneously the maximum yield, reduction of water and nutrients use, and production of economically interesting molecules, such as pigments, fatty acids and polysaccharides. We aim to test, for the first time, the co-cultivation in saline medium of Tisochrysis lutea (Haptophyta) and Nannochloropsis oculata (Ochrophyta) to obtain valuable compounds, i.e. pigments and lipids characteristic of each species, using a single culture process. Mono-cultures of each strain were used as controls. The two strains showed an increase in the concentration of chlorophylls and carotenoids in co-culture. At the end of the experiment, the fatty acid profile was analysed by gas chromatography–mass spectrometry. The lipids in the co-cultivated cell extracts were mainly attributable to N. oculata, which represented 97% of the total cells (ca. 83% of the total biomass) at the end of the experiment. Nevertheless, the ω-3 characteristic of T. lutea (DHA and SDA, absent in N. oculata) was also detectable. Although the co-cultivation of these two phylogenetically different species of microalgae did not show positive effects on the growth and on the total lipid production, however, this process resulted in a reduction of the production costs and a lower consumption of water and nutrients.

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Section: Lipid and Fatty Acid Profilesupporting

confidence: 89%

A co-cultivation process of Nannochloropsis oculata and Tisochrysis lutea induces morpho-physiological and biochemical variations potentially useful for biotechnological purposes

et al. 2021

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“…The use of Nile Red to correlate polar lipids with fucoxanthin has been performed previously using live cultures of Tisochrysis lutea with an R 2 value of 0.88 [17], there is a correlation between Nile Red and total carotenoids in P. tricornutum [15]. A relationship to HPLC was found herein (R 2 = 0.69) for P. tricornutum using the average single cell fluorescence of singlets after staining with Nile Red (Figure 2c).…”

Section: High-throughput Screen Analysissupporting

confidence: 54%

“…Measuring autofluorescence with flow cytometry is not only accurate for measuring fucoxanthin, it does not require extraction, dilution or dyeing, and this greatly improves screening time. Relevant to many projects that generate large mutant or transformant libraries, FACS can also be attached to this method to further improve screening and selection times, as also found in Gao et al [17] and Fan et al [25].…”

Section: High-throughput Screen Analysismentioning

confidence: 99%

“…First, two estimates for chlorophyll and fucoxanthin autofluorescence were measured using flow cytometry to investigate whether various excitation/emission gates could accurately predict average fucoxanthin content through analysis of single cells. A third flow cytometry method using Nile Red dye was included to investigate if dye fluorescence can improve the detection of fucoxanthin based on its relationship to lipids [17] and correlation to total carotenoids [15]. One approach for estimating chlorophyll in ethanol was tested as a proxy for measuring fucoxanthin in microplate format from Ritchie [18].…”

Section: Introductionmentioning

confidence: 99%

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Comparative Study Highlights the Potential of Spectral Deconvolution for Fucoxanthin Screening in Live Phaeodactylum tricornutum Cultures

et al. 2021

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Microalgal biotechnology shows considerable promise as a sustainable contributor to a broad range of industrial avenues. The field is however limited by processing methods that have commonly hindered the progress of high throughput screening, and consequently development of improved microalgal strains. We tested various microplate reader and flow cytometer methods for monitoring the commercially relevant pigment fucoxanthin in the marine diatom Phaeodactylum tricornutum. Based on accuracy and flexibility, we chose one described previously to adapt to live culture samples using a microplate reader and achieved a high correlation to HPLC (R2 = 0.849), effectively removing the need for solvent extraction. This was achieved by using new absorbance spectra inputs, reducing the detectable pigment library and changing pathlength values for the spectral deconvolution method in microplate reader format. Adaptation to 384-well microplates and removal of the need to equalize cultures by density further increased the screening rate. This work is of primary interest to projects requiring detection of biological pigments, and could theoretically be extended to other organisms and pigments of interest, improving the viability of microalgae biotechnology as a contributor to sustainable industry.

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“…The OD was measured using a DR 5000™ UV‐Vis Laboratory Spectrophotometer (Hach) in duplicate. DW was measured according to a method described previously (Gao et al, 2020a).…”

Section: Methodsmentioning

confidence: 99%

Production and monitoring of biomass and fucoxanthin with brown microalgae under outdoor conditions

Gao

Sá

Itd

et al. 2020

Biotech & Bioengineering

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The effect of light on biomass and fucoxanthin (Fx) productivities was studied in two microalgae, Tisochrysis lutea and Phaeodactylum tricornutum. High and low biomass concentrations (1.1 and 0.4 g L−1) were tested in outdoor pilot‐scale flat‐panel photobioreactors at semi‐continuous cultivation mode. Fluorescence spectroscopy coupled with chemometric modeling was used to develop prediction models for Fx content and for biomass concentration to be applied for both microalgae species. Prediction models showed high R2 for cell concentration (.93) and Fx content (.77). Biomass productivity was lower for high biomass concentration than low biomass concentration, for both microalgae (1.1 g L−1: 75.66 and 98.14 mg L−1 d−1, for T. lutea and P. tricornutum, respectively; 0.4 g L−1: 129.9 and 158.47 mg L−1 d−1, T. lutea and P. tricornutum). The same trend was observed in Fx productivity (1.1 g L−1: 1.14 and 1.41 mg L−1 d−1, T. lutea and P. tricornutum; 0.4 g L−1: 2.09 and 1.73 mg L−1 d−1, T. lutea and P. tricornutum). These results show that biomass and Fx productivities can be set by controlling biomass concentration under outdoor conditions and can be predicted using fluorescence spectroscopy. This monitoring tool opens new possibilities for online process control and optimization.

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Production and high throughput quantification of fucoxanthin and lipids in Tisochrysis lutea using single-cell fluorescence

Cited by 30 publications

References 45 publications

A co-cultivation process of Nannochloropsis oculata and Tisochrysis lutea induces morpho-physiological and biochemical variations potentially useful for biotechnological purposes

A co-cultivation process of Nannochloropsis oculata and Tisochrysis lutea induces morpho-physiological and biochemical variations potentially useful for biotechnological purposes

Comparative Study Highlights the Potential of Spectral Deconvolution for Fucoxanthin Screening in Live Phaeodactylum tricornutum Cultures

Production and monitoring of biomass and fucoxanthin with brown microalgae under outdoor conditions

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