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

Marine Drugs ◽  
2021 ◽  
Vol 20 (1) ◽  
pp. 19
Author(s):  
Sean Macdonald Miller ◽  
Raffaela M. Abbriano ◽  
Anna Segecova ◽  
Andrei Herdean ◽  
Peter J. Ralph ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Phaeodactylum Tricornutum ◽  
Marine Diatom ◽  
Screening Rate ◽  
Primary Interest ◽  
Spectral Deconvolution ◽  
Microalgal Biotechnology ◽  
Microplate Reader ◽  
Spectral Deconvolution Method ◽  
Live Culture

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.

scholarly journals Fluorescent Artificial Receptor-Based Membrane Assay (FARMA) for Spatiotemporally Resolved Monitoring of Biomembrane Permeability

2019 ◽  
Author(s):  
Frank Biedermann ◽  
Garima Ghale, Ghale ◽  
Andreas Hennig ◽  
Werner M. Nau
Keyword(s):  
High Throughput Screening ◽  
High Sensitivity ◽  
Point Of View ◽  
Phospholipid Bilayer ◽  
Label Free ◽  
Microscopy Imaging ◽  
Biophysical Models ◽  
Fundamental Point ◽  
Artificial Receptor ◽  
Microplate Reader

<p>The spatiotemporally resolved monitoring of membrane translocation, <i>e.g.,</i> of drugs or toxins, has been a long-standing goal. Herein, we introduce the fluorescent artificial receptor-based membrane assay (FARMA), a facile, label-free method. With FARMA, the permeation of more than hundred organic compounds (drugs, toxins, pesticides, neurotransmitters, peptides, etc.) through vesicular phospholipid bilayer membranes has been monitored in real time (µs-h time scale) and with high sensitivity (nM-µM concentration), affording permeability coefficients across an exceptionally large range from 10<sup>–9</sup>‑10<sup>–3</sup> cm s<sup>–1</sup>. From a fundamental point of view, FARMA constitutes a powerful tool to assess structure-permeability relationships and to test biophysical models for membrane passage. From an applied perspective, FARMA can be extended to high-throughput screening by adaption of the microplate reader format, to spatial monitoring of membrane permeation by microscopy imaging, and to the compartmentalized monitoring of enzymatic activity.<b></b></p>

scholarly journals Chemical Mutagenesis and Fluorescence-Based High-Throughput Screening for Enhanced Accumulation of Carotenoids in a Model Marine Diatom Phaeodactylum tricornutum

Marine Drugs ◽  
2018 ◽  
Vol 16 (8) ◽  
pp. 272 ◽  
Author(s):  
Zhiqian Yi ◽  
Yixi Su ◽  
Maonian Xu ◽  
Andreas Bergmann ◽  
Saevar Ingthorsson ◽  
...  
Keyword(s):  
High Throughput ◽  
Metabolic Pathways ◽  
Phaeodactylum Tricornutum ◽  
Mass Spectrometry Data ◽  
Marine Diatom ◽  
Chemical Mutagenesis ◽  
Carotenoid Content ◽  
Enzymatic Reactions ◽  
Wild Type ◽  
A Genome

Diatoms are a major group of unicellular algae that are rich in lipids and carotenoids. However, sustained research efforts are needed to improve the strain performance for high product yields towards commercialization. In this study, we generated a number of mutants of the model diatom Phaeodactylum tricornutum, a cosmopolitan species that has also been found in Nordic region, using the chemical mutagens ethyl methanesulfonate (EMS) and N-methyl-N′-nitro-N-nitrosoguanidine (NTG). We found that both chlorophyll a and neutral lipids had a significant correlation with carotenoid content and these correlations were better during exponential growth than in the stationary growth phase. Then, we studied P. tricornutum common metabolic pathways and analyzed correlated enzymatic reactions between fucoxanthin synthesis and pigmentation or lipid metabolism through a genome-scale metabolic model. The integration of the computational results with liquid chromatography-mass spectrometry data revealed key compounds underlying the correlative metabolic pathways. Approximately 1000 strains were screened using fluorescence-based high-throughput method and five mutants selected had 33% or higher total carotenoids than the wild type, in which four strains remained stable in the long term and the top mutant exhibited an increase of 69.3% in fucoxanthin content compared to the wild type. The platform described in this study may be applied to the screening of other high performing diatom strains for industrial applications.

Structural basis for blue-green light harvesting and energy dissipation in diatoms

Science ◽  
2019 ◽  
Vol 363 (6427) ◽  
pp. eaav0365 ◽  
Author(s):  
Wenda Wang ◽  
Long-Jiang Yu ◽  
Caizhe Xu ◽  
Takashi Tomizaki ◽  
Songhao Zhao ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Phaeodactylum Tricornutum ◽  
Light Harvesting ◽  
Green Light ◽  
Marine Diatom ◽  
Structural Basis ◽  
Aquatic Environments ◽  
Efficient Energy Transfer ◽  
Chl A ◽  
Photosynthetic Organisms

Diatoms are abundant photosynthetic organisms in aquatic environments and contribute 40% of its primary productivity. An important factor that contributes to the success of diatoms is their fucoxanthin chlorophyll a/c-binding proteins (FCPs), which have exceptional light-harvesting and photoprotection capabilities. Here, we report the crystal structure of an FCP from the marine diatom Phaeodactylum tricornutum, which reveals the binding of seven chlorophylls (Chls) a, two Chls c, seven fucoxanthins (Fxs), and probably one diadinoxanthin within the protein scaffold. Efficient energy transfer pathways can be found between Chl a and c, and each Fx is surrounded by Chls, enabling the energy transfer and quenching via Fx highly efficient. The structure provides a basis for elucidating the mechanisms of blue-green light harvesting, energy transfer, and dissipation in diatoms.

The roles of silicon in combating cadmium challenge in the Marine diatom Phaeodactylum tricornutum

2020 ◽  
Vol 389 ◽  
pp. 121903 ◽  
Author(s):  
Jie Ma ◽  
Beibei Zhou ◽  
Qiaoguo Tan ◽  
Li Zhang ◽  
Ke Pan
Keyword(s):  
Phaeodactylum Tricornutum ◽  
Marine Diatom
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