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

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.

[1]  Fu-Li Li,et al.  Rapid Sorting of Fucoxanthin-Producing Phaeodactylum tricornutum Mutants by Flow Cytometry , 2021, Marine drugs.

[2]  R. Wijffels,et al.  Production and high throughput quantification of fucoxanthin and lipids in Tisochrysis lutea using single-cell fluorescence. , 2020, Bioresource technology.

[3]  K. Miyashita,et al.  Nutraceutical characteristics of the brown seaweed carotenoid fucoxanthin. , 2020, Archives of biochemistry and biophysics.

[4]  P. Ralph,et al.  Extrachromosomal genetic engineering of the marine diatom Phaeodactylum tricornutum enables the heterologous production of monoterpenoids. , 2020, ACS synthetic biology.

[5]  Ji-Young Lee,et al.  Health benefits of fucoxanthin in the prevention of chronic diseases. , 2020, Biochimica et biophysica acta. Molecular and cell biology of lipids.

[6]  T. Karpiński,et al.  Fucoxanthin—An Antibacterial Carotenoid , 2019, Antioxidants.

[7]  C. Pan,et al.  Anti-Obesity Effect of Standardized Extract of Microalga Phaeodactylum tricornutum Containing Fucoxanthin , 2019, Marine drugs.

[8]  Ó. Rolfsson,et al.  Chemical Mutagenesis and Fluorescence-Based High-Throughput Screening for Enhanced Accumulation of Carotenoids in a Model Marine Diatom Phaeodactylum tricornutum , 2018, Marine drugs.

[9]  Fu-Li Li,et al.  A Rapid Method for the Determination of Fucoxanthin in Diatom , 2018, Marine drugs.

[10]  Y. Satomi Antitumor and Cancer-preventative Function of Fucoxanthin: A Marine Carotenoid. , 2017, Anticancer research.

[11]  F. Chen,et al.  Screening of Diatom Strains and Characterization of Cyclotella cryptica as A Potential Fucoxanthin Producer , 2016, Marine drugs.

[12]  S. Brynjólfsson,et al.  Photo-Oxidative Stress-Driven Mutagenesis and Adaptive Evolution on the Marine Diatom Phaeodactylum tricornutum for Enhanced Carotenoid Accumulation , 2015, Marine drugs.

[13]  T. Rohrlack,et al.  Spectrophotometric Analysis of Pigments: A Critical Assessment of a High-Throughput Method for Analysis of Algal Pigment Mixtures by Spectral Deconvolution , 2015, PloS one.

[14]  T. Ohama,et al.  Effect of an Introduced Phytoene Synthase Gene Expression on Carotenoid Biosynthesis in the Marine Diatom Phaeodactylum tricornutum , 2015, Marine drugs.

[15]  Wei Li,et al.  Interactive Effects of Ocean Acidification and Nitrogen-Limitation on the Diatom Phaeodactylum tricornutum , 2012, PloS one.

[16]  K. Miyashita,et al.  The allenic carotenoid fucoxanthin, a novel marine nutraceutical from brown seaweeds. , 2011, Journal of the science of food and agriculture.

[17]  A. Falciatore,et al.  Gene silencing in the marine diatom Phaeodactylum tricornutum , 2009, Nucleic acids research.

[18]  N. Bhaskar,et al.  Comparative effects of β-carotene and fucoxanthin on retinol deficiency induced oxidative stress in rats , 2009, Molecular and Cellular Biochemistry.

[19]  Leszek Rychlewski,et al.  The Phaeodactylum genome reveals the evolutionary history of diatom genomes , 2008, Nature.

[20]  Daniel J. Sandberg,et al.  The charge-transfer properties of the S2 state of fucoxanthin in solution and in fucoxanthin chlorophyll-a/c2 protein (FCP) based on stark spectroscopy and molecular-orbital theory. , 2008, The journal of physical chemistry. B.

[21]  R. J. Ritchie,et al.  Universal chlorophyll equations for estimating chlorophylls a, b, c, and d and total chlorophylls in natural assemblages of photosynthetic organisms using acetone, methanol, or ethanol solvents , 2008, Photosynthetica.

[22]  H. Küpper,et al.  Fast, sensitive, and inexpensive alternative to analytical pigment HPLC: quantification of chlorophylls and carotenoids in crude extracts by fitting with Gauss peak spectra. , 2007, Analytical chemistry.

[23]  T. Matsuno Aquatic animal carotenoids , 2001 .

[24]  F. Küpper,et al.  Photometric method for the quantification of chlorophylls and their derivatives in complex mixtures: fitting with Gauss-peak spectra. , 2000, Analytical biochemistry.

[25]  G. Barton,et al.  An investigation into the effect of culture conditions on fucoxanthin production using the marine microalgae Phaeodactylum tricornutum , 2018 .