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High-performance countercurrent chromatography for lutein production from a chlorophyll-deficient strain of the microalgae Parachlorella kessleri HY1

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Abstract

Green microalgae are a recognized lutein source; however, processing for lutein production requires additional operations such as extraction and saponification, mainly due to the high green pigment and lipid content in the biomass. In this study lutein was isolated from a chlorophyll-deficient Parachlorella kessleri HY1 strain using high-performance countercurrent chromatography (HPCCC). The lower phase of the biphasic solvent system composed of n-heptane–ethanol–water, 5:4:1.5, v/v/v was used both as biomass extraction solvent and HPCCC mobile phase conferring a high selectivity to the lutein production process. For the HPCCC isolation, a multiple injection method was developed, and ten consecutive sample injections (300 mg per each) were performed. To favor the economics of the process, the HPCCC mobile and stationary phases were separately formulated based on nuclear magnetic resonance (NMR) analyses. This strategy enabled to avoid obtaining immiscible liquid phases from their parent biphasic solvent system, which led to the reduction of the separation process duration and solvent consumption. Overall, 3 g of P. kessleri HY1 strain extract was processed by HPCCC yielding 150 mg of lutein (95% purity, 97% recovery). The results presented here form an efficient and economical basis for the large-scale production of microalgae-sourced lutein.

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Acknowledgements

This work was financially supported by the MPO-TRIO grant (reg. č. FV10155, JK, JC, DK), and the National Programme of Sustainability I of the Ministry of Education, Youth and Sports of the Czech Republic (ID: LO1416, JC, TF, PH, JK, DK). T. Fábryová gratefully acknowledges the scientific supervision of Dr. José Cheel (Centre Algatech–Czech Academy of Sciences) during the PhD study, as well as the support of the SVV grant (260416) and the Charles University for a PhD scholarship.

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Authors and Affiliations

  1. Laboratory of Algal Biotechnology–Centre ALGATECH, Institute of Microbiology of the Czech Academy of Sciences, Opatovický mlýn, 379 81, Třeboň, Czech Republic

    Tereza Fábryová, David Kubáč, Pavel Hrouzek, Jiří Kopecký & José Cheel

  2. Department of Pharmacognosy, Faculty of Pharmacy, Charles University, Akademika Heyrovského 1203, 500 05, Hradec Králové, Czech Republic

    Tereza Fábryová & Lenka Tůmová

  3. Laboratory of Molecular Structure Characterization, Institute of Microbiology of the Czech Academy of Sciences, Vídeňská 1083, 142 20, Prague, Czech Republic

    Marek Kuzma

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  1. Tereza Fábryová
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Contributions

José Cheel conceived and designed the HPCCC experiments and wrote the article. Tereza Fábryová and José Cheel performed the extraction and isolation experiments, analyzed the data, and revised the manuscript. David Kubáč performed the microalgae cultivation, analyzed data, and revised the manuscript. Jiří Kopecký and Pavel Hrouzek performed qualitative MS analysis of the samples and revised the manuscript. Marek Kuzma performed NMR analysis and revised the manuscript. Lenka Tůmová assisted in the data analysis and revised the manuscript.

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Correspondence to José Cheel.

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Fábryová, T., Kubáč, D., Kuzma, M. et al. High-performance countercurrent chromatography for lutein production from a chlorophyll-deficient strain of the microalgae Parachlorella kessleri HY1. J Appl Phycol 33, 1999–2013 (2021). https://doi.org/10.1007/s10811-021-02434-y

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