Column chromatography as a useful step in purification of diatom pigments

  • Wiktor Tokarek Department of Plant Physiology and Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Kraków, Poland
  • Stanisław Listwan Department of Plant Physiology and Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Kraków, Poland
  • Joanna Pagacz Department of Plant Physiology and Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Kraków, Poland
  • Piotr Leśniak Department of Plant Physiology and Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Kraków, Poland
  • Dariusz Latowski Department of Plant Physiology and Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Kraków, Poland
Keywords: column chromatography, Phaeodactylum tricornutum, fucoxanthin, diadinoxanthin, diatoxanthin, thin layer chromatography

Abstract

Fucoxanthin, diadinoxanthin and diatoxanthin are carotenoids found in brown algae and most other heterokonts. These pigments are involved in photosynthetic and photoprotective reactions, and they have many potential health benefits. They can be extracted from diatom Phaeodactylum tricornutum by sonication, extraction with chloroform:methanol and preparative thin layer chromatography. We assessed the utility of an additional column chromatography step in purification of these pigments. This novel addition to the isolation protocol increased the purity of fucoxanthin and allowed the concentration of diadinoxanthin and diatoxanthin before HPLC separation. The enhanced protocol is useful for obtaining high purity pigments for biochemical studies.

References

Aitzetmüller K, Svec WA, Katz JJ, Strain HH (1968) Structure and chemical identity of diadinoxanthin and the principal xanthophyll of Euglena. Chem Commun 1:32-33. http://dx.doi.org/10.1039/C19680000032

Carreto JI, Cataggio JA (1976) Variations in pigment contents of the diatom Phaeodactylum tricornutum during growth. Marine Biology 36:105-112. http://dx.doi.org/10.1007/BF00388433

Choi JH, Kim NH, Kim SJ, Lee HJ, Kim S (2015) Fucoxanthin inhibits the inflammation response in paw edema model through suppressing MAPKs, Akt, and NFκB. J Biochem Mol Toxicol 30:111-119. http://dx.doi.org/10.1002/jbt.21769

Cohen Z (1999) Chemicals from Microalgae. Cohen Z ed, pp 149-160. Taylor & Francis Ltd. ISBN 0-7484-0515-1

Dambek M, Eilers U, Breitenbach J, Steiger S, Büchel C, Sandmann G (2012) Biosynthesis of fucoxanthin and diadinoxanthin and function of initial pathway genes in Phaeodactylum tricornutum. J Exp Bot 63:5607-5612. http://dx.doi.org/10.1093/jxb/ers211

D'Orazio N, Gemello E, Gammone MA, de Girolamo M, Ficoneri C, Riccioni G (2012) Fucoxantin: a treasure from the sea. Mar Drugs 10:604-616. http://dx.doi.org/10.3390/md10030604

Guillard RRL (1975) Culture of phytoplankton for feeding marine invertebrates. In Culture of marine invertebrate animals. Smith WL, Chanley MH eds, pp 26-60. Plenum Press, New York, USA. http://dx.doi.org/10.1007/978-1-4615-8714-9_3

Haugan JA, Liaaen-Jensen S (1989) Improved isolation procedure for fucoxanthin. Phytochemistry 28:2797-2798. http://dx.doi.org/10.1016/S0031-9422(00)98091-9

Jeffrey SW, Mantoura RFC, Bjørnland T (1997) Data for identification of 47 key phytoplankton pigments. In Phytoplankton pigments in oceanography: guidelines to modern methods. Jeffrey SW, Mantoura RFC, Wright SW eds, pp 493-560. UNESCO Publishing.

Johansen JE, Svec WA, Liaaen-Jensen S, Haxo FT (1974) Carotenoids of the Dinophyceae. Phytochemistry 13:2261-2271. http://dx.doi.org/10.1016/0031-9422(74)85038-7

Johnsen G, Lysaa PA, Aamodt K (2014) Sunscreen Compositions Comprising Carotenoids. Promar AS, assignee. Patent US 8834855 B2.

Konishi I, Hosokawa M, Sashima T, Maoka T, Miyashita K (2008) Suppressive effects of alloxanthin and diatoxanthin from Halocynthia roretzi on LPS-induced expression of pro-inflammatory genes in RAW264.7 cells. J Oleo Sci 57:181-189. http://dx.doi.org/10.5650/jos.57.181

Kraay GW, Zapata M, Veldhuis MJW (1992) Separation of chlorophylls c1, c2, and c3 of marine phytoplankton by reversed-phase-C18-high-performance liquid chromatography. J Phycol 28:708-712.

http://dx.doi.org/10.1111/j.0022-3646.1992.00708.x

Krieger-Liszkay A (2005) Singlet oxygen production in photosynthesis. J Exp Bot 56:337-346. http://dx.doi.org/10.1093/jxb/erh237

Kumar SR, Hosokawa M, Miyashita K (2013) Fucoxanthin: A Marine Carotenoid Exerting Anti-Cancer Effects by Affecting Multiple Mechanisms. Mar Drugs 11:5130–514. http://dx.doi.org/10.3390/md11125130

Latowski D, Kuczyńska P, Strzałka K (2011) Xanthophyll cycle - a mechanism protecting plants against oxidative stress. Redox Report 16:78-90. http://dx.doi.org/10.1179/174329211X13020951739938

Lavaud J, Rousseau B, van Gorkom HJ, Etienne AL (2002) Influence of the diadinoxanthin pool size on photoprotection in the marine planktonic diatom Phaeodactylum tricornutum Plant Physiol 129:1398-1406. http://dx.doi.org/10.1104/pp.002014

Liaaen-Jensen S (1989) Artifacts of natural carotenoids - unintended carotenoid synthesis. In Carotenoids: chemistry and biology. Krinsky NI, Mathews-Roth MM, Taylor RF eds, pp 149-165. Plenum Press, New York, USA. http://dx.doi.org/10.1007/978-1-4613-0849-2_9

Marder JB, Droppa M, Casp V, Raskin VI, Horváth G (1998) Light-independent thermoluminescence from thylakoids of greening barley leaves. Evidence for involvement of oxygen radicals and free chlorophyll. Physiologia Plantarum 104:713-719. http://dx.doi.org/10.1034/j.1399-3054.1998.1040428.x

Mikami K, Hosokawa M (2013) Biosynthetic pathway and health benefits of fucoxanthin, an algae-specific xanthophyll in brown seaweeds. Int J Mol Sci 14:13763–13781. http://dx.doi.org/10.3390/ijms140713763

Miyashita K, Hosokawa M (2015) The beneficial health effects of fucoxanthin. In Genomics, proteomics and metabolomics in nutraceuticals and functional foods, second edition. Bagchi D, Swaroop A, Bagchi M eds, pp 122-134. John Wiley & Sons, Ltd, Chichester, UK. http://dx.doi.org/10.1002/9781118930458.ch10

Moghadamtousi SZ, Karimian H, Khanabdali R, Razavi M, Firoozinia M, Zandi K, Kadir HA (2014) Anticancer and antitumor potential of fucoidan and fucoxanthin, two main metabolites isolated from brown algae. The Scientific World Journal, Article ID 768323. http://dx.doi.org/10.1155/2014/768323

Nomura T, Kikuchi M, Kubodera A, Kawakami Y (1997) Proton-donative antioxidant activity of fucoxanthin with 1,1-diphenyl-2-picrylhydrazyl (DPPH). Biochem Mol Biol Int 42:361-70. http://dx.doi.org/10.1080/15216549700202761

Papagiannakis E, van Stokkum IHM, Fey H, Büchel C, van Grondelle R (2005) Spectroscopic characterization of the excitation energy transfer in the fucoxanthin–chlorophyll protein of diatoms. Photosynth Res 86:241-250. http://dx.doi.org/10.1007/s11120-005-1003-8

Pfeil BE, Schoefs B, Spetea C (2014) Function and evolution of channels and transporters in photosynthetic membranes. Cell Mol Life Sci 71:979-998. http://dx.doi.org/ 10.1007/s00018-013-1412-3

R Core Team (2016) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL https://www.R-project.org/.

Ritchie RJ (2006) Consistent sets of spectrophotometric chlorophyll equations for acetone, methanol and ethanol solvents. Photosynth Res 89:27-41. http://dx.doi.org/10.1007/s11120-006-9065-9

Sachindra NM, Sato E, Maeda H, Hosokawa M, Niwano Y, Kohno M, Miyashita K (2007) Radical scavenging and singlet oxygen quenching activity of marine carotenoid fucoxanthin and its metabolites. J Agric Food Chem 55:8516-8522. http://dx.doi.org/10.1021/jf071848a

Sadura I (2014) Isolation and purification of diadinoxanthin cycle pigments and their influence on molecular dynamic of artificial membranes (in Polish). Master’s thesis, Faculty of Chemistry, Jagiellonian University in Kraków, Poland.

Published
2016-08-02
Section
Articles