Supercritical Fluid Extraction of Lutein Esters from Marigold Flowers and their Hydrolysis by Improved Saponification and Enzyme Biocatalysis

Authors: A. Peter Amala Sujith, T.V. Hymavathi, P. Yasoda Devi

Abstract:

Lutein is a dietary oxycarotenoid which is found to reduce the risks of Age-related Macular Degeneration (AMD). Supercritical fluid extraction of lutein esters from marigold petals was carried out and was found to be much effective than conventional solvent extraction. The saponification of pre-concentrated lutein esters to produce free lutein was studied which showed a composition of about 88% total carotenoids (UV-VIS spectrophotometry) and 90.7% lutein (HPLC). The lipase catalyzed hydrolysis of lutein esters in conventional medium was investigated. The optimal temperature, pH, enzyme concentration and water activity were found to be 50°C, 7, 15% and 0.33 respectively and the activity loss of lipase was about 25% after 8 times re-use in at 50°C for 12 days. However, the lipase catalyzed hydrolysis of lutein esters in conventional media resulted in poor conversions (16.4%).

Keywords: lutein, preconcentration, saponification, lipase

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1331475

References:

[1] D.E Breithaupt, A. Bamedi and U. Wirt,. Carotenol fatty acid esters: Easy substrates for digestive enzymes? Comp Biochem Physiol B Biochem Mol Biol. 2002, 132: pp. 721-728.
[2] Bunea Andrea.. Lutein esters from Tagetes erecta: Isolation and enzymatic hydrolysis. Bulletin UASVM .Animal Science and Biotechnologies. 2008, 65:pp.1-2.
[3] L. Chasan-Taber, W.C. Willet, J.M. Seddon, M.J. Stampfer, B.Rosner, G.A. Colditz, F.E. Speizer and S.E.Hankinson. A prospective study of carotenoid and vitamin A intakes and risk of cataract extraction in US women. Am J Clin Nutr. 1999.70: pp.509-16.
[4] H.K. Chung, H.M. Rasmussen and E.J. Johnson. Lutein bioavailability is higher from lutein-enriched eggs than from supplements and spinach in men. J. Nutr. 2004.134(8):pp.1887-1893.
[5] R.E. Feeney. Chemical deterioration of proteins. Edited by J.R. Whitaker and M. Fujimaki. American Chemical Society. Washington. D.C.1980.pp.1-99.
[6] H. L. Goderis, B. L. Fouwe, S. M. Cauwenbergh and P. P. Tobback. Measurement and control of water content of organic solvents. Anal. Chem. 1986.58: pp.1561-1563.
[7] G. Gore. On the properties of liquid carbonic acid. Philosophical Transactions of the Royal Society of London. Series A: Mathematical, Physical and Engineering Sciences. 1861.pp.151:83.
[8] F. Granado, B. Olmedilla and I. Blanco. Serum depletion and bioavailabilty of lutein in type I diabetic patients. Eur J Nutr. 2002.41(2):pp.47-53.
[9] J. B. Hannay and J. Hogarth. On the solubility of solids in gases. Proceedings of the Royal Society of London Series A. 1879.pp.29:324.
[10] F.C. Huang, Y.H. Ju and C.W. Huang. Enrichment of ╬│- linolenic acid from borage oil via lipase-catalyzed reactions. J. Am. Oil Chem. Soc. 1997. 74: pp.977-981.
[11] M. Ikeda. Public health problems of organic solvents. Toxicol Lett. 1992. 64-65: pp.191-201.
[12] P.B. Jacobs, R.D. Le Boeuf, S.A. McCommas and J.D. Tauber. The cleavage of carotenoid esters by cholesterol esterase. ? Comp Biochem Physiol B Biochem Mol Biol. 1982. 72B: 157-160.
[13] A.J. Jay, D.C. Steytler and M. Knights. Spectrophotometric studies of food colors in near critical CO2. J Supercrit Fluids. 1991.4: pp.131-141.
[14] J.A. Kayanush, T.B.Hannah, M. Paula and M. Barry. Lutein is stable in yogurt and does not affect its charecteristics. J Food Sci. 2006.71(6): pp.467-472.
[15] F. Khachik. Process for isolation, purification and recrystallization of lutein from marigold oleoresin and uses thereof. United States Patent 5382714. 1995.
[16] T.K. Kumar. Trans-lutein enriched xanthophylls ester concentrate and a process for its preparation. 2004. United States Patent 6737535.
[17] J.T Landrum and R.A Bone. Lutein, zeaxanthin and macular pigment. Arch Biochem Biophys. 2001.385:pp.28-40.
[18] E.W. Lusas and S. R. Gregory. New solvents and extractors. In: Proceedings of the World Conference on Oilseed and Edible Oils Processing.. Koseoglu, S.S., Rhee, K.C and Wilson, R.F (Eds). AOCS Press, New York. 1996.1:204-219.
[19] Q. Ma, X. Xu, Y. Gao, Q. Wang and J. Zhao. Optimisation of supercritical carbon dioxide extraction of lutein esters from marigold (Tagetes erecta L.) with soybean oil as a co-solvent. Int J Food Sci Technol. 2008. 43:pp.1763-1769.
[20] Mora-Pale, J.M. Perez-Munguia, S.Gonzalez-Mejia, J.C. Dordick and J.S. Barzana.. The lipase catalyzed hydrolysis of lutein diesters in nonaqueous media is favoured at extremely low water activities. Biotechnol Bioeng. 2007. 98(3).
[21] S. Naranjo-Modad, A. Lopez-Munguia, G. Vilarem, A. Gaset and E. Barzana.. Solubility of purified lutein diesters obtained from Tagetes erecta in supercritical CO2 and the effect of solvent modifiers. J. Agri. Food Chem. 2000.48: pp.5640- 5642.
[22] Rice-Evans, C.A., Sampson, J., Bramley, P.M and Holloway, D.E. 1997. Why do we expect carotenoids to be antioxidants in vivo? Free Radical Res. 26:381-398.
[23] A. J. Roodenburg, R. Leenen, K. H.Van het Hof, J. A. Weststrate and L. B. Tijburg. Amount of fat in the diet affects bioavailability of lutein esters but not of alpha-carotene, beta-carotene, and vitamin E in humans. Am J Clin Nutr. 2000.71:pp.1187-1193.
[24] G.J. Salter and D.B. Kell. Rapid determination using dielectric spectroscopy of the toxicity of organic solvents to intact cells. In: Biocatalysis in non-conventional media. Tramper, J., Vermue, M.H., Beeftinle, H.H and Von Stockar. U (Eds). Elsevier Science Publishers B.V. New York. 1992. 291-298.
[25] R.A. Sheldon. Chirotechnology-industrial synthesis of optically-active compounds. Marcel Dekker Ltd. New York. 1993. pp. 448.
[26] D.M. Small. Structure and Properties of Lipids. In: Biochemical and Physiological Aspects of Human Nutrition, Stipanuk, M.H (Ed). Philedelphia, PA: W.B. Saunders Company. 2000. pp. 43 - 62.
[27] J. Verghese.. Focus on xanthophylls from Tagetes erecta L the giant natural complex-I. Indian Spices. 1998. 33(4): pp.8-13.
[28] X. Xu, B. Shao, D. Zhou, S. Ye, Y. Wang and B. Chen. Process for the isolation and purification of xanthophylls crystals from plant oleoresin. 2007. US Patent 7,271,298, B2.
[29] H. Zorn, D.E. Breithaupt, M. Takenberg, W. Schwack and R.G. Berger. Enzymatic hydrolysis of carotenoid esters of marigold flowers (Tagetes erecta L.) and red paprika (Capsicum annuum L.) by commercial lipases and Pleurotus sapidus extracellular lipase. Enzyme Microb. Technol. 2003. 32. 623-628.