ENZYMATIC TRANSESTERIFICATION OF FLAVONOIDS Jazmín Pineda1, Arturo Navarro1, Ernesto Sanchez 2, Liliana Hernández2 1 Faculty of Chemistry, Department of Food and Biotechnology, National Autonomous University of Mexico (UNAM), [email protected] 2 Department of Biological System, UAM-Xochimilco, [email protected] Keywords: Flavonoids; Lipase; Esterification Figure1. TLC of transesterification of naringin with methyl ester of fatty acids INTRODUCTION Has been evidence that flavonoids have beneficial health properties, however, the use of flavonoids in several domains is limited by their low stability and solubility in the fatty phase. One solution to improve their hydrophobic nature is the esterification; this chemical reaction leads to a mixture of products (1). To overcome this problem, enzymatic catalysis is an option for its regioselectivity. The lipases are capable of promoting the synthesis of the ester, when the reaction is carried out in a low water activity. The aim of this work is to transform flavonoids (naringin and rutin) by way transesterification reaction with fatty acids from vegetable oils and their methyl esters, activated with lipase from C. antarctica RESULTS AND DISCUSSION Naringin and rutin were enzymatically acylated with different fatty acids as acyl donors (in this case they were added as oil) Table 1.Transesterification of flavonoids with different acylating agents Acyl donor Coconut oil Nutmeg Oil Olive oil Almond oil Grapeseed oil Pumpkin oil Linseed oil Chia oil Castor oil Fatty acid in the oil C12:0 C14:0 C18:1 C18:1 C18:2 Acyl acceptor Naringin Rutin + + + + + + + + + + 1 .Methyl ester of Coconut oil 2. Methyl ester of Nutmeg Oil 3. Methyl ester of Olive oil 4. Methyl ester of Almond oil 5. Methyl ester of Grapeseed oil 6. Methyl ester of Pumpkin oil 7. Methyl ester of Linseed oil 8. Methyl ester of Chia oil 9. Methyl ester of Castor oil 10. Naringin The reaction was carried out with all the methyl esters (Figure 1) and no significant difference was observed in the rate of conversion reactions with respect to oils. With C18 fatty acids, yields are in the same range of magnitude, between 50 and 60% with naringin, and between 40 and 50% with rutin. For fatty acids C12C14 yields are between 20-30% for both flavonoids. Higher conversion yields of naringin and rutin were obtained with aliphatic acids having high carbon chain length (C18) as reported by other authors (2). The fatty acid esters of rutin and naringin were 13 1 purified and characterized by C y H NMR. The results suggest the presence of an ester bond on the C-6 of the glucose moiety of naringin molecule. While the acylation of rutin took place precisely on the secondary OH in the C-4 of the rhamnose moiety in concordance with other studies (3). CONCLUSION Lipase of C. antarctica is capable of carry out the esterification reaction of flavonoids naringin and rutin with fatty acids used. With the purification of the products and their identification is observed regioselectivity of the enzyme reaction. ACKNOWLEDGEMENTS C18:2 C18:3 C18:3 C18:1 + + + + + + + + + Reaction – No reaction For naringin and rutin was observed that the transesterification reaction was carried out with all the acylating agents that were tested (Table 1) to give a single reaction product in all cases. The methyl esters were obtained by enzymatic reaction and then used for the esterification of the flavonoid. The authors are grateful to the CONACYT-2012CB180128. REFERENCES [1]. Chebil, L.; Humeau, C.; Falcimaigne, a.; Engasser, J.; Ghoul, M. Review Enzymatic acylation of flavonoids. Process Biochemistry, 2006, 2237-2251 [2]. Ardhaoui, M.; Falcimaigne, A.; Ognier, S.; Engasser,J.; Moussoub, P.; Pauly , G.; Ghoul, M. Effect of acyl donor chain length and substitutions pattern on the enzymatic acylation of flavonoids. Journal of Biotechnology, 2004, 110, 265-271 [3]. Kontogianni, A.; Skouridou, V.; Sereti, V.; Stamatis, H.; Kolisis, F.; Lipase-catalyzed esterification of rutin and naringin with fatty acids of medium carbon chain. Journal of Molecular Catalysis, 2003, 23, 59-62. VII Workshop on Biocatalysis and Biotransformations and 1o Simposio Latinoamericano de Biocatalisis y Biotransformaciones