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An efficient method for enzymatic purification of cis-9,trans-11 isomer of conjugated linoleic acid
- Niezgoda, Natalia, Wawrzeńczyk, Czesław
- Journal of Molecular Catalysis. B, Enzymatic 2014 v.100 pp. 40-48
- Candida cylindracea, alcohols, carboxylic ester hydrolases, catalytic activity, conjugated linoleic acid, crystallization, enzymatic treatment, esterification, isomers, pH, purification methods, temperature
- A method for purification of cis-9,trans-11 isomer of conjugated linoleic acid (CLA) is described. Lipase from Candida cylindracea (CCL) was highly selective towards cis-9,trans-11 isomer and was chosen to catalyze esterification of a mixture of two CLA isomers. The effect of various parameters on the esterification process of an almost equimolar mixture of cis-9,trans-11 (44.0%), trans-10,cis-12 (42.5%) and other CLA isomers (13.5%) was studied. The influence of alcohol substrate, CLA/alcohol molar ratio, lipase amount, reaction temperature, pH and time of the reaction on the total esterification and cis-9,trans-11 isomer purity in ester fraction were examined. The highest isomeric excess of cis-9,trans-11 isomer on trans-10,cis-12 CLA and good esterification degree was observed when the esterification condition were as follows: CLA/2-propanol molar ratio, 1:5; water, 200% wt of CLA; CCL, 24μg of CCL/mg CLA; temperature 30̊C; time of reaction 6h and when mixture enriched in cis-9,trans-11 CLA was used as a substrate for esterification. A preparative scale purification combining crystallization and one step enzymatic process were made to afford purified cis-9,trans-11 (94.2%) with total recovery of 31%.