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PublicationBiotechnology Progress

Optimized butyl butyrate synthesis catalyzed by Thermomyces lanuginosus lipase

Authors:Roberto Fernandez-Lafuente, Cristina Garcia-Galan
Groups of research:Optimización de biocatalizadores y bioprocesos enzimáticos
Andréa B. Martins1a, John L. R Friedrich1a, Cristina Garcia-Galan2, Roberto Fernandez-Lafuente2, Marco A. Z. Ayub1b, Rafael C. Rodrigues1a,*
1aBiocatalysis and Enzyme Technology Lab and 1bBiochemical Engineering Lab (BiotecLab), Institute of Food Science and Technology, Federal University of Rio Grande do Sul State, Av. Bento Gonçalves, 9500, P.O. Box 15090, ZC 91501-970, Porto Alegre, RS, Brazil.
2Department of Biocatalysis, ICP - CSIC. Campus UAM-CSIC. Cantoblanco, ZC 28049, Madrid, Spain.

Butyl butyrate is an ester present in pineapple flavor, which is very important for the food and beverages industries. In this work, it was performed the optimization of the reaction of butyl butyrate synthesis catalyzed by the immobilized lipase Lipozyme TL-IM. n-Hexane was selected as the most appropriate solvent. Other reaction parameters such as temperature, substrate molar ratio, biocatalyst content and added water, and their responses measured as conversion yields, were evaluated using a fractional factorial design (FFD), followed by a central composite design (CCD) and response surface methodology. In the fractional design 24-1, the four variables were tested and temperature and biocatalyst content were statistically significant and then used for optimization on CCD. The optimal conditions for butyl butyrate synthesis were found to be 48 ºC; substrate molar ratio 3:1 (butanol:butyric acid); biocatalyst content of 40 % of acid mass. Under these conditions, over 90 % of conversion was obtained in 2 h. Enzyme reuse was tested by washing the biocatalyst with n-hexane or by direct reuse. The direct reuse produced a rapid decrease on enzyme activity, while washing with n-hexane allowed reusing the enzyme for 5 reactions cycles keeping approximately 85 % of its activity.
Keywords:Butyl butyrate; lipase; esterification; experimental design; enzyme reuse
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