Improved production of butyl butyrate with lipase from Thermomyces lanuginosus immobilized on styrene–divinylbenzene beads

• Published in: Bioresource technology Vol. 134; pp. 417 - 422
• Main Authors: Martins, Andréa B., Friedrich, John L.R., Cavalheiro, Jhonnattas C., Garcia-Galan, Cristina, Barbosa, Oveimar, Ayub, Marco A.Z., Fernandez-Lafuente, Roberto, Rodrigues, Rafael C.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Ltd 01-04-2013; Elsevier
• Subjects: Ascomycota - cytology; Ascomycota - drug effects; Ascomycota - enzymology; Biocatalysis - drug effects; Biological and medical sciences; Biotechnology - methods; Butyl butyrate; Butyrates - metabolism; Butyrates - pharmacology; Cells, Immobilized - cytology; Cells, Immobilized - drug effects; Cells, Immobilized - enzymology; Electrophoresis, Polyacrylamide Gel; Esterification; Esterification - drug effects; Fundamental and applied biological sciences. Psychology; Interfacial activated lipases; Kinetics; Lipase - metabolism; Microspheres; Styrene - pharmacology; Styrene–divinylbenzene beads; Substrate Specificity - drug effects; Temperature; TLL; Vinyl Compounds - pharmacology; Water - pharmacology; Butyl butyrate; Styrene–divinylbenzene beads; TLL; Interfacial activated lipases; Esterification; Fungi; Styrene-divinylbenzene beads; Enzyme; Styrene; Thermomyces lanuginosus; Fungi Imperfecti; Butyrate; Lipolytic enzyme
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2013.02.052

► TLL was immobilized on styrene-divinylbenzene beads (MCI-TLL). ► MCI-TLL was compared to Lipozyme TL-IM in butyl butyrate synthesis. ► MCI-TLL presented two-times more protein than Lipozyme TL-IM. ► The prepared biocatalyst was stable to higher concentration of butyric acid. ► MCI-TLL showed productivity 4-times higher than Lipozyme TL-IM.
Two immobilized preparations from Thermomyces lanuginosus lipase (TLL) were compared in the synthesis of butyl butyrate. The commercial Lipozyme TL-IM, and TLL immobilized on styrene–divinylbenzene beads (MCI-TLL) were tested in the esterification reaction using n-hexane as solvent. The variables temperature (30–60°C), substrate molar ratio (1:1 to 5:1), added water (0–1%), and biocatalyst content (3–40%) were evaluated in terms of initial reaction rate for each biocatalyst. SDS–PAGE analysis revealed that MCI-TLL had an immobilized enzymatic load twice as high as Lipozyme TL-IM, but with an activity 3-fold higher. MCI-TLL presented high initial reaction rates up to 1.0M butyric acid, while Lipozyme TL-IM showed a decrease in its activity above 0.5M. Moreover, MCI-TLL allowed a productivity of 14.5mmolg−1h−1, while Lipozyme TL-IM 3.2mmolg−1h−1, both by mass of biocatalyst.