Preparation and application of epoxy–chitosan/alginate support in the immobilization of microbial lipases by covalent attachment

• Published in: Reactive & functional polymers Vol. 73; no. 1; pp. 160 - 167
• Main Authors: Mendes, Adriano A., de Castro, Heizir F., Andrade, Grazielle S.S., Tardioli, Paulo W., Giordano, Raquel de L.C.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01.01.2013; Elsevier
• Subjects: Applied sciences; Biological and medical sciences; Biotechnology; Catalytic properties; Covalent immobilization; Epoxy–chitosan/alginate; Ester synthesis; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Immobilization of enzymes and other molecules; Immobilization techniques; Methods. Procedures. Technologies; Microbial lipases; Natural polymers; Physicochemistry of polymers; Starch and polysaccharides; Catalytic properties; Ester synthesis; Covalent immobilization; Epoxy–chitosan/alginate; Microbial lipases; Support; Epichlorhydrin; Temperature effect; Immobilization; Triacylglycerol lipase; Esterases; Etherification; Butyric acid; Colloidal gel; Binding capacity; Enzymatic activity; Epoxy-chitosan/alginate; Enzyme; Alginates; Experimental study; Esterification; Carboxylic ester hydrolases; Polyelectrolyte; Chemical modification; Chitosan derivatives; Preparation; Covalent bond; Hydrolases; Oside polymer; Butanol; Chitosan; Immobilized enzyme
• ISSN: 1381-5148
• DOI: 10.1016/j.reactfunctpolym.2012.08.023

The aim of this work was the preparation and application of highly hydrophobic epoxy–chitosan/alginate as a support to immobilize microbial lipases from Thermomyces lanuginosus commercially available as Lipolase® (TLL1) and Lipex® 100L (TLL2) and Pseudomonas fluorescens (PFL). The catalytic properties of the biocatalysts were assayed in olive oil hydrolysis and butyl butyrate synthesis. The results indicated that 12h was enough for TLL1 to be immobilized on the support. Covalent attachment of TLL1 turned biocatalysts highly active and around 6-fold more stable than free lipase. Based on the results, a time of incubation of 24h was selected for further studies about the maximum immobilized protein amount and butyl butyrate synthesis. Maximum protein loading immobilized was found to be 25.4mgg−1support for TLL1, followed by TLL2 (20.5mgg−1) and PFL (15.5mgg−1) offering 80mgproteing−1support. The immobilization of TLL1 and TLL2 resulted in highly active biocatalysts (around 1300IUg−1gel), almost fivefold higher than PFL (272.4IUg−1gel). In butyl butyrate synthesis, PFL showed similar activity to TLL1 and TLL2 derivatives, up to 60mmolL−1. The biocatalysts displayed high activity after five successive cycles, retaining around 95% of the initial activity.