title: Immobilization of Candida antarctica lipase B onto Purolite®MN102 and its application in solvent-free and organic media esterification.
abstract: The aim of this study was to develop simple and efficient method for immobilization of Candida antarctica lipase B onto hydrophobic anion exchange resin Purolite®MN102 and to apply immobilized catalyst for the enzymatic synthesis of two valuable esters-isoamyl acetate and L-ascorbyl oleate. At optimized conditions (1 M phosphate buffer pH = 7, 7 h at 25 °C, and 18.75 mg of offered proteins g-1of support), immobilized lipase with hydrolytic activity of 888.4 p-nitrophenyl butyrate units g-1was obtained. Afterwards, preparation was applied for the solvent-free synthesis of isoamyl acetate from triacetin and isoamyl alcohol. At 75 °C, 1 M of isoamyl alcohol, and 6 mg ml-1of enzyme 100 % yield was achieved in 6 h, while at prolonged reaction times, complete conversion was enabled even at lower temperatures, lower lipase loadings, and higher substrate concentrations. After 15 consecutive reuses (60 h), activity of catalyst dropped to 50 % of its initial value and total amount of 1.31 mol (170.55 g) of ester with 1 g of biocatalyst was produced. Even higher operational stability of lipase (25 % loss of activity in 200 h) was observed in the synthesis of L-ascorbyl oleate performed in organic solvent (t-butanol). Multiple use of one batch of immobilized biocatalyst in both cases led to a significant process cost reduction and substantial increment of corresponding productivities.
authors.affiliation: Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000, Belgrade, Serbia.
authors.forename: Marija
authors.initials: M
authors.lastname: Ćorović
authors.affiliation: Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000, Belgrade, Serbia.
authors.forename: Mladen
authors.initials: M
authors.lastname: Mihailović
authors.affiliation: Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000, Belgrade, Serbia.
authors.forename: Katarina
authors.initials: K
authors.lastname: Banjanac
authors.affiliation: Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000, Belgrade, Serbia.
authors.forename: Milica
authors.initials: M
authors.lastname: Carević
authors.affiliation: Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000, Belgrade, Serbia.
authors.forename: Ana
authors.initials: A
authors.lastname: Milivojević
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12, 11000, Belgrade, Serbia.
authors.forename: Nenad
authors.initials: N
authors.lastname: Milosavić
authors.affiliation: Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000, Belgrade, Serbia. dbez@tmf.bg.ac.rs.
authors.forename: Dejan
authors.initials: D
authors.lastname: Bezbradica
mesh_terms: D000837:Anion Exchange Resins / Q000737:chemistry*; D002175:Candida / Q000201:enzymology; D002384:Catalysis; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005656:Fungal Proteins / Q000737:chemistry*; D008049:Lipase / Q000737:chemistry*
chemical_list: D000837:Anion Exchange Resins; D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D008049:Lipase; C431349:lipase B, Candida antarctica
keywords: Candida antarctica lipase B; Esterification; Immobilization; Isoamyl acetate; L-Ascorbyl oleate