title: Design of biocompatible immobilized Candida rugosa lipase with potential application in food industry.
abstract: BACKGROUND
  Biocatalysts are a promising alternative for the production of natural flavor compounds. Candida rugosa lipase (CRL) is a particularly important biocatalyst owing to its remarkable efficiency in both hydrolysis and synthesis. However, additional stabilization is necessary for successful industrial implementation. This study presents an easy and time-saving method for immobilizing this valuable enzyme on hydroxyapatite (HAP), a biomaterial with high protein-binding capacity.
  
  
  RESULTS
  Targeted immobilized CRL was obtained in high yield of ≥98%. Significant lipase stabilization was observed upon immobilization: at 60 °C, immobilized lipase (HAP-CRL) retained almost unchanged activity after 3 h, while free CRL lost 50% of its initial activity after only 30 min. The same trend was observed with tested organic solvents. Methanol and hexane had the most pronounced effect: after 3 h, only HAP-CRL was stable and active, while CRL was completely inactivated. The practical value of the prepared catalyst was tested in the synthesis of the aroma ester methyl acetate in hexane. Reaction yields were 2.6 and 52.5% for CRL and HAP-CRL respectively.
  
  
  CONCLUSIONS
  This research has successfully combined an industrially prominent biocatalyst, CRL, and a biocompatible, environmentally suitable carrier, HAP, into an immobilized preparation with improved catalytic properties. The obtained CRL preparation has excellent potential for the food and flavor industries, major consumers in the global enzyme market. © 2016 Society of Chemical Industry.
authors.affiliation: Innovation Center, Faculty of Chemistry, University of Belgrade, 11000, Belgrade, Serbia.
authors.forename: Jovana
authors.initials: J
authors.lastname: Trbojević Ivić
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Belgrade, 11000, Belgrade, Serbia.
authors.forename: Dušan
authors.initials: D
authors.lastname: Veličković
authors.affiliation: Department of Molecular Biology and Biochemistry, University of California Irvine, Irvine, CA, 92697, USA.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Dimitrijević
authors.affiliation: Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, 11000, Belgrade, Serbia.
authors.forename: Dejan
authors.initials: D
authors.lastname: Bezbradica
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Belgrade, 11000, Belgrade, Serbia.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Dragačević
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Belgrade, 11000, Belgrade, Serbia.
authors.forename: Marija
authors.initials: M
authors.lastname: Gavrović Jankulović
authors.affiliation: Division of Experimental Therapeutics, Department of Medicine, Columbia University, New York, NY, 10032, USA.
authors.forename: Nenad
authors.initials: N
authors.lastname: Milosavić
mesh_terms: D000085:Acetates / Q000378:metabolism; D055162:Biocatalysis; D001709:Biotechnology; D002175:Candida / Q000201:enzymology*; D017886:Durapatite / Q000737:chemistry; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D019649:Food Industry; D006586:Hexanes / Q000737:chemistry; D006868:Hydrolysis; D008049:Lipase / Q000737:chemistry* / Q000378:metabolism; D008422:Materials Testing; D000432:Methanol / Q000737:chemistry
chemical_list: D000085:Acetates; D004800:Enzymes, Immobilized; D006586:Hexanes; D017886:Durapatite; D008049:Lipase; C046923:methyl acetate; D000432:Methanol
keywords: Candida rugosa lipase; hydroxyapatite; immobilization; methyl acetate; organic solvents