title: Immobilization of a Cutinase from Fusarium oxysporum and Application in Pineapple Flavor Synthesis.
abstract: In the present study, the immobilization of a cutinase from Fusarium oxysporum was carried out as cross-linked enzyme aggregates. Under optimal immobilization conditions, acetonitrile was selected as precipitant, utilizing 9.4 mg protein/mL and 10 mM glutaraldehyde as cross-linker. The immobilized cutinase (imFocut5a) was tested in isooctane for the synthesis of short-chain butyrate esters, displaying enhanced thermostability compared to the free enzyme. Pineapple flavor (butyl butyrate) synthesis was optimized, leading to a conversion yield of >99% after 6 h, with an initial reaction rate of 18.2 mmol/L/h. Optimal reaction conditions were found to be 50 °C, a vinyl butyrate/butanol molar ratio of 3:1, vinyl butyrate concentration of 100 mM, and enzyme loading of 11 U. Reusability studies of imFocut5a showed that after four consecutive runs, the reaction yield reaches 54% of the maximum. The efficient bioconversion offers a sustainable and environmentally friendly process for the production of "natural" aroma compounds essential for the food industry.
authors.affiliation: Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens , 9 Heroon Polytechniou Street, Zographou Campus, 15780 Athens, Greece.
authors.forename: Efstratios
authors.initials: E
authors.lastname: Nikolaivits
authors.affiliation: Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens , 9 Heroon Polytechniou Street, Zographou Campus, 15780 Athens, Greece.
authors.forename: Georgios
authors.initials: G
authors.lastname: Makris
authors.affiliation: Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens , 9 Heroon Polytechniou Street, Zographou Campus, 15780 Athens, Greece.
authors.forename: Evangelos
authors.identifier: http://orcid.org/0000-0003-0078-5904
authors.initials: E
authors.lastname: Topakas
mesh_terms: D031230:Ananas / Q000737:chemistry; D055162:Biocatalysis; D002087:Butyrates / Q000138:chemical synthesis* / Q000737:chemistry; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005421:Flavoring Agents / Q000737:chemistry*; D005656:Fungal Proteins / Q000737:chemistry*; D005670:Fusarium / Q000201:enzymology*
chemical_list: D002087:Butyrates; D004800:Enzymes, Immobilized; D005421:Flavoring Agents; D005656:Fungal Proteins; C022793:n-butyl n-butyrate; D002265:Carboxylic Ester Hydrolases; C019952:cutinase
keywords: biocatalysis; cross-linked enzyme aggregates; cutinase; flavor esters; immobilization; transesterification