title: Biotechnological production of vanillin using immobilized enzymes.
abstract: Vanillin is an important and popular plant flavor, but the amount of this compound available from plant sources is very limited. Biotechnological methods have high potential for vanillin production as an alternative to extraction from plant sources. Here, we report a new approach using immobilized enzymes for the production of vanillin. The recently discovered oxygenase Cso2 has coenzyme-independent catalytic activity for the conversion of isoeugenol and 4-vinylguaiacol to vanillin. Immobilization of Cso2 on Sepabeads EC-EA anion-exchange carrier conferred enhanced operational stability enabling repetitive use. This immobilized Cso2 catalyst allowed 6.8mg yield of vanillin from isoeugenol through ten reaction cycles at a 1mL scale. The coenzyme-independent decarboxylase Fdc, which has catalytic activity for the conversion of ferulic acid to 4-vinylguaiacol, was also immobilized on Sepabeads EC-EA. We demonstrated that the immobilized Fdc and Cso2 enabled the cascade synthesis of vanillin from ferulic acid via 4-vinylguaiacol with repetitive use of the catalysts. This study is the first example of biotechnological production of vanillin using immobilized enzymes, a process that provides new possibilities for vanillin production.
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Engineering, Waseda University, 3-4-1 Ohkubo, Shinjuku-ku, Tokyo 169-8555, Japan; Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan. Electronic address: tfuruya@rs.tus.ac.jp.
authors.forename: Toshiki
authors.initials: T
authors.lastname: Furuya
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Engineering, Waseda University, 3-4-1 Ohkubo, Shinjuku-ku, Tokyo 169-8555, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Kuroiwa
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Engineering, Waseda University, 3-4-1 Ohkubo, Shinjuku-ku, Tokyo 169-8555, Japan. Electronic address: kkino@waseda.jp.
authors.forename: Kuniki
authors.initials: K
authors.lastname: Kino
mesh_terms: D001547:Benzaldehydes / Q000737:chemistry / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D002262:Carboxy-Lyases / Q000737:chemistry / Q000378:metabolism*; D002384:Catalysis; D003067:Coenzymes; D003373:Coumaric Acids / Q000737:chemistry / Q000378:metabolism; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D005054:Eugenol / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D006139:Guaiacol / Q000031:analogs & derivatives / Q000378:metabolism; D010105:Oxygenases / Q000737:chemistry / Q000378:metabolism
chemical_list: D001547:Benzaldehydes; D003067:Coenzymes; D003373:Coumaric Acids; D004800:Enzymes, Immobilized; D005054:Eugenol; C036643:isoeugenol; D006139:Guaiacol; C014245:4-vinylguaiacol; C004999:ferulic acid; C100058:vanillin; D010105:Oxygenases; D002262:Carboxy-Lyases
keywords: Biocatalysis; Cascade reaction; Ferulic acid; Immobilized enzyme; Vanillin