title: Conversion of quinate to 3-dehydroshikimate by Ca-alginate-immobilized membrane of Gluconobacter oxydans IFO 3244 and subsequent asymmetric reduction of 3-dehydroshikimate to shikimate by immobilized cytoplasmic NADP-shikimate dehydrogenase.
abstract: The membrane fraction of Gluconobacter oxydans IFO 3244, involving membrane-bound quinoprotein quinate dehydrogenase and 3-dehydroquinate dehydratase, was immobilized into Ca-alginate beads. The Ca-alginate-immobilized bacterial membrane catalyzed a sequential reaction of quinate oxidation to 3-dehydroquinate and its spontaneous conversion to 3-dehydroshikimate under neutral pH. An almost 100% conversion rate from quinate to 3-dehydroshikimate was observed. NADP-Dependent cytoplasmic enzymes from the same organism, shikimate dehydrogenase and D-glucose dehydrogenase, were immobilized together with different carriers as an asymmetric reduction system forming shikimate from 3-dehydroshikimate. Blue Dextran 2000, Blue Dextran-Sepharose-4B, DEAE-Sephadex A-50, DEAE-cellulose, and hydroxyapatite were effective carriers of the two cytoplasmic enzymes, and the 3-dehydroshikimate initially added was converted to shikimate at 100% yield. The two cytoplasmic enzymes showed strong affinity to Blue Dextran 2000 and formed a soluble form of immobilized catalyst having the same catalytic efficiency as that of the free enzymes. This paper may be the first one on successful immobilization of NAD(P)-dependent dehydrogenases.
authors.affiliation: Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan. osao@yamaguchi-u.ac.jp
authors.forename: Osao
authors.initials: O
authors.lastname: Adachi
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Ano
authors.forename: Emiko
authors.initials: E
authors.lastname: Shinagawa
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Yakushi
authors.forename: Kazunobu
authors.initials: K
authors.lastname: Matsushita
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D000464:Alginates / Q000737:chemistry*; D055162:Biocatalysis; D002462:Cell Membrane / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology*; D003911:Dextrans / Q000378:metabolism; D017886:Durapatite / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005285:Fermentation; D020600:Gluconobacter oxydans / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D007474:Ion Exchange; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D011801:Quinic Acid / Q000378:metabolism*; D012765:Shikimic Acid / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D000464:Alginates; D003911:Dextrans; D004800:Enzymes, Immobilized; D006603:Hexuronic Acids; D011801:Quinic Acid; C096387:3-dehydroshikimate; D012765:Shikimic Acid; D009249:NADP; D020723:Glucuronic Acid; C017080:blue dextran; D017886:Durapatite; D000429:Alcohol Oxidoreductases; C022641:Shikimate dehydrogenase