title: Purification and characterization of d-allulose 3-epimerase derived from Arthrobacter globiformis M30, a GRAS microorganism.
abstract: An enzyme that catalyzes C-3 epimerization between d-fructose and d-allulose was found in Arthrobacter globiformis strain M30. Arthrobacter species have long been used in the food industry and are well-known for their high degree of safety. The enzyme was purified by ion exchange and hydrophobic interaction chromatographies and characterized as a d-allulose 3-epimerase (d-AE). The molecular weight of the purified enzyme was estimated to be 128 kDa with four identical subunits. The enzyme showed maximal activity and thermostability in the presence of Mg2+. The optimal pH and temperature for enzymatic activity were 7.0-8.0 and 70°C, respectively. The enzyme was immobilized to ion exchange resin whereupon it was stable for longer periods than the free enzyme when stored at below 10°C. In the column reaction, the enzyme activity also maintained stability for more than 4 months. Under these conditions, 215 kg of d-allulose produced per liter immobilized enzyme, and this was the highest production yield of d-allulose reported so far. These highly stable properties suggest that this enzyme represents an ideal candidate for the industrial production of d-allulose.
authors.affiliation: Rare Sugar Research Center, Kagawa University, 2393 Ikenobe, Miki 761-0795, Kagawa, Japan.
authors.forename: Akihide
authors.initials: A
authors.lastname: Yoshihara
authors.affiliation: Faculty of Agriculture, Kagawa University, 2393 Ikenobe, Miki 761-0795, Kagawa, Japan.
authors.forename: Taro
authors.initials: T
authors.lastname: Kozakai
authors.affiliation: Matsutani Chemical Industry Co., Ltd, 5-3 Kita-Itami, Itami 664-8508, Hyogo, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Shintani
authors.affiliation: Matsutani Chemical Industry Co., Ltd, 5-3 Kita-Itami, Itami 664-8508, Hyogo, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Matsutani
authors.affiliation: Matsutani Chemical Industry Co., Ltd, 5-3 Kita-Itami, Itami 664-8508, Hyogo, Japan.
authors.forename: Kouhei
authors.initials: K
authors.lastname: Ohtani
authors.affiliation: Matsutani Chemical Industry Co., Ltd, 5-3 Kita-Itami, Itami 664-8508, Hyogo, Japan.
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Iida
authors.affiliation: Faculty of Agriculture, Kagawa University, 2393 Ikenobe, Miki 761-0795, Kagawa, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Akimitsu
authors.affiliation: Faculty of Agriculture, Kagawa University, 2393 Ikenobe, Miki 761-0795, Kagawa, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Izumori
authors.affiliation: Matsutani Chemical Industry Co., Ltd, 5-3 Kita-Itami, Itami 664-8508, Hyogo, Japan. Electronic address: pushpa-kiran-gullapalli@matsutani.co.jp.
authors.forename: Pushpa Kiran
authors.initials: PK
authors.lastname: Gullapalli
mesh_terms: D001173:Arthrobacter / Q000737:chemistry / Q000201:enzymology*; D004795:Enzyme Stability; D005632:Fructose / Q000096:biosynthesis / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D060847:Metabolic Engineering; D008970:Molecular Weight; D019998:Racemases and Epimerases / Q000032:analysis* / Q000302:isolation & purification* / Q000378:metabolism*; D013696:Temperature
chemical_list: C003243:psicose; D005632:Fructose; D019998:Racemases and Epimerases
keywords: Arthrobacter globiformis M30; Immobilized d-AE; Magnesium; d-Allulose; d-Allulose 3-epimerase