TY - JOUR
T1 - Characterization of a bacterial pyranose 2-oxidase from Arthrobacter siccitolerans
AU - Mendes, Sónia
AU - Banha, Catarina
AU - Madeira, Joaquim
AU - Santos, Diana
AU - Miranda, Vanessa
AU - Manzanera, Maximino
AU - Ventura, M.R.
AU - van Berkel, Willem J.H.
AU - Martins, Lígia O.
PY - 2016/11
Y1 - 2016/11
N2 - In this study we provide the first biochemical characterization of a bacterial pyranose 2-oxidase (AsP2Ox) from Arthrobacter siccitolerans. The enzyme catalyzes the oxidation of several aldopyranoses at the C-2 position, coupling it to the reduction of dioxygen to hydrogen peroxide. Pyranose 2-oxidases belong to the glucose-methanol-choline oxidoreductase family. A structural model based on the known X-ray structure of P2Ox from Phanerochaete chrysosporium supports that AsP2Ox shares structural features with well-characterized fungal P2Oxs. The gene coding for AsP2Ox was cloned and heterologously expressed in Escherichia coli. The purified recombinant enzyme is a 64-kDa monomer containing a non-covalently bound flavin adenine dinucleotide (FAD) cofactor, distinct features as compared with fungal counterparts that are ∼ 270kDa homotetramers with covalent-linked FAD. AsP2Ox exhibits a redox potential of -50mV, an optimum temperature of 37C and an optimum pH at 6.5. AsP2Ox oxidizes d-glucose at the highest efficiency, using additionally d-galactose, d-xylose, l-arabinose and d-ribose as electron donors, coupling their oxidation to the reduction of both dioxygen and 1,4-benzoquinone. AsP2Ox shows a relatively low thermal stability with a melting temperature (T m) of 43C and a half-life (t1/2) at 40C of 25min. This work expands the repertoire of bacterial oxidoreductases with importance in biotechnological and diagnostic applications.
AB - In this study we provide the first biochemical characterization of a bacterial pyranose 2-oxidase (AsP2Ox) from Arthrobacter siccitolerans. The enzyme catalyzes the oxidation of several aldopyranoses at the C-2 position, coupling it to the reduction of dioxygen to hydrogen peroxide. Pyranose 2-oxidases belong to the glucose-methanol-choline oxidoreductase family. A structural model based on the known X-ray structure of P2Ox from Phanerochaete chrysosporium supports that AsP2Ox shares structural features with well-characterized fungal P2Oxs. The gene coding for AsP2Ox was cloned and heterologously expressed in Escherichia coli. The purified recombinant enzyme is a 64-kDa monomer containing a non-covalently bound flavin adenine dinucleotide (FAD) cofactor, distinct features as compared with fungal counterparts that are ∼ 270kDa homotetramers with covalent-linked FAD. AsP2Ox exhibits a redox potential of -50mV, an optimum temperature of 37C and an optimum pH at 6.5. AsP2Ox oxidizes d-glucose at the highest efficiency, using additionally d-galactose, d-xylose, l-arabinose and d-ribose as electron donors, coupling their oxidation to the reduction of both dioxygen and 1,4-benzoquinone. AsP2Ox shows a relatively low thermal stability with a melting temperature (T m) of 43C and a half-life (t1/2) at 40C of 25min. This work expands the repertoire of bacterial oxidoreductases with importance in biotechnological and diagnostic applications.
KW - Carbohydrate chemistry
KW - Flavoprotein
KW - Glucose-methanol-choline family of oxidoreductases
KW - Hydrogen peroxide forming enzymes
U2 - 10.1016/j.molcatb.2016.11.005
DO - 10.1016/j.molcatb.2016.11.005
M3 - Article
AN - SCOPUS:85007560471
SN - 1381-1177
VL - 133
SP - S34-S43
JO - Journal of Molecular Catalysis. B, Enzymatic
JF - Journal of Molecular Catalysis. B, Enzymatic
IS - suppl. 1
ER -