Goal-oriented screening method for carbohydrate oxidases produced by filamentous fungi

Danneel, Hans-Jürgen; Ullrich, Michael; Giffhorn, Friedrich

doi:10.1016/0141-0229(92)90053-q

Cited by 26 publications

(22 citation statements)

References 27 publications

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“…The organism used in this study was a filamentous fungus isolated from forest soil near Gottingen (Germany, Danneel et al, 1992). The fungus was classified as the basidiomycete Peniophora gigantea by the German Culture Collection (DSM), Braunschweig.…”

Section: Materials and Methods Organism And Growthmentioning

confidence: 99%

“…The most prominent process is the chemoenzymic transformation of D-glucose via D-glucosone (~-arabino-hexos-2-ulose) as the key intermediate of the industrial production of D-fructose (Liu et al, 1983). Since relatively little information exists on the biochemical and biophysical properties of pyranose oxidases, we have isolated and characterized a corresponding enzyme from a basidiomycete obtained from a goal-orientated screening device for carbohydrate oxidases produced by microorganisms from soil (Danneel et al, 1992).…”

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confidence: 99%

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Purification and characterization of a pyranose oxidase from the basidiomycete Peniophora gigantea and chemical analyses of its reaction products

Danneel

Rössner

Zeeck

et al. 1993

European Journal of Biochemistry

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A pyranose oxidase was isolated from mycelium extracts of the basidiomycete Peniophora gigantea. This enzyme was purified 104-fold to apparent homogeneity with a yield of about 75% by steps involving fractionated ammonium sulphate precipitation, chromatography on DEAE-Sephacel, Sephacryl S 300, S Sepharose and Q Sepharose. The native pyranose oxidase has a relative molecular mass (MJ of 322800 It 18300 as determined on the basis of its Stokes' radius (rs = 6.2 nm) and sedimentation coefficient = 10.6), dynamic light-scattering experiments, gradient-gel electrophoresis and cross-linking studies. SDSPAGE resulted in one single polypeptide band of Mr76000 indicating that the enzyme consists of four subunits of identical size. The pyranose oxidase was shown to be an extremely stable glycoprotein with an isoelectric point of pH 5.3. It contains covalently bound FAD with an estimated stoichiometry of 3.6 molecules FAD/molecule enzyme. Pyranose oxidase was active with the substrates D-glucose, D-xylose, L-sorbose, D-galactose, methyl P-D-glucoside, maltose and D-fucose. Regioselective oxidation of D-glucose, L-sorbose and D-xylose to 2-keto-~-glucose, 5-keto-~-fructose and 2-keto-D-xylose, was demonstrated by identifying the reaction products by mass spectroscopy I3C-NMR spectroscopy and 'H-NMR spectroscopy after purification and derivatization. The pH optimum of the pyranose oxidase was in the range pH 6.0-6.5 in 0.1 M potassium phosphate, and its activation energy ( A H") for the conversion of D-glucose was 34.6 kJ/mol. The reactions with the sugars exhibited Michaelis-Menten kinetics, and the K, values determined for D-glucose, L-sorbose, D-xylose and oxygen were 1.1 mM, 50.0 mM, 29.4 mM and 0.65 mM, respectively. The activity of pyranose oxidase was only slightly affected by chelating reagents, thiol reagents, reducing reagents and bivalent cations each at 1 mM.

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Section: Materials and Methods Organism And Growthmentioning

confidence: 99%

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confidence: 99%

Purification and characterization of a pyranose oxidase from the basidiomycete Peniophora gigantea and chemical analyses of its reaction products

Danneel

Rössner

Zeeck

et al. 1993

European Journal of Biochemistry

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“…It has been proposed that the main metabolic function of pyranose oxidase is as a constituent of the ligninolytic system of white rot fungi that provides peroxidases with H 2 O 2 . An additional function could be reduction of quinones, key intermediates that are formed during mineralization of lignin.The enzyme pyranose oxidase (P2O) (pyranose:oxygen 2-oxidoreductase; EC 1.1.3.10), which catalyzes the oxidation of several aldopyranoses at position C-2 to yield the corresponding 2-ketoaldoses (aldos-2-uloses, osones), is widely distributed among wood-degrading basidiomycetes (14,32,44). It has been purified and characterized from several microorganisms, including Phanerochaete chrysosporium (2, 45), Phlebiopsis gigantea (39), Pleurotus ostreatus (40), Polyporus obtusus (28), Trametes (Coriolus) versicolor (35), and unidentified basidiomycete no.…”

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“…The enzyme pyranose oxidase (P2O) (pyranose:oxygen 2-oxidoreductase; EC 1.1.3.10), which catalyzes the oxidation of several aldopyranoses at position C-2 to yield the corresponding 2-ketoaldoses (aldos-2-uloses, osones), is widely distributed among wood-degrading basidiomycetes (14,32,44). It has been purified and characterized from several microorganisms, including Phanerochaete chrysosporium (2,45), Phlebiopsis gigantea (39), Pleurotus ostreatus (40), Polyporus obtusus (28), Trametes (Coriolus) versicolor (35), and unidentified basidiomycete no.…”

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Purification and Characterization of Pyranose Oxidase from the White Rot Fungus Trametes multicolor

Leitner¹,

Volc

Haltrich³

2001

Appl Environ Microbiol

146

166

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We purified an intracellular pyranose oxidase from mycelial extracts of the white rot fungus Trametes multicolor by using ammonium sulfate fractionation, hydrophobic interaction, ion-exchange chromatography, and gel filtration. The native enzyme has a molecular mass of 270 kDa as determined by equilibrium ultracentrifugation and is composed of four identical 68-kDa subunits as determined by matrix-assisted laser desorption ionization mass spectrometry. Each subunit contains one covalently bound flavin adenine dinucleotide as its prosthetic group. The enzyme oxidizes several aldopyranoses specifically at position C-2, and its preferred electron donor substrates are D-glucose, D-xylose, and L-sorbose. During this oxidation reaction electrons are transferred to oxygen, yielding hydrogen peroxide. In addition, the enzyme catalyzes the twoelectron reduction of 1,4-benzoquinone, several substituted benzoquinones, and 2,6-dichloroindophenol, as well as the one-electron reduction of the ABTS [2,2-azinobis(3-ethylbenzthiazolinesulfonic acid)] cation radical. As judged by the catalytic efficiencies (k cat /K m ), some of these quinone electron acceptors are much better substrates for pyranose oxidase than oxygen. The optimum pH of the pyranose oxidase-catalyzed reaction depends strongly on the electron acceptor employed and varies from 4 to 8. It has been proposed that the main metabolic function of pyranose oxidase is as a constituent of the ligninolytic system of white rot fungi that provides peroxidases with H 2 O 2 . An additional function could be reduction of quinones, key intermediates that are formed during mineralization of lignin.The enzyme pyranose oxidase (P2O) (pyranose:oxygen 2-oxidoreductase; EC 1.1.3.10), which catalyzes the oxidation of several aldopyranoses at position C-2 to yield the corresponding 2-ketoaldoses (aldos-2-uloses, osones), is widely distributed among wood-degrading basidiomycetes (14,32,44). It has been purified and characterized from several microorganisms, including Phanerochaete chrysosporium (2, 45), Phlebiopsis gigantea (39), Pleurotus ostreatus (40), Polyporus obtusus (28), Trametes (Coriolus) versicolor (35), and unidentified basidiomycete no. 52 (26). The currently available data reveal some general similarities among P2Os from these different fungi. Typically, P2O is a rather large, homotetrameric protein that contains covalently bound flavin adenine dinucleotide (FAD). The in vivo substrates of P2O probably are D-glucose, D-galactose, and D-xylose, which are abundant in lignocellulose and which are oxidized to 2-keto-D-glucose (D-arabino-hexos-2-ulose, 2-dehydro-D-glucose), 2-keto-D-galactose (D-lyxo-hexos-2-ulose, 2-dehydro-D-galactose), and 2-keto-D-xylose (D-threopentos-2-ulose, 2-dehydro-D-xylose), respectively. In addition, P2O also exhibits significant activity with a number of other carbohydrates, including L-sorbose, D-glucono-1,5-lactone, and D-allose (20). The substrate selectivity, however, varies to some extent among P2Os isolated from different fungi. During the oxi...

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“…Once these bottlenecks are solved oxidases may become another valuable tool in carbohydrate technology. Goal-oriented screening methods for specific enzymes have recently been developed [27]. A limited overview of some already used or known oxidases will be presented here.…”

Section: Oxidasesmentioning

confidence: 99%

Biotechnical modification of carbohydrates

Vandamme

1995

FEMS Microbiology Reviews

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Goal-oriented screening method for carbohydrate oxidases produced by filamentous fungi

Cited by 26 publications

References 27 publications

Purification and characterization of a pyranose oxidase from the basidiomycete Peniophora gigantea and chemical analyses of its reaction products

Purification and characterization of a pyranose oxidase from the basidiomycete Peniophora gigantea and chemical analyses of its reaction products

Purification and Characterization of Pyranose Oxidase from the White Rot Fungus Trametes multicolor

Biotechnical modification of carbohydrates

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