Characterization of pyranose dehydrogenase from Agaricus meleagris and its application in the C-2 specific conversion of d-galactose

Sygmund, Christoph; Kittl, Roman; Volc, Jindřich; Halada, Petr; Kubátová, Elena; Haltrich, Dietmar; Peterbauer, Clemens K.

doi:10.1016/j.jbiotec.2007.10.013

Cited by 54 publications

(91 citation statements)

References 26 publications

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“…[42] Synthesis of 3-ketosucrose Sucrose was specifically dehydrogenated by using the pyranose dehydrogenase from Agaricus meleagris (generous gift from Dr. Clemens Peterbauer). [43] The reaction was performed in water at 25 8C with 1,4-benzoquinone as the proton acceptor. The total conversion of sucrose into 3-ketosucrose (2) was observed after 24 h. Its chemical structure was confirmed by NMR spectroscopy.…”

Section: Npas Activity Assaymentioning

confidence: 99%

Probing Substrate Promiscuity of Amylosucrase from Neisseria polysaccharea

Daudé¹,

Champion²,

Morel³

et al. 2013

ChemCatChem

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The amylosucrase from Neisseria polysaccharea (NpAS) naturally catalyzes the synthesis of a variety of products from sucrose and shows signs of plasticity of its active site. To explore further this promiscuity, the tolerance of amylosucrase towards different donor and acceptor substrates was investigated. The selection of alternate donor substrates was first made on the basis of preliminary molecular modeling studies. From 11 potential donors harboring selective derivatizations that were experimentally evaluated, only p‐nitrophenyl‐α‐D‐glucopyranoside was used by the wild‐type enzyme, and this underlines the high specificity of the −1 subsite of NpAS for glucosyl donor substrates. The acceptor substrate promiscuity was further explored by screening 20 hydroxylated molecules, including D‐ and L‐monosaccharides as well as polyols. With the exception of one compound, all were successfully glucosylated, and this showcases the tremendous plasticity of the +1 subsite of NpAS, which is responsible for acceptor recognition. The products obtained from the transglucosylation reactions of three selected acceptors were characterized, and they revealed original structures and enzyme enantiopreference, which were more particularly analyzed by in silico docking analyses.

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Section: Npas Activity Assaymentioning

confidence: 99%

Probing Substrate Promiscuity of Amylosucrase from Neisseria polysaccharea

Daudé¹,

Champion²,

Morel³

et al. 2013

ChemCatChem

View full text Add to dashboard Cite

show abstract

“…GOx has also a relative high Michaelis Menten constant (K M = 27 mM), which limits the maximum current density that can be obtained from human physiological glucose solutions containing normally 5 mM. Therefore it is of great interest in EBFC studies to find new sugar oxidising enzymes that can replace GOx [12,14,15].…”

Section: Introductionmentioning

confidence: 99%

“…The native enzyme is a monomeric glycosylated polypeptide and has a molecular mass of 66,547 Da, containing 7% carbohydrates and one covalently bound flavin adenine dinucleotide (FAD) molecule acting as the redox cofactor [15]. The enzyme exhibits a broad substrate tolerance.…”

Section: Introductionmentioning

confidence: 99%

Wiring of pyranose dehydrogenase with osmium polymers of different redox potentials

Zafar

Tasca

Boland

et al. 2010

Bioelectrochemistry

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“…However, the covalent linkage is not required for the reaction. 2 was prepared using the pET/codAmg gene for the wild-type enzyme as template (38) with forward and reverse oligonucleotides as primers in site-directed mutagenesis. The site-directed mutagenesis was performed using the QuikChange kit, following the manufacturer's manual.…”

mentioning

confidence: 99%

Contribution of Flavin Covalent Linkage with Histidine 99 to the Reaction Catalyzed by Choline Oxidase

Quaye¹,

Cowins

Gadda

2009

Journal of Biological Chemistry

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A number of enzymes, including dehydrogenases (1-3), monooxygenases (4 -7), halogenases (8 -11), and oxidases (7, 12, 13), employ flavin cofactors (FAD or FMN) for their catalytic processes. About a tenth of all flavoproteins have been shown to contain a covalently attached cofactor, which may be linked at the C8M position via histidyl, tyrosyl, or cysteinyl side chains or at the C6M position via a cysteinyl side chain (14). Glucooligosaccharide oxidase (15, 16), hexose oxidase (17), and berberine bridge enzyme (18,19) The discovery of quantum mechanical tunneling in enzymatic reactions, in which hydrogen atoms, protons, and hydride ions are transferred, has attracted considerable interest in enzyme studies geared toward understanding the mechanisms underlying the several orders of magnitudes in the rate enhancements of protein-catalyzed reactions compared with non-enzymatic ones. Tunneling mechanisms have been shown in a wide array of cofactor-dependent enzymes, including flavoenzymes. Examples of flavoenzymes in which the tunneling mechanisms have been demonstrated include morphinone reductase (29, 30), pentaerythritol tetranitrate reductase (29), glucose oxidase (31-33), and choline oxidase (34). Mechanistic data on Class 2 dihydroorotate dehydrogenases, also with a flavin cofactor (FMN) covalently linked to the protein moiety (35,36), could only propose a mechanism that is either stepwise or concerted with significant quantum mechanical tunneling for the hydride transfer from C6 and the deprotonation at C5 in the oxidation of dihydroorotate to orotate (37). This leaves choline oxidase as the only characterized enzyme with a covalently attached flavin cofactor (12,38), where the oxidation of its substrate occurs unequivocally by quantum mechanical tunneling.Choline oxidase from Arthrobacter globiformis catalyzes the two-step FAD-dependent oxidation of the primary alcohol substrate choline to glycine betaine with betaine aldehyde, which is predominantly bound to the enzyme and forms a gem-diol species, as intermediate (Scheme 1). Glycine betaine accumulates in the cytoplasm of plants and bacteria as a defensive mechanism against stress conditions, thus making genetic engineering of relevant plants of economic interest (39 -45), and the biosynthetic pathway for the osmolyte is a potential drug target in human microbial infections of clinical interest (46 -48). The first oxidation step catalyzed by choline oxidase involves the transfer of a hydride ion from a deprotonated choline to the protein-bound flavin followed by reaction of the anionic flavin hydroquinone with molecular oxygen to regenerate the oxidized FAD (for a recent review see Ref. 50). The gem-diol choline, i.e. hydrated betaine aldehyde, is the substrate for the second oxidation step (49), suggesting that the reaction may follow a similar mechanism. The isoalloxazine ring of the flavin cofac-

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Characterization of pyranose dehydrogenase from Agaricus meleagris and its application in the C-2 specific conversion of d-galactose

Cited by 54 publications

References 26 publications

Probing Substrate Promiscuity of Amylosucrase from Neisseria polysaccharea

Probing Substrate Promiscuity of Amylosucrase from Neisseria polysaccharea

Wiring of pyranose dehydrogenase with osmium polymers of different redox potentials

Contribution of Flavin Covalent Linkage with Histidine 99 to the Reaction Catalyzed by Choline Oxidase

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