Catalytic properties and crystal structure of quinoprotein aldose sugar dehydrogenase from hyperthermophilic archaeon Pyrobaculum aerophilum

Sakuraba, Haruhiko; Yokono, K.; Yoneda, Kazunari; Watanabe, Akira; Asada, Yasuhiko; Satomura, Takenori; Yabutani, Tomoki; Motonaka, Junko; Ohshima, Toshihisa

doi:10.1016/j.abb.2010.08.002

Cited by 32 publications

(35 citation statements)

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“…PQQ-dependent quinoproteins are divided into the following two groups based on their structure: those containing an eight-bladed ␤-propeller where each blade consists of a four-stranded antiparallel ␤-sheet (29)(30)(31)(32) and those with a six-bladed ␤-propeller, such as soluble glucose dehydrogenase from A. calcoaceticus and soluble Asd from E. coli and P. aerophilum (27,33,34). Members of these two groups do not have significant primary sequence homology.…”

Section: Discussionmentioning

confidence: 99%

“…The black arrowhead indicates the predicted catalytic histidine residue. Open arrowheads indicate conserved residues involved in PQQ binding in Pa2KGDH and known quinoproteins (27,33,34 For phylogenetic analysis of the bacterial PQQ-dependent enzymes, complete amino acid sequences were initially aligned using MAFFT and then manually edited using SeaView (22). A phylogenetic tree was constructed from this alignment by using the neighbor-joining method (23) in ClustalX software (24), with 1,000 bootstraps.…”

Section: Methodsmentioning

confidence: 99%

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A Novel Pyrroloquinoline Quinone-Dependent 2-Keto- d -Glucose Dehydrogenase from Pseudomonas aureofaciens

et al. 2015

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A gene encoding an enzyme similar to a pyrroloquinoline quinone (PQQ)-dependent sugar dehydrogenase from filamentous fungi, which belongs to new auxiliary activities (AA) family 12 in the CAZy database, was cloned from Pseudomonas aureofaciens. The deduced amino acid sequence of the cloned enzyme showed only low homology to previously characterized PQQ-dependent enzymes, and multiple-sequence alignment analysis showed that the enzyme lacks one of the three conserved arginine residues that function as PQQ-binding residues in known PQQ-dependent enzymes. The recombinant enzyme was heterologously expressed in an Escherichia coli expression system for further characterization. The UV-visible (UV-Vis) absorption spectrum of the oxidized form of the holoenzyme, prepared by incubating the apoenzyme with PQQ and CaCl 2 , revealed a broad peak at approximately 350 nm, indicating that the enzyme binds PQQ. With the addition of 2-keto-D-glucose (2KG) to the holoenzyme solution, a sharp peak appeared at 331 nm, attributed to the reduction of PQQ bound to the enzyme, whereas no effect was observed upon 2KG addition to authentic PQQ. Enzymatic assay showed that the recombinant enzyme specifically reacted with 2KG in the presence of an appropriate electron acceptor, such as 2,6-dichlorophenol indophenol, when PQQ and CaCl 2 were added. 1 H nuclear magnetic resonance ( 1 H-NMR) analysis of reaction products revealed 2-keto-D-gluconic acid (2KGA) as the main product, clearly indicating that the recombinant enzyme oxidizes the C-1 position of 2KG. Therefore, the enzyme was identified as a PQQ-dependent 2KG dehydrogenase (Pa2KGDH). Considering the high substrate specificity, the physiological function of Pa2KGDH may be for production of 2KGA. P yrroloquinoline quinone (PQQ) is a major cofactor in redox enzymes called quinoproteins and was first identified as a cofactor in bacterial methanol dehydrogenase (1) and glucose dehydrogenase (2), in 1979. The presence of PQQ is a defining feature of quinoprotein enzymes, which distinguishes them from nicotinamide-and flavin-dependent enzymes. In nature, PQQ-dependent quinoproteins have primarily been found as bacterial proteins, localized to the periplasm or bound to membranes, which catalyze the oxidation of various sugars and alcohols, such as glucose, methanol, and ethanol, in the presence of an appropriate electron acceptor (3). Because bacterial PQQ-dependent enzymes require cytochrome c or ubiquinone as an electron acceptor, they are believed to be involved in cellular respiration (4, 5).Several Gram-negative bacteria, such as Pseudomonas spp. and Gluconobacter spp., possess a pathway for the oxidation of monosaccharides, known as oxidative fermentation. Among these bacteria, Pseudomonas spp. have been reported to accumulate an oxidized form of glucose in culture (6), producing 2-keto-D-gluconic acid (2KGA) from D-glucose, with D-gluconic acid produced as a metabolic intermediate. The 2KGA biosynthetic pathway proceeds through the sequential catalytic actions of two membrane-bound d...

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

A Novel Pyrroloquinoline Quinone-Dependent 2-Keto- d -Glucose Dehydrogenase from Pseudomonas aureofaciens

et al. 2015

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Section: Introductionmentioning

confidence: 99%

“…21 Additionally, this enzyme shows outstanding thermal stability; activity remained almost at the same level even after incubation at 100 C for 10 min. 21 The tPQQ-ASD is one of the most thermostable enzymes among PQQ-GDHs and PQQ-ASDs, which are reported to show high thermal stability. [8][9][10][11] With respect to the practical use of enzyme sensors and biofuel cells one of the most important factors is the stable immobilization of all components related to the electron transfer process, from the substrates to the electrode.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, a novel thermophilic PQQ-ASD (tPQQ-ASD) derived from hyperthermophilic archaeon Pyrobaculum aerophilum was reported by Sakuraba et al 21 The tPQQ-ASD works on a broad range of aldose sugars, and it has high Km values and low Kcat values to the substrates. 21 Additionally, this enzyme shows outstanding thermal stability; activity remained almost at the same level even after incubation at 100 C for 10 min.…”

Section: Introductionmentioning

confidence: 99%

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Fabrication and Characterization of a Thermostable Quinoprotein Aldose Sugar Dehydrogenase Immobilized Electrode

et al. 2013

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We fabricated a thermostable quinoprotein aldose sugar dehydrogenase (tPQQ-ASD derived from a hyperthermophilic archaeon Pyrobaculum aerophilum) immobilized electrode. The electrode was prepared by immobilizing agarose gel mixed with the enzyme and carbon nanofiber (CNF) on a carbon paste (CP) electrode containing p-benzoquinone (BQ) as an electron mediator. The electrocatalytic response was clearly observed by the addition of D-glucose at the electrode. The electrode properties such as pH, temperature dependency and substrate selectivity basically followed the enzyme properties. The current response against D-glucose increased with measurement temperatures up to 70 C, and response perturbation caused by dissolved oxygen level was not observed at the electrode. As for the results of long-term stability evaluation, the current response was stable for 30 days when the electrode was stored in HEPES buffer solution (pH 7.0) at 4 C.

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Pyrroloquinoline Quinone ( PQQ )

Toyama

2016

Industrial Biotechnology of Vitamins, Biopigments, and Antioxidants

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Catalytic properties and crystal structure of quinoprotein aldose sugar dehydrogenase from hyperthermophilic archaeon Pyrobaculum aerophilum

Cited by 32 publications

References 30 publications

A Novel Pyrroloquinoline Quinone-Dependent 2-Keto- d -Glucose Dehydrogenase from Pseudomonas aureofaciens

A Novel Pyrroloquinoline Quinone-Dependent 2-Keto- d -Glucose Dehydrogenase from Pseudomonas aureofaciens

Fabrication and Characterization of a Thermostable Quinoprotein Aldose Sugar Dehydrogenase Immobilized Electrode

Pyrroloquinoline Quinone ( PQQ )

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