Production, Characterization, and Reconstitution of Recombinant Quinoprotein Glucose Dehydrogenase (Soluble Type; EC 1.1.99.17) Apoenzyme ofAcinetobacter calcoaceticus

Olsthoorn, Arjen J. J.; Duine, Johannis A.

doi:10.1006/abbi.1996.0530

Cited by 83 publications

(105 citation statements)

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“…The dry material was resuspended in 5 mM Tris-HCl buffer (pH 8.0) and centrifuged at 10,000 ϫ g for 10 min, and the supernatant used as an extract. The pyrrolo-quinoline quinone (PQQ) content was measured enzymatically using the recombinant soluble apoenzyme from Acinetobacter calcoaceticus LMD 79.41 (28). Fifty microliters of the extract and 10 l of 0.4 M CaCl 2 were added to 10 l of apo-glucose dehydrogenase (5 mg/ml).…”

Section: Methodsmentioning

confidence: 99%

Regulation of a Novel Acidithiobacillus caldus Gene Cluster Involved in Metabolism of Reduced Inorganic Sulfur Compounds

Rzhepishevska

Valdés

Marcinkevičienė

et al. 2007

Appl Environ Microbiol

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Acidithiobacillus caldus has been proposed to play a role in the oxidation of reduced inorganic sulfur compounds (RISCs) produced in industrial biomining of sulfidic minerals. Here, we describe the regulation of a new cluster containing the gene encoding tetrathionate hydrolase (tetH), a key enzyme in the RISC metabolism of this bacterium. The cluster contains five cotranscribed genes, ISac1, rsrR, rsrS, tetH, and doxD, coding for a transposase, a two-component response regulator (RsrR and RsrS), tetrathionate hydrolase, and DoxD, respectively. As shown by quantitative PCR, rsrR, tetH, and doxD are upregulated to different degrees in the presence of tetrathionate. Western blot analysis also indicates upregulation of TetH in the presence of tetrathionate, thiosulfate, and pyrite. The tetH cluster is predicted to have two promoters, both of which are functional in Escherichia coli and one of which was mapped by primer extension. A pyrrolo-quinoline quinone binding domain in TetH was predicted by bioinformatic analysis, and the presence of an o-quinone moiety was experimentally verified, suggesting a mechanism for tetrathionate oxidation.

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Section: Methodsmentioning

confidence: 99%

Regulation of a Novel Acidithiobacillus caldus Gene Cluster Involved in Metabolism of Reduced Inorganic Sulfur Compounds

Rzhepishevska

Valdés

Marcinkevičienė

et al. 2007

Appl Environ Microbiol

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“…For M and D, a specific absorption coefficient at 280 nm of 1.28 I g-' cm-' was used. Specific absorption coefficients at 280 nm of 1.67 1 g ~' cin-' for Holo-X and 1 S. 5 I g-cm-for Holo-X,,,, were calculated from the value for Holo,,,, of 1.48 1 g-' cm ' [13] by converting Holo-X into Holo,,,, with glucose and Ca"' . A molar absorption coefficient at 249 nm of 25400 M-' cm-' (at pH 4.0) was used for PQQ 1181.…”

Section: Methodsmentioning

confidence: 99%

“…Determination of Ca" in Holo. Holo-enzyme, reconstituted from D as described [13), was transferred to buffer A via a PDlO and M (---) were calculated from the corresponding peak areas.…”

Section: Methodsmentioning

confidence: 99%

“…the homodimer containing Ca2+ but not PQQ, has been isolated from an Escherichia coli recombinant strain [ 131 (a non-PQQ-producing bacterium). Previous studies [13] on reconstitution suggested that Ca2+ has a dual role in this process and that these can be separately studied. To provide evidence for this, efforts were made to prepare a good-quality monomer (M) preparation (with respect to reconstitutional ability and ab- sence of chelator), to detect species occurring in the pathway of reconstitution, and to study their interconversion.…”

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confidence: 99%

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Ca²⁺ and its Substitutes have Two Different Binding Sites and Roles in Soluble, Quinoprotein (Pyrroloquinoline‐Quinone‐Containing) Glucose Dehydrogenase

Olsthoorn

Otsuki

Duine

1997

European Journal of Biochemistry

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To investigate the mode of binding and the role of Ca2+ in soluble, pyrroloquinoline-quinone (PQQ)-containing glucose dehydrogenase of the bacterium Acinetobacter calcoaceticus (sGDH), the following enzyme species were prepared and their interconversions studied : monomeric apoenzyme (M) ; monomer with one firmly bound Ca2+ ion (M*); dimer consisting of 2 M* (D); dimer consisting of 2 M and 2 PQQ (Holo-Y); dimer consisting of D with 2 PQQ (Holo-X); fully reconstituted enzyme consisting of Holo-X with two extra Ca2+ ions (Holo) or substitutes for Ca2+ (hybrid Holo-enzymes). D and Holo are very stable enzyme species regarding monomerization and inactivation by chelator, respectively, the bound CaZi being locked up in such a way that it is not accessible to chelator. D can be converted into M* by heat treatment and the tightly bound Ca'+ can be removed from M* with chelator, transforming it into M. Reassociation of M* to D occurs spontaneously at 20°C; reassociation of M to D occurs by adding a stoichiometric amount of Ca2+. Synergistic effects were exerted by bound Ca2+ and PQQ, each increasing the affinity of the protein for the other component. Dimerization of M to D occurred with Ca2', Cd2+, Mn2+, and Sr2+ (in decreasing order of effectiveness), but not with Mg", Ba2+, Cozi, Ni", Znz+, or monovalent cations. Conversion of inactive Holo-X into active Holo, was achieved with Ca2+ or metal ions effective in dimerization. Although it is likely that activation of Holo-X involves binding of metal ion to PQQ, the spectral and enzymatic activity differences between normal Holo-and hybrid Holo-enzymes are relatively small. Titration experiments revealed that the two Caz+ ions required for activation of Holo-X are even more firmly bound than the two required for dimerization of M and anchoring of PQQ. Although the two binding sites related with the dual function of Ca2+ show similar metal ion specificity, they are not identical. The presence of two different sites in sGDH appears to be unique because in other PQQ-containing dehydrogenases, the PQQ-containing subunit has only one site. Given the broad spectrum of bivalent metal ions effective in reconstituting quinoprotein dehydrogenase apoenzymes to active holoenzymes, but the limited spectrum for an individual enzyme, the specificity is not so much determined by PQQ but by the variable metal-ion-binding sites.Keywords: quinoprotein ; pyrroloquinoline quinone ; glucose dehydrogenase; reconstitution; calcium.The bacterium Acinetobacter calcoaceticus produces two quite different PQQ-containing (quinoprotein) glucose dehydrogenases, the membrane-bound enzyme (mGDH) and the soluble enzyme (sGDH). mGDH occurs in many gram-negative bacteria [l] and glucose oxidation via this enzyme provides the organism with useful energy [2]. The deduced amino acid sequence of this Correspondence to J. A. Duine,

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“…When PQQ supplemented orally in nano-amounts alleviates the mitogens of B and T cells [118]. PQQ treatment enhances IgA level, restores mass of GALT [119] and induce immunity against bacterial or viral invasion [120], subsequent suppression by increased levels of proteins induced by IFN-β via iNOS, JAK1 and STAT1 signaling pathways (Table 7). PQQ was involved in phosphorylation of 1KKβ, p38 and nF-kB as a pre-inflammatory retort of macrophages [121] increase the number of lymphocytes and CD8+ cells (Table 3) [19].…”

Section: Pqq Role In Immunitymentioning

confidence: 99%

The Life History of Pyrroloquinoline Quinone (PQQ): A Versatile Molecule with Novel Impacts on Living Systems

Naveed¹

2016

IJMBOA

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Pyrroloquinoline quinone (PQQ) acting as redox cofactor of Glucose dehydrogenase is orthocyclic antioxidant acting under multifarious environmental stress and rich conditions in prokaryotes as well as eukaryotes. Microbes are the exclusive source of PQQ biosynthesis and for biocatalysis of glucose into gluconic acid and 2-ketogluconic acid by gram positive and negative bacteria. This study focus to describe PQQ biography highlighted its applications acts as biocatalyst by enhanced production of NADH from pqqC, involved in several important mechanisms like bacterial energy transduction by m-ATPase, production of biocontrol substances, growth stimulating activities and DNA repair. PQQ acting as anti-neurological, anti-degenerative, anti-melanogenic and anti-cancer agent due to its antioxidant nature by scavenging free radicals. PQQ is modulator of immunity by CD4 cells count and IL-2, sleep maintenance by PGC-1 alpha pathway and inflammation due to ROS. The mineral phosphate solubilization results in plant growth promotional, biocontrol, antifungal and ISR activities. PQQ nanoparticles used for production of Biofuels cells and sulfonated polymers. It acts as a modulator of diverse signaling pathways like STAT, MAPK, JAK, JNK, P13K/Akt, mTOR, EGFR and Raps for cell proliferation, differentiation, apoptosis, Box translocation and metabolic pathways by phosphorylation of ADP and suppression of reactive oxygen species. It protects from oxidative stress by acting as Alpha AA modulator of lysine metabolism, TrxR1 in selenium metabolism, low density lipoprotein in lipid metabolism, ATP production in Energy, Glucose and carbohydrate metabolism. It has been structurally characterized with bioinformatics tools and future perspectives being molecular modeling and docking for analytical drug design.

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Production, Characterization, and Reconstitution of Recombinant Quinoprotein Glucose Dehydrogenase (Soluble Type; EC 1.1.99.17) Apoenzyme ofAcinetobacter calcoaceticus

Cited by 83 publications

References 0 publications

Regulation of a Novel Acidithiobacillus caldus Gene Cluster Involved in Metabolism of Reduced Inorganic Sulfur Compounds

Regulation of a Novel Acidithiobacillus caldus Gene Cluster Involved in Metabolism of Reduced Inorganic Sulfur Compounds

Ca²⁺ and its Substitutes have Two Different Binding Sites and Roles in Soluble, Quinoprotein (Pyrroloquinoline‐Quinone‐Containing) Glucose Dehydrogenase

The Life History of Pyrroloquinoline Quinone (PQQ): A Versatile Molecule with Novel Impacts on Living Systems

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Production, Characterization, and Reconstitution of Recombinant Quinoprotein Glucose Dehydrogenase (Soluble Type; EC 1.1.99.17) Apoenzyme ofAcinetobacter calcoaceticus

Cited by 83 publications

References 0 publications

Regulation of a Novel Acidithiobacillus caldus Gene Cluster Involved in Metabolism of Reduced Inorganic Sulfur Compounds

Regulation of a Novel Acidithiobacillus caldus Gene Cluster Involved in Metabolism of Reduced Inorganic Sulfur Compounds

Ca2+ and its Substitutes have Two Different Binding Sites and Roles in Soluble, Quinoprotein (Pyrroloquinoline‐Quinone‐Containing) Glucose Dehydrogenase

The Life History of Pyrroloquinoline Quinone (PQQ): A Versatile Molecule with Novel Impacts on Living Systems

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Ca²⁺ and its Substitutes have Two Different Binding Sites and Roles in Soluble, Quinoprotein (Pyrroloquinoline‐Quinone‐Containing) Glucose Dehydrogenase