Nicotinoprotein methanol dehydrogenase enzymes in Gram-positive methylotrophic bacteria

Hektor, Harm J.; Kloosterman, Harm; Dijkhuizen, Lubbert

doi:10.1016/s1381-1177(99)00073-9

Cited by 23 publications

(14 citation statements)

References 38 publications

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“…While the unambiguous presence of this cofactor in an XoxF-type large subunit was established only quite recently (43), the finding was expected. Except for nicotinoprotein methanol/alcohol dehydrogenase (3,4), all of the MDHs known to date rely on PQQ as the catalytic center and PQQ-binding motifs have been identified before in XoxF MDHs (see Fig. S1A) (8,32).…”

Section: Discussionmentioning

confidence: 99%

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XoxF-Type Methanol Dehydrogenase from the Anaerobic Methanotroph “Candidatus Methylomirabilis oxyfera”

Wessels

Pol

et al. 2015

Appl Environ Microbiol

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b "Candidatus Methylomirabilis oxyfera" is a newly discovered anaerobic methanotroph that, surprisingly, oxidizes methane through an aerobic methane oxidation pathway. The second step in this aerobic pathway is the oxidation of methanol. In Gramnegative bacteria, the reaction is catalyzed by pyrroloquinoline quinone (PQQ)-dependent methanol dehydrogenase (MDH). The genome of "Ca. Methylomirabilis oxyfera" putatively encodes three different MDHs that are localized in one large gene cluster: one so-called MxaFI-type MDH and two XoxF-type MDHs (XoxF1 and XoxF2). MxaFI MDHs represent the canonical enzymes, which are composed of two PQQ-containing large (␣) subunits (MxaF) and two small (␤) subunits (MxaI). XoxF MDHs are novel, ecologically widespread, but poorly investigated types of MDHs that can be phylogenetically divided into at least five different clades. The XoxF MDHs described thus far are homodimeric proteins containing a large subunit only. Here, we purified a heterotetrameric MDH from "Ca. Methylomirabilis oxyfera" that consisted of two XoxF and two MxaI subunits. The enzyme was localized in the periplasm of "Ca. Methylomirabilis oxyfera" cells and catalyzed methanol oxidation with appreciable specific activity and affinity (V max of 10 mol min ؊1 mg ؊1 protein, K m of 17 M). PQQ was present as the prosthetic group, which has to be taken up from the environment since the known gene inventory required for the synthesis of this cofactor is lacking. The MDH from "Ca. Methylomirabilis oxyfera" is the first representative of type 1 XoxF proteins to be described.

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Section: Discussionmentioning

confidence: 99%

“…MDHs fall into different classes. Gram-positive microorganisms harbor nicotinoprotein MDHs in their cytoplasm that use NAD(P)H as the electron acceptor for methanol oxidation (3,4). In Gram-negative bacteria, a phylogenetically unrelated protein is present in the periplasm that possesses pyrroloquinoline quinone (PQQ) as the catalytic center (5-7).…”

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confidence: 99%

XoxF-Type Methanol Dehydrogenase from the Anaerobic Methanotroph “Candidatus Methylomirabilis oxyfera”

Wessels

Pol

et al. 2015

Appl Environ Microbiol

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“…Nicotino-enzymes are not only involved in oxidation of alcohols (18), but can also perform redox-silent reactions such as epimerizations (e.g. UDP-galactose epimerase) and more complex reactions (e.g.…”

Section: Discussionmentioning

confidence: 99%

Multiple Turnovers of the Nicotino-Enzyme PdxB Require α-Keto Acids as Cosubstrates

2010

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PdxB catalyzes the second step in the biosynthesis of pyridoxal phosphate by oxidizing 4-phospho-D-erythronate (4PE) to 2-oxo-3-hydroxy-4-phospho-butanoate (OHPB) with concomitant reduction of NAD+ to NADH. PdxB is a nicotino-enzyme wherein the NAD(H) cofactor remains tightly bound to PdxB. It has been a mystery how PdxB performs multiple turnovers since addition of free NAD+ does not re-oxidize the enzyme-bound NADH following conversion of 4PE to OHPB. We have solved this mystery by demonstrating that a variety of physiologically available α-ketoacids serve as oxidants of PdxB to sustain multiple turnovers. In a coupled assay using the next two enzymes of the biosynthetic pathway for pyridoxal phosphate (SerC and PdxA), we have found that α-ketoglutarate, oxaloacetic acid, and pyruvate are equally good substrates for PdxB (kcat/Km values ~ 1 × 104 M-1s-1). The kinetic parameters for the substrate 4PE include a kcat of 1.4 s-1, a Km of 2.9 μM, and a kcat/Km of 6.7 × 106 M-1s-1. Additionally, we have characterized the stereochemistry of α-ketoglutarate reduction by showing that D-2-HGA, but not L-2-HGA, is a competitive inhibitor vs. 4PE and a noncompetitive inhibitor vs. α-ketoglutarate.

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“…This MDO is similar to the NAD-dependent MDH of B. methanolicus C1 in quaternary structure, cofactor composition, tightly but non-covalently bound NAD(P)(H) and enzymic properties (Bystrykh et al, 1993b;Hektor et al, 2002;van Ophem et al, 1993;Vonck et al, 1991). The MDO also has enzymic properties different from those of the B. methanolicus C1 MDH: MDO exhibits DMNA-dependent MDH, NADHdependent formaldehyde reductase and formaldehyde dismutase activities (Hektor et al, 2000), while B. methanolicus C1 MDH only displays NAD-dependent MDH and NADH-dependent formaldehyde reductase activities (Arfman et al, 1989). The Gram-positive bacterium Rhodococcus sp.…”

Section: Introductionmentioning

confidence: 99%

Identification and functional characterization of a gene for the methanol : N,N'-dimethyl-4-nitrosoaniline oxidoreductase from Mycobacterium sp. strain JC1 (DSM 3803)

Park

Lee

et al. 2009

Microbiology

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Mycobacterium sp. strain JC1 is able to grow on methanol as a sole source of carbon and energy using methanol : N,N′-dimethyl-4-nitrosoaniline oxidoreductase (MDO) as a key enzyme for primary methanol oxidation. Purified MDO oxidizes ethanol and formaldehyde as well as methanol. The Mycobacterium sp. strain JC1 gene for MDO (mdo) was cloned, sequenced, and determined to have an open reading frame of 1272 bp. Northern blot and promoter analysis revealed that mdo transcription was induced in cells grown in the presence of methanol. Northern blotting together with RT-PCR also showed that the mdo gene was transcribed as monocistronic mRNA. Primer extension analysis revealed that the transcriptional start site of the mdo gene is located 21 bp upstream of the mdo start codon. An mdo-deficient mutant of Mycobacterium sp. strain JC1 did not grow with methanol as a sole source of carbon and energy.

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Nicotinoprotein methanol dehydrogenase enzymes in Gram-positive methylotrophic bacteria

Cited by 23 publications

References 38 publications

XoxF-Type Methanol Dehydrogenase from the Anaerobic Methanotroph “Candidatus Methylomirabilis oxyfera”

XoxF-Type Methanol Dehydrogenase from the Anaerobic Methanotroph “Candidatus Methylomirabilis oxyfera”

Multiple Turnovers of the Nicotino-Enzyme PdxB Require α-Keto Acids as Cosubstrates

Identification and functional characterization of a gene for the methanol : N,N'-dimethyl-4-nitrosoaniline oxidoreductase from Mycobacterium sp. strain JC1 (DSM 3803)

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