Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Featherston, E.R.; Mattocks, J.A.; Tirsch, J.L.; Cotruvo, J.A.
    Heterologous expression, purification, and characterization of proteins in the lanthanome (2021), Methods Enzymol., 650, 119-157 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.2.10 gene xoxF, recombinant enzyme expression in Methylorubrum extorquens strain AM1 and in Escherichia coli. gene xoxJ, recombinant expression in Escherichia coli strain BL21(DE3) Methylorubrum extorquens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.2.10 additional information
-
 additional information when La-, Ce-, and Nd-metalated XoxFs are assayed with XoxG as electron acceptor, the Vmax values are not significantly different for the three enzymes, but the Km for XoxG increases from 0.0025 mM for La-XoxF to 0.0076 mM for Nd-XoxF Methylorubrum extorquens
1.1.2.10 0.0025
-
 cytochrome cGJ pH 7.0, 22°C, La-XoxF Methylorubrum extorquens
1.1.2.10 0.0076
-
 cytochrome cGJ pH 7.0, 22°C, Nd-XoxF Methylorubrum extorquens

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.1.2.10 periplasm
-
 Methylorubrum extorquens
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.2.10 Fe2+ in heme and bound to XoxG Methylorubrum extorquens
1.1.2.10 La3+
-
 Methylorubrum extorquens
1.1.2.10 additional information lanthanides, especially the lighter and most abundant members (La, Ce, Pr, Nd, Sm, and Eu) of the lanthanide (Ln) series, are essential for catalysis in the most broadly distributed class of pyrroloquinoline quinone (PQQ)-dependent methanol dehydrogenases (MDHs). The number of distinct lanthanides supporting catalysis in vitro and/or in vivo differs from enzyme to enzyme: e.g. La-Nd, La-Sm/Eu, or La-Gd, according to the XoxF clade, in which an enzyme is found. Strain AM1 XoxF1 can be activated in vivo with La, Ce, Pr, and Nd, and poorly or not at all with Sm. The lanthanides are incorporated when they are added individually to the growth media, XoxF expressed in the presence of La, either from endogenous levels or recombinantly in a methylotroph, show roughly stoichiometric La incorporation, Nd incorporation is more variable. By contrast, plasmid-based expression of XoxF in the presence of Nd leads to substoichiometric Nd insertion into XoxF Methylorubrum extorquens
1.1.2.10 Nd3+
-
 Methylorubrum extorquens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.2.10 methanol + 2 cytochrome cGJ Methylorubrum extorquens
-
 formaldehyde + 2 reduced cytochrome cGJ
-
 r
1.1.2.10 methanol + 2 cytochrome cGJ Methylorubrum extorquens NCIMB 9133
-
 formaldehyde + 2 reduced cytochrome cGJ
-
 r
1.1.2.10 methanol + 2 cytochrome cGJ Methylorubrum extorquens DSM 1338
-
 formaldehyde + 2 reduced cytochrome cGJ
-
 r
1.1.2.10 methanol + 2 cytochrome cGJ Methylorubrum extorquens ATCC 14718
-
 formaldehyde + 2 reduced cytochrome cGJ
-
 r
1.1.2.10 methanol + 2 cytochrome cGJ Methylorubrum extorquens JCM 2805
-
 formaldehyde + 2 reduced cytochrome cGJ
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.2.10 Methylorubrum extorquens P16027 AND P14775 AND P14774 subunit 1/MoxF/XoxF, subunit 2/moxI/XoxJ, and cytochrome cL/moxG/XoxG; Methylobacterium extorquens
-
1.1.2.10 Methylorubrum extorquens ATCC 14718 P16027 AND P14775 AND P14774 subunit 1/MoxF/XoxF, subunit 2/moxI/XoxJ, and cytochrome cL/moxG/XoxG; Methylobacterium extorquens
-
1.1.2.10 Methylorubrum extorquens DSM 1338 P16027 AND P14775 AND P14774 subunit 1/MoxF/XoxF, subunit 2/moxI/XoxJ, and cytochrome cL/moxG/XoxG; Methylobacterium extorquens
-
1.1.2.10 Methylorubrum extorquens JCM 2805 P16027 AND P14775 AND P14774 subunit 1/MoxF/XoxF, subunit 2/moxI/XoxJ, and cytochrome cL/moxG/XoxG; Methylobacterium extorquens
-
1.1.2.10 Methylorubrum extorquens NCIMB 9133 P16027 AND P14775 AND P14774 subunit 1/MoxF/XoxF, subunit 2/moxI/XoxJ, and cytochrome cL/moxG/XoxG; Methylobacterium extorquens
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.2.10 recombinant enzyme from Methylorubrum extorquens strain AM1 and from Escherichia coli, periplasmic extraction is easier in Escherichia coli than in Methylorubrum extorquens, by ammonium sulfate fractionation, cation exchange and hydrophobic interaction chromatography, and desalting gel filtration. Recombinant XoxJ from Escherichia coli strain BL21(DE3) periplasm by anion exchange chromatography, gel filtration, hydrophobic interaction chromatography, and ultrafiltration, followed by gel filtration, and cleavage of the signal peptide Methylorubrum extorquens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.2.10 methanol + 2 cytochrome cGJ
-
 Methylorubrum extorquens formaldehyde + 2 reduced cytochrome cGJ
-
 r
1.1.2.10 methanol + 2 cytochrome cGJ
-
 Methylorubrum extorquens NCIMB 9133 formaldehyde + 2 reduced cytochrome cGJ
-
 r
1.1.2.10 methanol + 2 cytochrome cGJ
-
 Methylorubrum extorquens DSM 1338 formaldehyde + 2 reduced cytochrome cGJ
-
 r
1.1.2.10 methanol + 2 cytochrome cGJ
-
 Methylorubrum extorquens ATCC 14718 formaldehyde + 2 reduced cytochrome cGJ
-
 r
1.1.2.10 methanol + 2 cytochrome cGJ
-
 Methylorubrum extorquens JCM 2805 formaldehyde + 2 reduced cytochrome cGJ
-
 r
1.1.2.10 additional information for assay of chromatography fractions during purifications, activity can be determined by reduction of 2,6-dichlorophenolindolphenol (DCPIP) using phenazine ethosulfate (PES) as an electron acceptor. Development of ddifferent assay methods, detailed overview Methylorubrum extorquens ?
-
-
1.1.2.10 additional information for assay of chromatography fractions during purifications, activity can be determined by reduction of 2,6-dichlorophenolindolphenol (DCPIP) using phenazine ethosulfate (PES) as an electron acceptor. Development of ddifferent assay methods, detailed overview Methylorubrum extorquens NCIMB 9133 ?
-
-
1.1.2.10 additional information for assay of chromatography fractions during purifications, activity can be determined by reduction of 2,6-dichlorophenolindolphenol (DCPIP) using phenazine ethosulfate (PES) as an electron acceptor. Development of ddifferent assay methods, detailed overview Methylorubrum extorquens DSM 1338 ?
-
-
1.1.2.10 additional information for assay of chromatography fractions during purifications, activity can be determined by reduction of 2,6-dichlorophenolindolphenol (DCPIP) using phenazine ethosulfate (PES) as an electron acceptor. Development of ddifferent assay methods, detailed overview Methylorubrum extorquens ATCC 14718 ?
-
-
1.1.2.10 additional information for assay of chromatography fractions during purifications, activity can be determined by reduction of 2,6-dichlorophenolindolphenol (DCPIP) using phenazine ethosulfate (PES) as an electron acceptor. Development of ddifferent assay methods, detailed overview Methylorubrum extorquens JCM 2805 ?
-
-

Subunits

EC Number Subunits Comment Organism
1.1.2.10 ? x * 28600, recombinant XoxJ with cleaved signal peptide, SDS-PAGE Methylorubrum extorquens
1.1.2.10 More XoxF is encoded in an operon alongside genes encoding a c-type cytochrome, XoxG, the physiological electron acceptor for XoxF, as well as a periplasmic solute binding protein (SBP) XoxJ Methylorubrum extorquens

Synonyms

EC Number Synonyms Comment Organism
1.1.2.10 methanol dehydrogenase
-
 Methylorubrum extorquens
1.1.2.10 XoxF
-
 Methylorubrum extorquens
1.1.2.10 XoxJ
-
 Methylorubrum extorquens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.2.10 22
-
 assay at room temperature Methylorubrum extorquens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.2.10 7
-
 assay at Methylorubrum extorquens
1.1.2.10 9
-
 dye-linked assay at Methylorubrum extorquens

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.2.10 cytochrome cGJ c-type cytochrome, XoxG, contains a heme-binding pocket. Recombinant expression in Escherichia coli strain BL21(DE3) and purification from periplasm, method overview. When La-, Ce-, and Nd-metalated XoxFs are assayed with XoxG as electron acceptor, the Vmax values are not significantly different for the three enzymes, but the Km for XoxG increases from 0.0025 mM for La-XoxF to 0.0076 mM for Nd-XoxF. Determination of extinction coefficients and redox potential Methylorubrum extorquens
1.1.2.10 heme bound to XoxG Methylorubrum extorquens
1.1.2.10 pyrroloquinoline quinone PQQ, recombinant expression and binding analysis, method overview Methylorubrum extorquens

General Information

EC Number General Information Comment Organism
1.1.2.10 evolution XoxJ, encoded by the core Ln-MDH operon, is a member of the periplasmic (or solute) binding protein (PBP or SBP) family Methylorubrum extorquens
1.1.2.10 additional information XoxF is encoded in an operon alongside genes encoding a c-type cytochrome, XoxG, the physiological electron acceptor for XoxF, as well as a periplasmic solute binding protein (SBP) XoxJ. The crystal structure of XoxJ reveals general architectures similar to classic SBPs, except it exhibits an exceptionally large cavity, putatively for substrate binding, as well as a beta-sheet missing several strands Methylorubrum extorquens