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

BRENDA support

Literature summary for 1.1.2.10 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)

Cloned (Comment) Organism
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
0.0025
-
cytochrome cGJ pH 7.0, 22┬░C, La-XoxF Methylorubrum extorquens
0.0076
-
cytochrome cGJ pH 7.0, 22┬░C, Nd-XoxF Methylorubrum extorquens

Localization

Localization Comment Organism GeneOntology No. Textmining
periplasm
-
Methylorubrum extorquens
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ in heme and bound to XoxG Methylorubrum extorquens
La3+
-
Methylorubrum extorquens
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
Nd3+
-
Methylorubrum extorquens

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Comment) Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
methanol + 2 cytochrome cGJ
-
Methylorubrum extorquens formaldehyde + 2 reduced cytochrome cGJ
-
r
methanol + 2 cytochrome cGJ
-
Methylorubrum extorquens NCIMB 9133 formaldehyde + 2 reduced cytochrome cGJ
-
r
methanol + 2 cytochrome cGJ
-
Methylorubrum extorquens DSM 1338 formaldehyde + 2 reduced cytochrome cGJ
-
r
methanol + 2 cytochrome cGJ
-
Methylorubrum extorquens ATCC 14718 formaldehyde + 2 reduced cytochrome cGJ
-
r
methanol + 2 cytochrome cGJ
-
Methylorubrum extorquens JCM 2805 formaldehyde + 2 reduced cytochrome cGJ
-
r
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 ?
-
-
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 ?
-
-
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 ?
-
-
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 ?
-
-
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

Subunits Comment Organism
? x * 28600, recombinant XoxJ with cleaved signal peptide, SDS-PAGE Methylorubrum extorquens
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

Synonyms Comment Organism
methanol dehydrogenase
-
Methylorubrum extorquens
XoxF
-
Methylorubrum extorquens
XoxJ
-
Methylorubrum extorquens

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
22
-
assay at room temperature Methylorubrum extorquens

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
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
heme bound to XoxG Methylorubrum extorquens
pyrroloquinoline quinone PQQ, recombinant expression and binding analysis, method overview Methylorubrum extorquens

General Information

General Information Comment Organism
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
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