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Literature summary for 1.5.1.47 extracted from

  • Burton, M.; Abanobi, C.; Wang, K.; Ma, Y.; Rasche, M.
    Substrate specificity analysis of dihydrofolate/dihydromethanopterin reductase homologs in methylotrophic alpha-proteobacteria (2018), Front. Microbiol., 9, 2439 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene dmrA, phylogenetic analysis and tree, recombinant expression of N-terminally His6-tagged and C-terminally His4-tagged enzyme in Escherichia coli strain BL21 Methylorubrum extorquens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics Methylorubrum extorquens
0.102
-
dihydrosarcinapterin pH 6.8, 23°C Hyphomicrobium nitrativorans
0.193
-
7,8-dihydromethanopterin pH 5.3, 23°C, recombinant C-terminally His4-tagged enzyme Methylorubrum extorquens
0.195
-
dihydrosarcinapterin pH 5.3, temperature not specified in the publication Methylorubrum extorquens
0.695
-
dihydrosarcinapterin pH 6.8, 23°C Methylobacterium nodulans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
7,8-dihydromethanopterin + NADPH + H+ Methylorubrum extorquens
-
5,6,7,8-tetrahydromethanopterin + NADP+
-
r
7,8-dihydromethanopterin + NADPH + H+ Methylorubrum extorquens NCIMB 9133
-
5,6,7,8-tetrahydromethanopterin + NADP+
-
r
7,8-dihydromethanopterin + NADPH + H+ Methylorubrum extorquens DSM 1338
-
5,6,7,8-tetrahydromethanopterin + NADP+
-
r
7,8-dihydromethanopterin + NADPH + H+ Methylorubrum extorquens ATCC 14718
-
5,6,7,8-tetrahydromethanopterin + NADP+
-
r
7,8-dihydromethanopterin + NADPH + H+ Methylorubrum extorquens JCM 2805
-
5,6,7,8-tetrahydromethanopterin + NADP+
-
r

Organism

Organism UniProt Comment Textmining
Hyphomicrobium nitrativorans
-
-
-
Methylobacterium nodulans B8IHA6
-
-
Methylobacterium nodulans LMG 21967 B8IHA6
-
-
Methylorubrum extorquens C5B2R8
-
-
Methylorubrum extorquens C5B2R8 Methylobacterium extorquens
-
Methylorubrum extorquens ATCC 14718 C5B2R8 Methylobacterium extorquens
-
Methylorubrum extorquens DSM 1338 C5B2R8
-
-
Methylorubrum extorquens DSM 1338 C5B2R8 Methylobacterium extorquens
-
Methylorubrum extorquens JCM 2805 C5B2R8 Methylobacterium extorquens
-
Methylorubrum extorquens NCIMB 9133 C5B2R8 Methylobacterium extorquens
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzymes from Escherichia coli strain BL21 by nickel affinity chromatography Methylorubrum extorquens

Source Tissue

Source Tissue Comment Organism Textmining
additional information the facultative methylotroph Methylobacterium extorquens AM1, growth on single-carbon (C1) substrates involves the use of both tetrahydromethanopterin (H4MPT) and tetrahydrofolate (H4F) Methylorubrum extorquens
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.63
-
substrate dihydrosarcinapterin, pH 6.8, 23°C Methylobacterium nodulans
2.24
-
substrate dihydrosarcinapterin, pH 5.3, temperature not specified in the publication Methylorubrum extorquens
2.82
-
substrate dihydrosarcinapterin, pH 6.8, 23°C Hyphomicrobium nitrativorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens 5,6,7,8-tetrahydromethanopterin + NADP+
-
?
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens 5,6,7,8-tetrahydromethanopterin + NADP+
-
r
7,8-dihydromethanopterin + NADPH + H+
-
Methylobacterium nodulans 5,6,7,8-tetrahydromethanopterin + NADP+
-
?
7,8-dihydromethanopterin + NADPH + H+
-
Hyphomicrobium nitrativorans 5,6,7,8-tetrahydromethanopterin + NADP+
-
?
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens NCIMB 9133 5,6,7,8-tetrahydromethanopterin + NADP+
-
?
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens NCIMB 9133 5,6,7,8-tetrahydromethanopterin + NADP+
-
r
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens DSM 1338 5,6,7,8-tetrahydromethanopterin + NADP+
-
?
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens DSM 1338 5,6,7,8-tetrahydromethanopterin + NADP+
-
r
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens ATCC 14718 5,6,7,8-tetrahydromethanopterin + NADP+
-
?
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens ATCC 14718 5,6,7,8-tetrahydromethanopterin + NADP+
-
r
7,8-dihydromethanopterin + NADPH + H+
-
Methylobacterium nodulans LMG 21967 5,6,7,8-tetrahydromethanopterin + NADP+
-
?
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens JCM 2805 5,6,7,8-tetrahydromethanopterin + NADP+
-
?
7,8-dihydromethanopterin + NADPH + H+
-
Methylorubrum extorquens JCM 2805 5,6,7,8-tetrahydromethanopterin + NADP+
-
r
dihydrosarcinapterin + NADPH + H+
-
Methylorubrum extorquens tetrahydrosarcinapterin + NADP+
-
?
dihydrosarcinapterin + NADPH + H+
-
Methylobacterium nodulans tetrahydrosarcinapterin + NADP+
-
?
dihydrosarcinapterin + NADPH + H+
-
Hyphomicrobium nitrativorans tetrahydrosarcinapterin + NADP+
-
?
dihydrosarcinapterin + NADPH + H+
-
Methylorubrum extorquens NCIMB 9133 tetrahydrosarcinapterin + NADP+
-
?
dihydrosarcinapterin + NADPH + H+
-
Methylorubrum extorquens DSM 1338 tetrahydrosarcinapterin + NADP+
-
?
dihydrosarcinapterin + NADPH + H+
-
Methylorubrum extorquens ATCC 14718 tetrahydrosarcinapterin + NADP+
-
?
dihydrosarcinapterin + NADPH + H+
-
Methylobacterium nodulans LMG 21967 tetrahydrosarcinapterin + NADP+
-
?
dihydrosarcinapterin + NADPH + H+
-
Methylorubrum extorquens JCM 2805 tetrahydrosarcinapterin + NADP+
-
?
additional information no activity with 7,8-dihydrofolate Methylobacterium nodulans ?
-
-
additional information no activity with 7,8-dihydrofolate Hyphomicrobium nitrativorans ?
-
-
additional information no activity with 7,8-dihydrofolate Methylobacterium nodulans LMG 21967 ?
-
-

Subunits

Subunits Comment Organism
? x * 15800, C-terminally His4-tagged enzyme DmrA, SDS-PAGE Methylorubrum extorquens

Synonyms

Synonyms Comment Organism
dihydromethanopterin reductase
-
Methylorubrum extorquens
DmrA
-
Methylorubrum extorquens
Mnod_4941
-
Methylobacterium nodulans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
15 37 broad Methylobacterium nodulans
15 37 broad Hyphomicrobium nitrativorans
23
-
assay at room temperature Methylorubrum extorquens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5
-
7,8-dihydromethanopterin pH 5.3, 23°C, recombinant C-terminally His4-tagged enzyme Methylorubrum extorquens
1.5
-
dihydrosarcinapterin pH 5.3, temperature not specified in the publication Methylorubrum extorquens
1.6
-
dihydrosarcinapterin pH 6.8, 23°C Methylobacterium nodulans
2.9
-
dihydrosarcinapterin pH 6.8, 23°C Hyphomicrobium nitrativorans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.3
-
assay at Methylorubrum extorquens

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Methylorubrum extorquens

General Information

General Information Comment Organism
evolution homology of DmrA to dihydrofolate reductases leads to the proposal that DmrA evolved from an ancestral dihydrofolate reductase following horizontal transfer of tetrahydromethanopterin (H4MPT) biosynthesis genes from anaerobic archaea to aerobic bacteria. Methylobacterium extorquens AM1 contains one dihydromethanopterin reductase (DmrA) and two putative dihydrofolate reductases, DfrA and DfrB (EC 1.5.1.3), that, respectively, share 26% identity (41% similarity) and 34% identity (53% similarity) with DmrA. DmrA shares no sequence homology with the FMN-containing dihydromethanopterin reductase discovered in archaea (DmrX) or related archaeallike flavoproteins (AfpA and DmrB) from beta-proteobacteria. Phylogenetic analysis and tree. Conformational modeling of DmrA and DfrB Methylorubrum extorquens
metabolism dihydromethanopterin reductase (DmrA) catalyzes the final step of tetrahydromethanopterin (H4MPT) biosynthesis. In the pathways of H4MPT and tetrahydrofolate (H4F) biosynthesis, the last step requires the activity of dihydromethanopterin reductase (Dmr) or dihydrofolate reductase (Dfr). Methylobacterium extorquens AM1 contains one dihydromethanopterin reductase (DmrA) and two putative dihydrofolate reductases, DfrA and DfrB, that, respectively, share 26% identity (41% similarity) and 34% identity (53% similarity) with DmrA Methylorubrum extorquens
physiological function methane-producing archaea and methylotrophic bacteria use tetrahydromethanopterin (H4MPT) and/or tetrahydrofolate (H4F) as coenzymes in one-carbon (C1) transfer pathways. Dihydromethanopterin reductase (DmrA) catalyzes the final step of tetrahydromethanopterin (H4MPT) biosynthesis. The facultative methylotroph Methylobacterium extorquens AM1, growth on single-carbon (C1) substrates involves the use of both tetrahydromethanopterin (H4MPT) and tetrahydrofolate (H4F) Methylorubrum extorquens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
7.77
-
7,8-dihydromethanopterin pH 5.3, 23°C, recombinant C-terminally His4-tagged enzyme Methylorubrum extorquens