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Literature summary for 1.1.1.244 extracted from

  • Mueller, J.E.; Meyer, F.; Litsanov, B.; Kiefer, P.; Potthoff, E.; Heux, S.; Quax, W.J.; Wendisch, V.F.; Brautaset, T.; Portais, J.C.; Vorholt, J.A.
    Engineering Escherichia coli for methanol conversion (2015), Metab. Eng., 28, 190-201 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
Act activator protein, UniProt ID I3EA59, the in vitro activity of Bacillus coagulans Adh is increased by the endogenous activator protein Act, a Nudix hydrolase Weizmannia coagulans
Act activator protein, UniProt ID I3EA59, the in vitro activity of Bacillus methanolicus Mdh is increased by the endogenous activator protein Act, a Nudix hydrolase, no activation in vivo Bacillus methanolicus
Act activator protein, UniProt ID I3EA59, the in vitro activity of Bacillus methanolicus Mdh3 is increased by the endogenous activator protein Act, a Nudix hydrolase Bacillus methanolicus
Act activator protein, UniProt ID I3EA59, the in vitro activity of Desulfitobacterium hafniense Adh is increased by the endogenous activator protein Act, a Nudix hydrolase Desulfitobacterium hafniense
Act protein activator protein, UniProt ID I3EA59, the in vitro activity of Lysinibacillus sphaericus Mdh is increased by the endogenous activator protein Act, a Nudix hydrolase Lysinibacillus sphaericus
additional information no activation by activator protein Act, UniProt ID I3EA59 Lysinibacillus fusiformis

Cloned(Commentary)

Cloned (Comment) Organism
gene BFZC1_05383, recombinant expression in Escherichia coli Lysinibacillus fusiformis
gene Bsph_4187, recombinant expression in Escherichia coli Lysinibacillus sphaericus
gene mdh, recombinant expression in Escherichia coli Bacillus methanolicus
gene mdh1, recombinant expression in Escherichia coli Bacillus methanolicus
gene mdh2, recombinant expression in Escherichia coli Bacillus methanolicus
gene mdh3, recombinant expression in Escherichia coli Bacillus methanolicus
recombinant enzyme expression in Escherichia coli Weizmannia coagulans
recombinant enzyme expression in Escherichia coli Desulfitobacterium hafniense
recombinant enzyme expression in Escherichia coli Desulfofundulus kuznetsovii

Protein Variants

Protein Variants Comment Organism
D38G site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme Bacillus methanolicus
D41G site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Bacillus methanolicus
additional information utilization of methanol in Escherichia coli is allowed by the implementation of NAD-dependent methanol dehydrogenase and the establishment of the ribulose monophosphate cycle by expressing the genes for hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), system evaluation, overview Weizmannia coagulans
additional information utilization of methanol in Escherichia coli is allowed by the implementation of NAD-dependent methanol dehydrogenase and the establishment of the ribulose monophosphate cycle by expressing the genes for hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), system evaluation, overview Desulfitobacterium hafniense
additional information utilization of methanol in Escherichia coli is allowed by the implementation of NAD-dependent methanol dehydrogenase and the establishment of the ribulose monophosphate cycle by expressing the genes for hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), system evaluation, overview Lysinibacillus fusiformis
additional information utilization of methanol in Escherichia coli is allowed by the implementation of NAD-dependent methanol dehydrogenase and the establishment of the ribulose monophosphate cycle by expressing the genes for hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), system evaluation, overview Bacillus methanolicus
additional information utilization of methanol in Escherichia coli is allowed by the implementation of NAD-dependent methanol dehydrogenase and the establishment of the ribulose monophosphate cycle by expressing the genes for hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), system evaluation, overview Lysinibacillus sphaericus
additional information utilization of methanol in Escherichia coli is allowed by the implementation of NAD-dependent methanol dehydrogenase and the establishment of the ribulose monophosphate cycle by expressing the genes for hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), system evaluation, overview Desulfofundulus kuznetsovii
additional information utilization of methanol in Escherichia coli is allowed by the implementation of NAD-dependent methanol dehydrogenase and the establishment of the ribulose monophosphate cycle by expressing the genes for hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), up to 40% incorporation of methanol into central metabolites is achieved, system evaluation, overview Bacillus methanolicus
S97G site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme Bacillus methanolicus
S97G site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Bacillus methanolicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
methanol + NAD+ Weizmannia coagulans
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Desulfitobacterium hafniense
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Lysinibacillus fusiformis
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Bacillus methanolicus
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Lysinibacillus sphaericus
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Desulfofundulus kuznetsovii
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Desulfofundulus kuznetsovii MGA3
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Lysinibacillus fusiformis ZC1
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Desulfitobacterium hafniense Y51
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Bacillus methanolicus PB1
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Weizmannia coagulans 36D1
-
formaldehyde + NADH + H+
-
r
methanol + NAD+ Bacillus methanolicus MGA3
-
formaldehyde + NADH + H+
-
r

Organism

Organism UniProt Comment Textmining
Bacillus methanolicus I3DTM5
-
-
Bacillus methanolicus I3DTP7
-
-
Bacillus methanolicus I3DVX6
-
-
Bacillus methanolicus I3DX19
-
-
Bacillus methanolicus I3E2P9
-
-
Bacillus methanolicus I3E949
-
-
Bacillus methanolicus MGA3 I3DTM5
-
-
Bacillus methanolicus MGA3 I3E2P9
-
-
Bacillus methanolicus MGA3 I3E949
-
-
Bacillus methanolicus PB1 I3DTP7
-
-
Bacillus methanolicus PB1 I3DVX6
-
-
Bacillus methanolicus PB1 I3DX19
-
-
Desulfitobacterium hafniense
-
-
-
Desulfitobacterium hafniense Y51
-
-
-
Desulfofundulus kuznetsovii
-
-
-
Desulfofundulus kuznetsovii MGA3
-
-
-
Lysinibacillus fusiformis
-
-
-
Lysinibacillus fusiformis ZC1
-
-
-
Lysinibacillus sphaericus B1HX72
-
-
Weizmannia coagulans
-
-
-
Weizmannia coagulans 36D1
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
additional information
-
no activity of the recombinant enzyme in Escherichia coli suspension cells Lysinibacillus fusiformis
additional information
-
no activity of the recombinant enzyme in Escherichia coli suspension cells Bacillus methanolicus
0.0007
-
recombinant wild-type Mdh in Escherichia coli suspension cells, pH 7.4, 37┬░C, with addition of Act Bacillus methanolicus
0.0007
-
recombinant wild-type Mdh2 in Escherichia coli suspension cells, pH 7.4, 37┬░C Bacillus methanolicus
0.001
-
recombinant wild-type Adh in Escherichia coli suspension cells, pH 7.4, 37┬░C Lysinibacillus sphaericus
0.0014
-
recombinant wild-type Mdh in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Bacillus methanolicus
0.0015
-
recombinant wild-type Mdh3 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Bacillus methanolicus
0.0017
-
recombinant wild-type Mdh in Escherichia coli suspension cells, pH 7.4, 37┬░C Bacillus methanolicus
0.0018
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Desulfitobacterium hafniense
0.0023
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Bacillus methanolicus
0.0026
-
recombinant wild-type Mdh in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Bacillus methanolicus
0.0029
-
recombinant wild-type Adh in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Lysinibacillus sphaericus
0.003
-
recombinant wild-type Mdh2 in Escherichia coli suspension cells, pH 7.4, 37┬░C Weizmannia coagulans
0.003
-
recombinant wild-type Mdh2 in Escherichia coli suspension cells, pH 7.4, 37┬░C Bacillus methanolicus
0.0033
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Bacillus methanolicus
0.0038
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Lysinibacillus fusiformis
0.0038
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C, with addition of Act Lysinibacillus fusiformis
0.0051
-
recombinant mutant S97G Mdh in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Bacillus methanolicus
0.0057
-
recombinant mutant S97G Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Bacillus methanolicus
0.0058
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Weizmannia coagulans
0.007
-
recombinant wild-type Mdh2 in Escherichia coli suspension cells, pH 7.4, 37┬░C Desulfitobacterium hafniense
0.008 4 recombinant wild-type Adh in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C, with addition of Act Lysinibacillus sphaericus
0.01
-
recombinant wild-type Mdh1 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C Bacillus methanolicus
0.0114
-
recombinant mutant S97G Mdh2 in Escherichia coli suspension cells, pH 7.4, 37┬░C Bacillus methanolicus
0.014
-
recombinant mutant S97G Mdh in Escherichia coli suspension cells, pH 7.4, 37┬░C Bacillus methanolicus
0.0155
-
recombinant wild-type Mdh in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C, with addition of Act Bacillus methanolicus
0.0155
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C, with addition of Act Weizmannia coagulans
0.0167
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C, with addition of Act Desulfitobacterium hafniense
0.021
-
recombinant mutant S97G Mdh in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C, with addition of Act Bacillus methanolicus
0.0213
-
recombinant wild-type Mdh2 in Escherichia coli suspension cells, pH 7.4, 37┬░C, with addition of Act Bacillus methanolicus
0.023
-
recombinant wild-type Mdh3 in Escherichia coli suspension cells, pH 7.4, 37┬░C Bacillus methanolicus
0.0273
-
recombinant wild-type Mdh3 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C, with addition of Act Bacillus methanolicus
0.03
-
recombinant wild-type Mdh2 in Escherichia coli suspension cells, pH 7.4, 37┬░C Bacillus methanolicus
0.045
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 7.4, 37┬░C, with addition of Act Bacillus methanolicus
0.22
-
recombinant wild-type Mdh2 in Escherichia coli cell-free enzyme extract, pH 9.5, 45┬░C Bacillus methanolicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
methanol + NAD+
-
Weizmannia coagulans formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Desulfitobacterium hafniense formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Lysinibacillus fusiformis formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Bacillus methanolicus formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Lysinibacillus sphaericus formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Desulfofundulus kuznetsovii formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Desulfofundulus kuznetsovii MGA3 formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Lysinibacillus fusiformis ZC1 formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Desulfitobacterium hafniense Y51 formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Bacillus methanolicus PB1 formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Weizmannia coagulans 36D1 formaldehyde + NADH + H+
-
r
methanol + NAD+
-
Bacillus methanolicus MGA3 formaldehyde + NADH + H+
-
r

Synonyms

Synonyms Comment Organism
ADH
-
Weizmannia coagulans
ADH
-
Desulfitobacterium hafniense
ADH
-
Lysinibacillus fusiformis
ADH
-
Lysinibacillus sphaericus
ADH
-
Desulfofundulus kuznetsovii
BFZC1_05383
-
Lysinibacillus fusiformis
Bsph_4187
-
Lysinibacillus sphaericus
MDH
-
Bacillus methanolicus
Mdh1
-
Bacillus methanolicus
MDH2
-
Bacillus methanolicus
Mdh3
-
Bacillus methanolicus
NAD-dependent methanol dehydrogenase
-
Weizmannia coagulans
NAD-dependent methanol dehydrogenase
-
Desulfitobacterium hafniense
NAD-dependent methanol dehydrogenase
-
Lysinibacillus fusiformis
NAD-dependent methanol dehydrogenase
-
Bacillus methanolicus
NAD-dependent methanol dehydrogenase
-
Lysinibacillus sphaericus
NAD-dependent methanol dehydrogenase
-
Desulfofundulus kuznetsovii

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
37
-
assay at Weizmannia coagulans
37
-
assay at Desulfitobacterium hafniense
37
-
assay at Lysinibacillus fusiformis
37
-
assay at Bacillus methanolicus
37
-
assay at Lysinibacillus sphaericus
37
-
assay at Desulfofundulus kuznetsovii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Weizmannia coagulans
7.4
-
assay at Desulfitobacterium hafniense
7.4
-
assay at Lysinibacillus fusiformis
7.4
-
assay at Bacillus methanolicus
7.4
-
assay at Lysinibacillus sphaericus
7.4
-
assay at Desulfofundulus kuznetsovii

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Weizmannia coagulans
NAD+
-
Desulfitobacterium hafniense
NAD+
-
Lysinibacillus fusiformis
NAD+
-
Bacillus methanolicus
NAD+
-
Lysinibacillus sphaericus
NAD+
-
Desulfofundulus kuznetsovii