Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | FrmB is induced 45-75fold by formaldehyde, while YeiG is constitutively expressed | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
genes frmB or yaiM, and yeiG, subcloning in strain DH5alpha, overexpression in strain BL21(DE3) of His-tagged wild-type and mutant enzymes | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
C26A | site-directed mutagenesis, the YeiG mutant shows wild-type activity | Escherichia coli |
C54A | site-directed mutagenesis, the YeiG mutant shows wild-type activity | Escherichia coli |
D199A | site-directed mutagenesis, the YeiG mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
D218A | site-directed mutagenesis, the YeiG mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
D223A | site-directed mutagenesis, the YeiG mutant shows no esterase activity | Escherichia coli |
D255A | site-directed mutagenesis, the YeiG mutant shows 2fold reduced activity compared to the wild-type enzyme | Escherichia coli |
D80A | site-directed mutagenesis, the YeiG mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
additional information | construction of FrmB and YeiG deletion mutants | Escherichia coli |
S154A | site-directed mutagenesis, the YeiG mutant shows no esterase activity | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
NEM | inhibition of wild-type YeiG and YeiG mutant C26A, no inhibition of YeiG mutant C54A | Escherichia coli | |
PMSF | inhibition of wild-type YeiG and YeiG mutants C54A and C26A | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.29 | - |
4-nitrophenyl acetate | pH 8.0, 37°C, recombinant FrmB | Escherichia coli | |
0.41 | - |
S-formylglutathione | pH 8.0, 37°C, recombinant FrmB | Escherichia coli | |
0.43 | - |
S-formylglutathione | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
0.45 | - |
4-nitrophenyl acetate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
0.48 | - |
4-nitrophenyl propionate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
0.54 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant D255A | Escherichia coli | |
0.58 | - |
S-Lactoylglutathione | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
0.6 | - |
S-Lactoylglutathione | pH 8.0, 37°C, recombinant FrmB | Escherichia coli | |
0.7 | - |
4-nitrophenyl butyrate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
0.8 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant D80A | Escherichia coli | |
0.83 | - |
4-nitrophenyl propionate | pH 8.0, 37°C, recombinant FrmB | Escherichia coli | |
0.95 | - |
4-nitrophenyl caproate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
1.01 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant D199A | Escherichia coli | |
1.03 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
1.06 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant C26A | Escherichia coli | |
1.28 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant C54A | Escherichia coli | |
1.3 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant D218A | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-formylglutathione + H2O | Escherichia coli | the substrate is an intermediate in the glutathione-dependent formaldehyde detoxification pathway | glutathione + formate | - |
? | |
S-formylglutathione + H2O | Escherichia coli BW25113 | the substrate is an intermediate in the glutathione-dependent formaldehyde detoxification pathway | glutathione + formate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
2 enzymes FrmB and YeiG, encoded by genes frmB or yaiM, and yeiG | - |
Escherichia coli BW25113 | - |
2 enzymes FrmB and YeiG, encoded by genes frmB or yaiM, and yeiG | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from strain BL21(DE3) by nickel affinity chromatography | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
S-formylglutathione + H2O = glutathione + formate | Ser145, Asp233, and His256 are essential for catalysis, representing a serine hydrolase catalytic triad, Cys26 and Cys54 are not involved in catalysis, mechanism | Escherichia coli |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | expression levels of wild-type and mutant enzymes, induced or uninduced, overview | Escherichia coli | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
substrate specificity | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-naphthyl acetate + H2O | artificial substrate of YeiG | Escherichia coli | 2-naphthol + acetate | - |
? | |
2-naphthyl acetate + H2O | artificial substrate of YeiG | Escherichia coli BW25113 | 2-naphthol + acetate | - |
? | |
4-nitrophenyl acetate + H2O | artificial substrate of FrmB and YeiG | Escherichia coli | 4-nitrophenol + acetate | - |
? | |
4-nitrophenyl acetate + H2O | artificial substrate of FrmB and YeiG | Escherichia coli BW25113 | 4-nitrophenol + acetate | - |
? | |
4-nitrophenyl butyrate + H2O | artificial substrate of YeiG | Escherichia coli | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl caproate + H2O | artificial substrate of YeiG | Escherichia coli | 4-nitrophenol + caproate | - |
? | |
4-nitrophenyl propionate + H2O | artificial substrate of FrmB and YeiG | Escherichia coli | 4-nitrophenol + propionate | - |
? | |
alpha-naphthyl acetate + H2O | - |
Escherichia coli | alpha-naphthol + acetate | - |
? | |
alpha-naphthyl acetate + H2O | - |
Escherichia coli BW25113 | alpha-naphthol + acetate | - |
? | |
additional information | no activity by FrmB and YeiG with S-(1,2-dicarboxyethyl)glutathione, S-hexylglutathione, and S-(4-nitrobenzyl)glutathione | Escherichia coli | ? | - |
? | |
additional information | no activity by FrmB and YeiG with S-(1,2-dicarboxyethyl)glutathione, S-hexylglutathione, and S-(4-nitrobenzyl)glutathione | Escherichia coli BW25113 | ? | - |
? | |
S-formylglutathione + H2O | the substrate is an intermediate in the glutathione-dependent formaldehyde detoxification pathway | Escherichia coli | glutathione + formate | - |
? | |
S-formylglutathione + H2O | best substrate of FrmB and YeiG | Escherichia coli | glutathione + formate | - |
? | |
S-formylglutathione + H2O | the substrate is an intermediate in the glutathione-dependent formaldehyde detoxification pathway | Escherichia coli BW25113 | glutathione + formate | - |
? | |
S-lactoylglutathione + H2O | substrate of FrmB and YeiG | Escherichia coli | glutathione + lactate | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | structure analysis in comparison to the Saccharomyces cerevisiae crystal structure, PDB code 1pv1, the enzyme shows a canonical alpha/beta-hydrolase fold | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
FrmB | - |
Escherichia coli |
YeiG | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
S-Lactoylglutathione | pH 8.0, 37°C, recombinant FrmB | Escherichia coli | |
0.14 | - |
4-nitrophenyl propionate | pH 8.0, 37°C, recombinant FrmB | Escherichia coli | |
0.26 | - |
4-nitrophenyl acetate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
0.3 | - |
4-nitrophenyl acetate | pH 8.0, 37°C, recombinant FrmB | Escherichia coli | |
0.36 | - |
4-nitrophenyl butyrate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
0.37 | - |
4-nitrophenyl propionate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
0.49 | - |
4-nitrophenyl caproate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
1.02 | - |
S-Lactoylglutathione | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
2.17 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant D80A | Escherichia coli | |
2.23 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant D255A | Escherichia coli | |
2.96 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant D199A | Escherichia coli | |
3.13 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant D218A | Escherichia coli | |
3.3 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
3.81 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant D218A | Escherichia coli | |
5.09 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
5.1 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant C54A | Escherichia coli | |
5.12 | - |
Alpha-naphthyl acetate | pH 8.0, 37°C, recombinant YeiG mutant C26A | Escherichia coli | |
6.51 | - |
S-formylglutathione | pH 8.0, 37°C, recombinant wild-type YeiG | Escherichia coli | |
28.5 | - |
S-formylglutathione | pH 8.0, 37°C, recombinant FrmB | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |