Cloned (Comment) | Organism |
---|---|
mutant genes F190Y, F190L, F190I and F190A are subcloned and expressed in Escherichia coli XL-1 Blue and DH5alphaF | Phanerodontia chrysosporium |
Crystallization (Comment) | Organism |
---|---|
crystal structure | Phanerodontia chrysosporium |
Protein Variants | Comment | Organism |
---|---|---|
F190A | mutant MnP: apparent Km-value for ferrocyanide oxidation is 1/8 of that for wild-type MnP and kcat is 4fold greater than that for wild-type enzyme, mutant enzyme is significantly destabilized to thermal denaturation, unstable at 37°C, rates of spontaneous reduction of the oxidized intermediates, compound I and II, are dramatically increased compared with those for the wild-type MnP | Phanerodontia chrysosporium |
F190I | mutant enzyme is significantly destabilized to thermal denaturation, unstable at 37°C, rates of spontaneous reduction of the oxidized intermediates, compound I and II, are 2fold greater than those for the wild-type MnP | Phanerodontia chrysosporium |
F190L | rates of spontaneous reduction of the oxidized intermediates, compound I and II, are 2fold greater than those for the wild-type MnP | Phanerodontia chrysosporium |
F190Y | engineered mutant | Phanerodontia chrysosporium |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.039 | - |
H2O2 | wild-type MnP1 | Phanerodontia chrysosporium | |
0.039 | 0.041 | H2O2 | MnP1 from mutants F190Y, F190L, F190I and F190A | Phanerodontia chrysosporium | |
0.074 | 0.08 | Mn2+ | MnP1 from mutants F190Y, F190L, F190I and F190A | Phanerodontia chrysosporium | |
0.083 | - |
Mn2+ | wild-type MnP1 | Phanerodontia chrysosporium | |
0.42 | - |
ferrocyanide | MnP1 from mutant F190A | Phanerodontia chrysosporium | |
0.42 | - |
ferrocyanide | mutant enzyme F190A, at pH 4.5 and 25°C | Phanerodontia chrysosporium | |
3.4 | 3.8 | ferrocyanide | MnP1 from mutants F190Y, F190L, F190I | Phanerodontia chrysosporium | |
3.4 | 3.8 | ferrocyanide | mutant enzyme F190I, at pH 4.5 and 25°C | Phanerodontia chrysosporium | |
3.5 | - |
ferrocyanide | wild-type MnP1 | Phanerodontia chrysosporium | |
3.5 | - |
ferrocyanide | mutant enzyme F190L, at pH 4.5 and 25 °C | Phanerodontia chrysosporium |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Phanerodontia chrysosporium | - |
- |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
46000 | - |
- |
Phanerodontia chrysosporium |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Phanerodontia chrysosporium | - |
- |
- |
Phanerodontia chrysosporium | - |
MnP1 | - |
Phanerodontia chrysosporium | - |
auxotrophic strain OGC107-1 | - |
Phanerodontia chrysosporium OGC101 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
purification of MnPs from the mutants F190Y, F190L, F190I and F190A | Phanerodontia chrysosporium |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ferrocyanide + H+ + H2O2 | - |
Phanerodontia chrysosporium | ferricyanide + H2O | - |
r | |
ferrocyanide + H+ + H2O2 | - |
Phanerodontia chrysosporium OGC101 | ferricyanide + H2O | - |
r | |
Mn2+ + H+ + H2O2 | unique binding and oxidation site for Mn2+, single Mn atom is hexacoordinate, with two water ligands and four carboxylate ligands from heme propionate 6 and amino acids Glu-35, Glu-39 and Asp-179 | Phanerodontia chrysosporium | Mn3+ + H2O | Mn3+ oxidizes lignin | ? | |
Mn2+ + H+ + H2O2 | unique binding and oxidation site for Mn2+, single Mn atom is hexacoordinate, with two water ligands and four carboxylate ligands from heme propionate 6 and amino acids Glu-35, Glu-39 and Asp-179 | Phanerodontia chrysosporium OGC101 | Mn3+ + H2O | Mn3+ oxidizes lignin | ? | |
additional information | catalytic cycle with oxidized intermediates MnP compound I and II | Phanerodontia chrysosporium | ? | - |
? | |
additional information | enzyme oxidizes ferrocyanide | Phanerodontia chrysosporium | ? | - |
? | |
additional information | catalytic cycle with oxidized intermediates MnP compound I and II | Phanerodontia chrysosporium OGC101 | ? | - |
? | |
additional information | enzyme oxidizes ferrocyanide | Phanerodontia chrysosporium OGC101 | ? | - |
? |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
49 | - |
wild-type MnP, MnP F190Y and MnP F190L: 330 s, 50% loss of activity, MnP F190I: 30 s, 50% loss of activity, MnP F190A: 5 s, complete loss of activity | Phanerodontia chrysosporium |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.4 | 4.2 | ferrocyanide | MnP1 from mutants F190Y, F190L, F190I | Phanerodontia chrysosporium | |
4 | - |
ferrocyanide | wild-type MnP1 | Phanerodontia chrysosporium | |
14.6 | - |
ferrocyanide | MnP1 from mutant F190A | Phanerodontia chrysosporium | |
240 | 290 | Mn2+ | MnP1 from mutants F190Y, F190L, F190I and F190A | Phanerodontia chrysosporium | |
290 | - |
Mn2+ | wild-type MnP1 | Phanerodontia chrysosporium |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
heme | heme environment, Phe-190 plays a critical role in stabilizing the heme environment, it acts as a steric barrier that protects the heme from reducing agents, increasing pH from 5.0 to 8.5 induces a Fe3+ high- to a low-spin transition | Phanerodontia chrysosporium |