Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 AI or NEB 10beta cells | Basidiomycota sp. |
Crystallization (Comment) | Organism |
---|---|
sitting drop vapor diffusion method, using 1.6-1.95 M ammonium sulfate and 0.1 M MES or 0.1 M citrate buffer pH 6.5 | Basidiomycota sp. |
Protein Variants | Comment | Organism |
---|---|---|
D222A | the mutant shows reduced catalytic efficiency with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), Reactive Black 5 and H2O, as compared to the wild type enzyme | Basidiomycota sp. |
D222E | the mutant shows increased catalytic efficiency with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and reduced activity with Reactive Black 5 and H2O, as compared to the wild type enzyme | Basidiomycota sp. |
D222E/R390K | the mutant shows increased catalytic efficiency with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and reduced activity with Reactive Black 5 and H2O, as compared to the wild type enzyme | Basidiomycota sp. |
R390A | the mutant shows reduced catalytic efficiency with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), Reactive Black 5 and H2O, as compared to the wild type enzyme | Basidiomycota sp. |
R390K | the mutant shows increased catalytic efficiency with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and H2O and reduced activity with Reactive Black 5, as compared to the wild type enzyme | Basidiomycota sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | - |
Basidiomycota sp. | |
Direct blue 71 | - |
Basidiomycota sp. | |
Reactive Blue 19 | - |
Basidiomycota sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00023 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.0029 | - |
Direct Red 80 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.0033 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.0042 | - |
Reactive Black 5 | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.006 | - |
Reactive Black 5 | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.0068 | - |
Reactive Red 2 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.0089 | - |
Reactive Black 5 | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.01 | - |
Reactive Black 5 | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.011 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.013 | - |
Reactive Black 5 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.013 | - |
Direct blue 71 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.013 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.013 | - |
Reactive Black 5 | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.015 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.018 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.031 | - |
H2O2 | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.039 | - |
H2O2 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.042 | - |
Reactive Blue 19 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.061 | - |
H2O2 | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.095 | - |
H2O2 | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. | |
1.93 | - |
H2O2 | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
4.18 | - |
H2O2 | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. | |
80 | - |
Mn2+ | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Basidiomycota sp. | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-Sepharose column chromatography and Superdex 75 gel filtration | Basidiomycota sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + H2O2 + H+ | - |
Basidiomycota sp. | oxidized 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + H2O | - |
? | |
Direct Blue 71 + H2O2 + H+ | - |
Basidiomycota sp. | oxidized Direct Blue 71 + H2O | - |
? | |
Direct Red 80 + H2O2 + H+ | - |
Basidiomycota sp. | oxidized Direct Red 80 + H2O | - |
? | |
Mn2+ + H2O2 + H+ | - |
Basidiomycota sp. | Mn3+ + 2 H2O | - |
? | |
additional information | no activity with thioanisole | Basidiomycota sp. | ? | - |
- |
|
Reactive Black 5 + H2O2 + H+ | - |
Basidiomycota sp. | oxidized Reactive Black 5 + H2O | - |
? | |
Reactive Blue 19 + H2O2 + H+ | - |
Basidiomycota sp. | oxidized Reactive Blue 19 + H2O | - |
? | |
Reactive Red 2 + H2O2 + H+ | - |
Basidiomycota sp. | oxidized Reactive Red 2 + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 68992, SUMO-fused enzyme, SDS-PAGE | Basidiomycota sp. |
? | x * 69000, calculated from amino acid sequence | Basidiomycota sp. |
Synonyms | Comment | Organism |
---|---|---|
AncDyPD-b1 | - |
Basidiomycota sp. |
DyP | - |
Basidiomycota sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
60% activity loss is observed after 1 h incubation at 40°C | Basidiomycota sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.002 | - |
Reactive Black 5 | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.015 | - |
Reactive Black 5 | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.036 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.038 | - |
Reactive Black 5 | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.04 | - |
Direct Red 80 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.11 | - |
Reactive Black 5 | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.12 | - |
Reactive Black 5 | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.14 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.19 | - |
Reactive Red 2 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.2 | - |
Direct blue 71 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.28 | - |
H2O2 | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.47 | - |
H2O2 | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.5 | - |
Mn2+ | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.58 | - |
Reactive Black 5 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
1.6 | - |
H2O2 | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
2.5 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. | |
4.3 | - |
H2O2 | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. | |
5 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
5.4 | - |
H2O2 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
5.4 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
7.1 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
13.6 | - |
H2O2 | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
22 | - |
Reactive Blue 19 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3 | 4 | there is no significant activity at pH 7.0 | Basidiomycota sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
heme | - |
Basidiomycota sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
Direct blue 71 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.032 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.047 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.072 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.078 | - |
Reactive Blue 19 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.079 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.09 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.19 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0063 | - |
Mn2+ | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.11 | - |
H2O2 | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.14 | - |
H2O2 | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
0.45 | - |
Reactive Black 5 | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
1.5 | - |
Reactive Black 5 | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
4.2 | - |
Reactive Black 5 | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. | |
8.9 | - |
Reactive Black 5 | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
12 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. | |
13 | - |
Direct Red 80 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
15 | - |
Direct blue 71 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
18 | - |
Reactive Black 5 | mutant enzyme R390A, at pH 3.0 and 25°C | Basidiomycota sp. | |
26 | - |
H2O2 | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
28 | - |
Reactive Red 2 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
45 | - |
H2O2 | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. | |
46 | - |
Reactive Black 5 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
140 | - |
H2O2 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
150 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222A, at pH 3.0 and 25°C | Basidiomycota sp. | |
290 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
330 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222E/R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
440 | - |
H2O2 | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
530 | - |
Reactive Blue 19 | wild type enzyme, at pH 3.0 and 25°C | Basidiomycota sp. | |
530 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme R390K, at pH 3.0 and 25°C | Basidiomycota sp. | |
770 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | mutant enzyme D222E, at pH 3.0 and 25°C | Basidiomycota sp. |