Application | Comment | Organism |
---|---|---|
biofuel production | attractive oxidoreductase for bioelectrochemical applications. Its two-domain structure allows the flavoheme enzyme to establish direct electron transfer to biosensor and biofuel cell electrodes. The application of CDH in these devices is impeded by its limited stability under turnover conditions. Screening results in one CDH variant that exhibits improved turnover stability on a biosensor electrode, which is suitable for the application in implantable continuous glucose monitoring biosensors or biofuel cells | Thermothelomyces myriococcoides |
Protein Variants | Comment | Organism |
---|---|---|
L324C | the variant loses approximately half of their activities within 52 h at 37°C | Thermothelomyces myriococcoides |
L324C/M409S | lower catalytic activity as compared to wild-type enzyme. The variant loses approximately half of their activities within 52 h at 37°C | Thermothelomyces myriococcoides |
M309K | retains most of its initial activity among the single variants. Mutation improve the turnover stability of the enzyme | Thermothelomyces myriococcoides |
M309K/L324C | the Tm-value of the variant is 1.8°C lower than the wild-type value | Thermothelomyces myriococcoides |
M309K/M409S | variant with the highest residual activity after 4 h at 41 °C under turnover conditions. Lower catalytic activity as compared to wild-type enzyme | Thermothelomyces myriococcoides |
M309V | the mutant enzyme shows significant stability improvements compared to the wild-type enzyme. Higher cytochrome c/2,6-dichloroindophenol activity ratio than the wild-type enzyme | Thermothelomyces myriococcoides |
M409S | mutation improve the turnover stability of the enzyme. Most stable variant with an electrode half-life of 13.5 h | Thermothelomyces myriococcoides |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | 3 | cytochrome c | pH 7.4, 37°C, mutant enzyme M309K/L324C | Thermothelomyces myriococcoides | |
9 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme M309K/M409S | Thermothelomyces myriococcoides | |
11 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme L324C/M409S | Thermothelomyces myriococcoides | |
41 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme M409S | Thermothelomyces myriococcoides | |
44 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M309K/M409S | Thermothelomyces myriococcoides | |
72 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme M309K | Thermothelomyces myriococcoides | |
81 | - |
cytochrome c | pH 7.4, 37°C, wild-type enzyme | Thermothelomyces myriococcoides | |
84 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M409S | Thermothelomyces myriococcoides | |
118 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme L324C | Thermothelomyces myriococcoides | |
130 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme M309V | Thermothelomyces myriococcoides | |
138 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, wild-type enzyme | Thermothelomyces myriococcoides | |
196 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M309V | Thermothelomyces myriococcoides | |
200 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M309K | Thermothelomyces myriococcoides | |
205 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme L324C | Thermothelomyces myriococcoides | |
219 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme L324C/M409S | Thermothelomyces myriococcoides | |
350 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M309K/L324C | Thermothelomyces myriococcoides |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermothelomyces myriococcoides | A9XK88 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glucose + 2,6-dichloroindophenol | - |
Thermothelomyces myriococcoides | ? + reduced 2,6-dichloroindophenol | - |
? | |
glucose + cytochrome c | - |
Thermothelomyces myriococcoides | ? + reduced cytochrome c | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Cdh | - |
Thermothelomyces myriococcoides |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
residual activity after 52 h when stored in 11 mM PBS pH 7.4 buffer: wild-type enzyme (106%), mutant enzyme M309V: (103%), mutant enzyme M309K: (94%), mutant enzyme M409S: (102%), mutant enzyme M309K/M409S: (116%), mutant enzyme L324C: (57%), mutant enzyme L324C/M409S: (58%), mutant enzyme M309K/L324C: (8%) | Thermothelomyces myriococcoides |
47.6 | - |
Tm-value, mutant enzyme M309K/M409S | Thermothelomyces myriococcoides |
49.8 | - |
Tm-value, mutant enzyme M409S | Thermothelomyces myriococcoides |
51.9 | - |
Tm-value, mutant enzyme L324C/M409S | Thermothelomyces myriococcoides |
53.9 | - |
Tm-value, mutant enzyme M309K | Thermothelomyces myriococcoides |
55 | - |
Tm-value, mutant enzyme M309K/L324C | Thermothelomyces myriococcoides |
56.3 | - |
Tm-value, mutant enzyme L324C | Thermothelomyces myriococcoides |
56.4 | - |
Tm-value, mutant enzyme M309V | Thermothelomyces myriococcoides |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.5 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme M309K/M409S | Thermothelomyces myriococcoides | |
1.2 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme L324C/M409S | Thermothelomyces myriococcoides | |
1.2 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme M309K/L324C | Thermothelomyces myriococcoides | |
1.6 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M309K/M409S | Thermothelomyces myriococcoides | |
3 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme M409S | Thermothelomyces myriococcoides | |
3.1 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme L324C | Thermothelomyces myriococcoides | |
3.1 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M309K/L324C | Thermothelomyces myriococcoides | |
3.7 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme L324C/M409S | Thermothelomyces myriococcoides | |
4.2 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme M309V | Thermothelomyces myriococcoides | |
6.2 | - |
cytochrome c | pH 7.4, 37°C, mutant enzyme M309K | Thermothelomyces myriococcoides | |
6.3 | - |
cytochrome c | pH 7.4, 37°C, wild-type enzyme | Thermothelomyces myriococcoides | |
7.1 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M409S | Thermothelomyces myriococcoides | |
8.6 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme L324C | Thermothelomyces myriococcoides | |
13 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M309K | Thermothelomyces myriococcoides | |
15.1 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, mutant enzyme M309V | Thermothelomyces myriococcoides | |
17.4 | - |
2,6-dichloroindophenol | pH 7.4, 37°C, wild-type enzyme | Thermothelomyces myriococcoides |