Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BLR(DE3) cells | Pseudomonas fluorescens |
Protein Variants | Comment | Organism |
---|---|---|
D230A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
E133A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
E133Q | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
H303A | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
H303A/R373A/K381A | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
K295A | inactive | Pseudomonas fluorescens |
K381A | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
N191A | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
N300A | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
N300D | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
N300S | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
R373A | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas fluorescens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.56 | - |
D-mannitol | mutant enzyme N191A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
5.98 | - |
D-mannitol | wild type enzyme, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
8.15 | - |
D-mannitol | mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
14.96 | - |
D-mannitol | mutant enzyme E133Q, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
17.49 | - |
D-mannitol | mutant enzyme R373A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
18.21 | - |
D-mannitol | mutant enzyme E133A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
20.36 | - |
D-arabitol | mutant enzyme D230A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
20.42 | - |
D-mannitol | mutant enzyme N300D, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
23.89 | - |
D-mannitol | mutant enzyme K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
33.15 | - |
D-arabitol | mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
42.74 | - |
D-mannitol | mutant enzyme D230A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
49.95 | - |
D-mannitol | mutant enzyme N300S, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
55.85 | - |
D-arabitol | wild type enzyme, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
68.94 | - |
D-mannitol | mutant enzyme N300A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
93.85 | - |
D-mannitol | mutant enzyme H303A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
94.82 | - |
D-arabitol | mutant enzyme N300D, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
125.5 | - |
D-arabitol | mutant enzyme N191A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
126.5 | - |
D-arabitol | mutant enzyme N300S, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
126.9 | - |
D-arabitol | mutant enzyme E133Q, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
132.1 | - |
D-arabitol | mutant enzyme N300A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
134.1 | - |
D-arabitol | mutant enzyme E133A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
144.5 | - |
D-arabitol | mutant enzyme R373A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
145.1 | - |
D-arabitol | mutant enzyme H303A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
176.9 | - |
D-arabitol | mutant enzyme K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
1800 | - |
meso-erythritol | Km far above 1800 mM, mutant enzyme E133A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
1800 | - |
meso-erythritol | Km far above 1800 mM, mutant enzyme E133Q, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
1800 | - |
meso-erythritol | Km far above 1800 mM, mutant enzyme H303A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
1800 | - |
meso-erythritol | Km far above 1800 mM, mutant enzyme N191A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
1800 | - |
meso-erythritol | Km far above 1800 mM, mutant enzyme N300A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
1800 | - |
meso-erythritol | Km far above 1800 mM, mutant enzyme N300S, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
1800 | - |
meso-erythritol | Km far above 1800 mM, wild type enzyme, at pH 7.4 and 25°C | Pseudomonas fluorescens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-mannitol + NAD+ | Pseudomonas fluorescens | - |
D-fructose + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas fluorescens | O08355 | - |
- |
Purification (Comment) | Organism |
---|---|
HisPur cobalt resin column chromatography | Pseudomonas fluorescens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-arabitol + NAD+ | - |
Pseudomonas fluorescens | ? | - |
? | |
D-mannitol + NAD+ | - |
Pseudomonas fluorescens | D-fructose + NADH + H+ | - |
r | |
meso-erythritol + NAD+ | - |
Pseudomonas fluorescens | ? | - |
? | |
additional information | no activity with glycerol, 1, 2-hexanediol, and 1,2,3-hexanetriol | Pseudomonas fluorescens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MDH | - |
Pseudomonas fluorescens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00083 | - |
D-mannitol | mutant enzyme N300D, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.001 | - |
D-mannitol | mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0012 | - |
meso-erythritol | mutant enzyme N191A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0013 | - |
D-arabitol | mutant enzyme N300D, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0017 | - |
D-arabitol | mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0024 | - |
meso-erythritol | mutant enzyme N300S, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0029 | - |
D-arabitol | mutant enzyme D230A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0036 | - |
meso-erythritol | mutant enzyme N300A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0159 | - |
meso-erythritol | mutant enzyme H303A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.018 | - |
D-mannitol | mutant enzyme R373A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.034 | - |
D-arabitol | mutant enzyme R373A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.044 | - |
D-arabitol | mutant enzyme N191A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.06 | - |
D-arabitol | mutant enzyme N300S, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.092 | - |
D-arabitol | mutant enzyme N300A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.096 | - |
D-mannitol | mutant enzyme N300S, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.19 | - |
D-mannitol | mutant enzyme K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.209 | - |
D-mannitol | mutant enzyme N191A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.21 | - |
D-mannitol | mutant enzyme N300A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.29 | - |
D-arabitol | mutant enzyme H303A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.37 | - |
D-arabitol | mutant enzyme K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.39 | - |
D-mannitol | mutant enzyme D230A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.489 | - |
meso-erythritol | mutant enzyme E133Q, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.49 | - |
D-mannitol | mutant enzyme H303A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.538 | - |
meso-erythritol | mutant enzyme E133A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.599 | - |
meso-erythritol | wild type enzyme, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
3.22 | - |
D-mannitol | mutant enzyme E133Q, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
3.99 | - |
D-mannitol | mutant enzyme E133A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
4.39 | - |
D-mannitol | wild type enzyme, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
5.5 | - |
D-arabitol | mutant enzyme E133Q, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
6.59 | - |
D-arabitol | wild type enzyme, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
7.3 | - |
D-arabitol | mutant enzyme E133A, at pH 7.4 and 25°C | Pseudomonas fluorescens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Pseudomonas fluorescens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0000021 | - |
meso-erythritol | mutant enzyme N191A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0000044 | - |
meso-erythritol | mutant enzyme N300S, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0000065 | - |
meso-erythritol | mutant enzyme N300A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.000014 | - |
D-arabitol | mutant enzyme N300D, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.000029 | - |
meso-erythritol | mutant enzyme H303A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00004 | - |
D-mannitol | mutant enzyme N300D, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.000051 | - |
D-arabitol | mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00014 | - |
D-arabitol | mutant enzyme D230A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00016 | - |
D-mannitol | mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00024 | - |
D-arabitol | mutant enzyme R373A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00035 | - |
D-arabitol | mutant enzyme N191A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00047 | - |
D-arabitol | mutant enzyme N300S, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00069 | - |
D-arabitol | mutant enzyme N300A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00088 | - |
meso-erythritol | mutant enzyme E133Q, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00097 | - |
meso-erythritol | mutant enzyme E133A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00102 | - |
D-mannitol | mutant enzyme R373A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.0018 | - |
meso-erythritol | wild type enzyme, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00192 | - |
D-mannitol | mutant enzyme N300S, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.002 | - |
D-arabitol | mutant enzyme H303A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00206 | - |
D-arabitol | mutant enzyme K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00311 | - |
D-mannitol | mutant enzyme N300A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00458 | - |
D-mannitol | mutant enzyme N191A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00522 | - |
D-mannitol | mutant enzyme H303A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00775 | - |
D-mannitol | mutant enzyme K381A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.00908 | - |
D-mannitol | mutant enzyme D230A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.04332 | - |
D-arabitol | mutant enzyme E133Q, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.05448 | - |
D-arabitol | mutant enzyme E133A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.118 | - |
D-arabitol | wild type enzyme, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.215 | - |
D-mannitol | mutant enzyme E133Q, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.219 | - |
D-mannitol | mutant enzyme E133A, at pH 7.4 and 25°C | Pseudomonas fluorescens | |
0.734 | - |
D-mannitol | wild type enzyme, at pH 7.4 and 25°C | Pseudomonas fluorescens |