Cloned (Comment) | Organism |
---|---|
gldABC encoding for wild-type and mutant GDHt amplified, PCR fragment digested with NcoI and HindIII, inserted into pSE380 linearized with the same enzymes and transforming the competent cells of Escherichia coli JM109 | Klebsiella pneumoniae |
Protein Variants | Comment | Organism |
---|---|---|
I498A | optimal pH and optimal temperature as the wild-type. KM for glycerol comparable to the wild-type | Klebsiella pneumoniae |
I498C | active, optimal pH as the wild-type, lower optimal temperature as the wild-type. KM for glycerol comparable to the wild-type | Klebsiella pneumoniae |
I498D | active, lower optimal pH as the wild-type, optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
I498M | active, lower optimal pH and lower optimal temperature as the wild-type. Lower KM for glycerol as the wild-type | Klebsiella pneumoniae |
I498N | active, optimal pH as the wild-type, lower optimal temperature as the wild-type. Lower KM for glycerol as the wild-type | Klebsiella pneumoniae |
I498P | active, lower optimal pH and lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
I498S | active, lower optimal pH and lower optimal temperature as the wild-type. KM for glycerol comparable to the wild-type | Klebsiella pneumoniae |
I498T | active, lower optimal pH as the wild-type, optimal temperature as the wild-type. Lower KM for glycerol as the wild-type | Klebsiella pneumoniae |
I498V | alpha subunit has half-life of 11 h in contrast to 1 h for the wild-type. Optimal pH and optimal temperature as the wild-type. KM for glycerol comparable to the wild-type | Klebsiella pneumoniae |
I498V/Q42L | alpha subunit of I498V and beta subunit of Q42L have half-life of 16 h in contrast to 1 h for the wild-type | Klebsiella pneumoniae |
I498W | active, lower optimal pH and lower optimal temperature as the wild-type. KM for glycerol comparable to the wild-type | Klebsiella pneumoniae |
Q42A | active, lower optimal pH and lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42C | active, lower optimal pH and lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42D | most affected, alpha-helix around 152 positions is totally distorted. Higher optimal pH as the wild-type, lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42E | active, lower optimal pH and lower optimal temperature as the wild-type. KM for glycerol comparable to the wild-type | Klebsiella pneumoniae |
Q42F | most active one, catalytic activity in beta-subunit is 8.3fold higher than the wild-type. Higher optimal pH and higher optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42G | active, lower optimal pH and lower optimal temperature as the wild-type. KM for glycerol comparable to the wild-type | Klebsiella pneumoniae |
Q42H | active, optimal pH as the wild-type, higher optimal temperature as the wild-type. KM for glycerol comparable to the wild-type | Klebsiella pneumoniae |
Q42I | active, lower optimal pH and lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42K | active, higher optimal pH and lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42L | lower optimal pH and lower optimal temperature as the wild-type. Enzyme efficiency in beta-subunit for substrate glycerol is 336fold higher than that for 1,2-propanediol. Mutation at position of the beta-subunit contributes to the increase of the thermostability as well as the pH stability of the enzyme. Lower KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42M | active, lower optimal pH and lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42N | most affected, alpha-helix around 152 positions is totally distorted. Higher optimal pH and lower optimal temperature as the wild-type. KM for glycerol comparable to the wild-type | Klebsiella pneumoniae |
Q42P | active, optimal pH as the wild-type, lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42R | most affected, alpha-helix around 152 positions is totally distorted. Higher optimal pH and lower optimal temperature as the wild-type. Lower KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42S | enzyme efficiency in beta-subunit for substrate glycerol is 80fold higher than that for 1,2-propanediol. Higher optimal pH and higher optimal temperature as the wild-type. Lower KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42T | active, lower optimal pH and lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42V | active, lower optimal pH and lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42W | active, lower optimal pH and lower optimal temperature as the wild-type. Lower KM for glycerol as the wild-type | Klebsiella pneumoniae |
Q42Y | active, lower optimal pH and lower optimal temperature as the wild-type. Higher KM for glycerol as the wild-type | Klebsiella pneumoniae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.09 | - |
1,2-propanediol | mutant I498N, pH 9.0, 42°C | Klebsiella pneumoniae | |
0.12 | - |
1,2-propanediol | mutant I498M, pH 8.0, 42°C | Klebsiella pneumoniae | |
0.14 | - |
1,2-propanediol | mutant Q42R, pH 9.5, 37°C | Klebsiella pneumoniae | |
0.17 | - |
1,2-propanediol | mutant I498V, pH 9.0, 45°C | Klebsiella pneumoniae | |
0.19 | - |
1,2-propanediol | mutant I498W, pH 8.0, 42°C. Mutant Q42D, pH 9.5, 42°C, mutant Q42E, pH 8.0, 42°C. Mutant Q42F, pH 9.5, 47°C. Mutant Q42N, pH 9.5, 42°C. Mutant Q42Y, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.24 | - |
1,2-propanediol | mutant I498C, pH 9.0, 40°C | Klebsiella pneumoniae | |
0.24 | - |
1,2-propanediol | mutant Q42M, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.24 | - |
1,2-propanediol | wild-type, pH 9.0, 45°C | Klebsiella pneumoniae | |
0.27 | - |
1,2-propanediol | mutant Q42K, pH 9.5, 42°C | Klebsiella pneumoniae | |
0.31 | - |
1,2-propanediol | mutant Q42P, pH 9.0, 42°C | Klebsiella pneumoniae | |
0.37 | - |
1,2-propanediol | mutant I498S, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.4 | - |
glycerol | mutant Q42L, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.44 | - |
glycerol | mutant Q42R, pH 9.5, 37°C | Klebsiella pneumoniae | |
0.45 | - |
1,2-propanediol | mutant Q42S, pH 9.5, 46.7°C | Klebsiella pneumoniae | |
0.48 | - |
glycerol | mutant Q42S, pH 9.5, 46.7°C | Klebsiella pneumoniae | |
0.5 | - |
glycerol | mutant I498M, pH 8.0, 42°C | Klebsiella pneumoniae | |
0.52 | - |
glycerol | mutant I498T, pH 8.5, 45°C | Klebsiella pneumoniae | |
0.55 | - |
glycerol | mutant I498N, pH 9.0, 42°C | Klebsiella pneumoniae | |
0.55 | - |
1,2-propanediol | mutant Q42H, pH 9.0, 47°C | Klebsiella pneumoniae | |
0.57 | - |
glycerol | mutant Q42W, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.58 | - |
glycerol | mutant I498A, pH 9.0, 45°C | Klebsiella pneumoniae | |
0.59 | - |
glycerol | mutant I498V, pH 9.0, 45°C | Klebsiella pneumoniae | |
0.6 | - |
glycerol | wild-type, pH 9.0, 45°C. Mutant I498C, pH 9.0, 40°C. Mutant Q42G, pH 7.5, 42°C. Mutant Q42N, pH 9.5, 42°C | Klebsiella pneumoniae | |
0.62 | - |
glycerol | mutant I498W, pH 8.0, 42°C | Klebsiella pneumoniae | |
0.62 | - |
glycerol | mutant Q42E, pH 8.0, 42°C | Klebsiella pneumoniae | |
0.64 | - |
1,2-propanediol | mutant Q42I, pH 8.5, 37°C | Klebsiella pneumoniae | |
0.65 | - |
1,2-propanediol | mutant Q42G, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.66 | - |
glycerol | mutant Q42H, pH 9.0, 47°C | Klebsiella pneumoniae | |
0.67 | - |
glycerol | mutant I498S, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.7 | - |
glycerol | mutant Q42P, pH 9.0, 42°C | Klebsiella pneumoniae | |
0.75 | - |
1,2-propanediol | mutant Q42A, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.77 | - |
1,2-propanediol | mutant Q42L, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.82 | - |
glycerol | mutant Q42T, pH 7.5, 42°C | Klebsiella pneumoniae | |
0.9 | - |
glycerol | mutant I498P, pH 8.0, 42°C | Klebsiella pneumoniae | |
0.95 | - |
1,2-propanediol | mutant I498A, pH 9.0, 45°C | Klebsiella pneumoniae | |
1 | - |
1,2-propanediol | mutant I498D, pH 8.0, 45°C | Klebsiella pneumoniae | |
1.03 | - |
glycerol | mutant Q42Y, pH 7.5, 42°C | Klebsiella pneumoniae | |
1.07 | - |
glycerol | mutant Q42I, pH 8.5, 37°C | Klebsiella pneumoniae | |
1.13 | - |
1,2-propanediol | mutant Q42C, pH 8.0, 42°C | Klebsiella pneumoniae | |
1.18 | - |
glycerol | mutant Q42M, pH 7.5, 42°C | Klebsiella pneumoniae | |
1.22 | - |
glycerol | mutant Q42C, pH 8.0, 42°C | Klebsiella pneumoniae | |
1.3 | - |
glycerol | mutant Q42V, pH 7.5, 42°C | Klebsiella pneumoniae | |
1.38 | - |
1,2-propanediol | mutant Q42T, pH 7.5, 42°C | Klebsiella pneumoniae | |
1.5 | - |
glycerol | mutant I498D, pH 8.0, 45°C | Klebsiella pneumoniae | |
1.54 | - |
1,2-propanediol | mutant Q42V, pH 7.5, 42°C | Klebsiella pneumoniae | |
1.8 | - |
glycerol | mutant Q42A, pH 7.5, 42°C | Klebsiella pneumoniae | |
1.9 | - |
1,2-propanediol | mutant I498P, pH 8.0, 42°C | Klebsiella pneumoniae | |
1.94 | - |
glycerol | mutant Q42D, pH 9.5, 42°C | Klebsiella pneumoniae | |
2.61 | - |
glycerol | mutant Q42F, pH 9.5, 47°C | Klebsiella pneumoniae | |
2.8 | - |
glycerol | mutant Q42K, pH 9.5, 42°C | Klebsiella pneumoniae | |
3.3 | - |
1,2-propanediol | mutant I498T, pH 8.5, 45°C | Klebsiella pneumoniae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Klebsiella pneumoniae | - |
- |
- |
Klebsiella pneumoniae 10018 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
by Ni-nitrilotriacetate affinity chromatography followed by gel filtration | Klebsiella pneumoniae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,2-propanediol | - |
Klebsiella pneumoniae | propionaldehyde + H2O | - |
? | |
1,2-propanediol | - |
Klebsiella pneumoniae 10018 | propionaldehyde + H2O | - |
? | |
glycerol | reaction requires two steps, is catalyzed by GDHt and 1,3-PD oxidoreductase. GDHt is the rate-limiting enzyme in this process | Klebsiella pneumoniae | 3-hydroxypropanal + H2O | - |
? | |
glycerol | reaction requires two steps, is catalyzed by GDHt and 1,3-PD oxidoreductase. GDHt is the rate-limiting enzyme in this process | Klebsiella pneumoniae 10018 | 3-hydroxypropanal + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GDHt | - |
Klebsiella pneumoniae |
glycerol dehydratase | - |
Klebsiella pneumoniae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
mutant enzymes Q42I and Q42R | Klebsiella pneumoniae |
40 | - |
mutant enzyme I498C | Klebsiella pneumoniae |
42 | - |
mutant enzymes I498M, I498N, I498P, I498S, I498W, Q42A, Q42C, Q42D, Q42E, Q42G, Q42K, Q42L, Q42M, Q42N, Q42P, Q42T, Q42V, Q42W and Q42Y | Klebsiella pneumoniae |
45 | - |
wild-type enzyme and mutant enzymes I498A, I498D and I498T | Klebsiella pneumoniae |
46.7 | - |
mutant enzyme Q42S | Klebsiella pneumoniae |
47 | - |
mutant enzymes Q42F and Q42H | Klebsiella pneumoniae |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
t1/2: 1 h | Klebsiella pneumoniae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
mutant enzymes I498S, Q42A, Q42L, Q42M, Q42T, Q42V, Q42W, Q42Y and Q42G | Klebsiella pneumoniae |
8 | - |
mutant enzymes I498D, I498M, I498P, I498W, Q42C and Q42E | Klebsiella pneumoniae |
8.5 | - |
mutant enzymes I498T, Q42I and Q42E | Klebsiella pneumoniae |
9 | - |
wild-type enzyme and mutant enzymes I498C, I498A, I498N, I498V, Q42H and Q42P | Klebsiella pneumoniae |
9.5 | - |
mutant enzymes Q42N, Q42D, Q42F, Q42K, Q42R, Q42S, Q42Y and Q42G | Klebsiella pneumoniae |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6 | - |
t1/2: 1 h | Klebsiella pneumoniae |
8 | - |
t1/2: 1 h | Klebsiella pneumoniae |