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Literature summary for 4.2.1.30 extracted from

  • Qi, X.; Chen, Y.; Jiang, K.; Zuo, W.; Luo, Z.; Wei, Y.; Du, L.; Wei, H.; Huang, R.; Du, Q.
    Saturation-mutagenesis in two positions distant from active site of a Klebsiella pneumoniae glycerol dehydratase identifies some highly active mutants (2009), J. Biotechnol., 144, 43-50.
    View publication on PubMed
Search for more literature for 4.2.1.30

Cloned(Commentary)

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

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 [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

Organism UniProt Comment Textmining
Klebsiella pneumoniae
-
-
-
Klebsiella pneumoniae 10018
-
-
-

Purification (Commentary)

Purification (Comment) Organism
by Ni-nitrilotriacetate affinity chromatography followed by gel filtration Klebsiella pneumoniae

Substrates and Products (Substrate)

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

Synonyms Comment Organism
GDHt
-
 Klebsiella pneumoniae
glycerol dehydratase
-
 Klebsiella pneumoniae

Temperature Optimum [°C]

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 [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 t1/2: 1 h Klebsiella pneumoniae

pH Optimum

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 pH Stability Maximum Comment Organism
6
-
 t1/2: 1 h Klebsiella pneumoniae
8
-
 t1/2: 1 h Klebsiella pneumoniae