BRENDA - Enzyme Database

Structural basis of the divergent oxygenation reactions catalyzed by the rieske nonheme iron oxygenase carbazole 1,9a-dioxygenase

Inoue, K.; Usami, Y.; Ashikawa, Y.; Noguchi, H.; Umeda, T.; Yamagami-Ashikawa, A.; Horisaki, T.; Uchimura, H.; Terada, T.; Nakamura, S.; Shimizu, K.; Habe, H.; Yamane, H.; Fujimoto, Z.; Nojiri, H.; Appl. Environ. Microbiol. 80, 2821-2832 (2014)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.14.12.22
expressed in Escherichia coli BL21(DE3) cells
Janthinobacterium sp. J3
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.14.12.22
hanging drop vapor diffusion method, using 0.1M ammonium acetate, 12.5% (w/v) polyethylene glycol 3350 in 0.05 M MES at pH 5.5-5.7
Janthinobacterium sp. J3
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.14.12.22
F275W
the mutant catalyzes the lateral dioxygenation of carbazole with higher yields than the wild type enzyme
Janthinobacterium sp. J3
1.14.12.22
I262V
the mutant catalyzes the lateral dioxygenation of carbazole with higher yields than the wild type enzyme
Janthinobacterium sp. J3
1.14.12.22
Q282Y
the mutant catalyzes the lateral dioxygenation of carbazole with higher yields than the wild type enzyme
Janthinobacterium sp. J3
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.12.22
0.0507
-
9H-carbazole
mutant enzyme I262V, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
0.0546
-
9H-carbazole
mutant enzyme Q282N, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
0.0548
-
9H-carbazole
mutant enzyme Q282Y, at pH 7.5 and 30°C
Janthinobacterium sp. J3
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.12.22
9H-carbazole + NADH + H+ + O2
Janthinobacterium sp. J3
-
2'-aminobiphenyl-2,3-diol + NAD+
-
-
?
1.14.12.22
9H-carbazole + NADPH + H+ + O2
Janthinobacterium sp. J3
-
2'-aminobiphenyl-2,3-diol + NADP+
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.12.22
Janthinobacterium sp. J3
Q84II6
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.14.12.22
HiTrap chelation column chromatography and Superdex 200 gel filtration
Janthinobacterium sp. J3
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.12.22
9H-carbazole + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
2'-aminobiphenyl-2,3-diol + NAD+
-
-
-
?
1.14.12.22
9H-carbazole + NADPH + H+ + O2
-
741682
Janthinobacterium sp. J3
2'-aminobiphenyl-2,3-diol + NADP+
-
-
-
?
1.14.12.22
anthracene + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
biphenyl + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
dibenzo-4-dioxin + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
dibenzothiophene + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
fluorene + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
naphthalene + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.14.12.22
4
-
9H-carbazole
mutant enzyme Q282N, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
4.62
-
9H-carbazole
mutant enzyme I262V, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
5.49
-
9H-carbazole
mutant enzyme Q282Y, at pH 7.5 and 30°C
Janthinobacterium sp. J3
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.14.12.22
NADH
-
Janthinobacterium sp. J3
1.14.12.22
NADPH
-
Janthinobacterium sp. J3
1.14.12.22
[2Fe-2S]-center
-
Janthinobacterium sp. J3
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.12.22
expressed in Escherichia coli BL21(DE3) cells
Janthinobacterium sp. J3
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.14.12.22
NADH
-
Janthinobacterium sp. J3
1.14.12.22
NADPH
-
Janthinobacterium sp. J3
1.14.12.22
[2Fe-2S]-center
-
Janthinobacterium sp. J3
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.14.12.22
hanging drop vapor diffusion method, using 0.1M ammonium acetate, 12.5% (w/v) polyethylene glycol 3350 in 0.05 M MES at pH 5.5-5.7
Janthinobacterium sp. J3
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.14.12.22
F275W
the mutant catalyzes the lateral dioxygenation of carbazole with higher yields than the wild type enzyme
Janthinobacterium sp. J3
1.14.12.22
I262V
the mutant catalyzes the lateral dioxygenation of carbazole with higher yields than the wild type enzyme
Janthinobacterium sp. J3
1.14.12.22
Q282Y
the mutant catalyzes the lateral dioxygenation of carbazole with higher yields than the wild type enzyme
Janthinobacterium sp. J3
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.12.22
0.0507
-
9H-carbazole
mutant enzyme I262V, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
0.0546
-
9H-carbazole
mutant enzyme Q282N, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
0.0548
-
9H-carbazole
mutant enzyme Q282Y, at pH 7.5 and 30°C
Janthinobacterium sp. J3
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.12.22
9H-carbazole + NADH + H+ + O2
Janthinobacterium sp. J3
-
2'-aminobiphenyl-2,3-diol + NAD+
-
-
?
1.14.12.22
9H-carbazole + NADPH + H+ + O2
Janthinobacterium sp. J3
-
2'-aminobiphenyl-2,3-diol + NADP+
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.12.22
HiTrap chelation column chromatography and Superdex 200 gel filtration
Janthinobacterium sp. J3
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.12.22
9H-carbazole + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
2'-aminobiphenyl-2,3-diol + NAD+
-
-
-
?
1.14.12.22
9H-carbazole + NADPH + H+ + O2
-
741682
Janthinobacterium sp. J3
2'-aminobiphenyl-2,3-diol + NADP+
-
-
-
?
1.14.12.22
anthracene + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
biphenyl + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
dibenzo-4-dioxin + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
dibenzothiophene + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
fluorene + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
1.14.12.22
naphthalene + NADH + H+ + O2
-
741682
Janthinobacterium sp. J3
?
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.14.12.22
4
-
9H-carbazole
mutant enzyme Q282N, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
4.62
-
9H-carbazole
mutant enzyme I262V, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
5.49
-
9H-carbazole
mutant enzyme Q282Y, at pH 7.5 and 30°C
Janthinobacterium sp. J3
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.14.12.22
0.000733
-
9H-carbazole
mutant enzyme Q282N, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
0.00091
-
9H-carbazole
mutant enzyme I262V, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
0.001
-
9H-carbazole
mutant enzyme Q282Y, at pH 7.5 and 30°C
Janthinobacterium sp. J3
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.14.12.22
0.000733
-
9H-carbazole
mutant enzyme Q282N, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
0.00091
-
9H-carbazole
mutant enzyme I262V, at pH 7.5 and 30°C
Janthinobacterium sp. J3
1.14.12.22
0.001
-
9H-carbazole
mutant enzyme Q282Y, at pH 7.5 and 30°C
Janthinobacterium sp. J3