BRENDA - Enzyme Database
show all sequences of 1.14.99.40

Active site residues critical for flavin binding and 5,6-dimethylbenzimidazole biosynthesis in the flavin destructase enzyme BluB

Yu, T.Y.; Mok, K.C.; Kennedy, K.J.; Valton, J.; Anderson, K.S.; Walker, G.C.; Taga, M.E.; Protein Sci. 21, 839-849 (2012)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
gene bluB, genotyping
Sinorhizobium meliloti
Engineering
Protein Variants
Commentary
Organism
A156V
site-directed mutagenesis, inactive mutant
Sinorhizobium meliloti
D32N
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Sinorhizobium meliloti
D32N/S167G
site-directed mutagenesis, the mutant shows no activity
Sinorhizobium meliloti
E78K
site-directed mutagenesis, the mutant is not soluble
Sinorhizobium meliloti
G110S
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
G133S
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
G193D
site-directed mutagenesis, inactive mutant
Sinorhizobium meliloti
G61D
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
M140I
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Sinorhizobium meliloti
M94I
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Sinorhizobium meliloti
P202L
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
P65L
site-directed mutagenesis, inactive mutant
Sinorhizobium meliloti
R30C
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
R30H
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
S167G
site-directed mutagenesis, the mutant shows higly reduced activity compared to the wild-type enzyme
Sinorhizobium meliloti
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
FMN/FMNH2 dissociation constants and free energy of binding of wild-type and mutant enzymes, overview
Sinorhizobium meliloti
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
FMNH2 + NADH + H+ + O2
Sinorhizobium meliloti
-
5,6-dimethylbenzimidazole + D-erythrose 4-phosphate + NAD+ + ?
5,6-dimethylbenzimidazole is the lower axial ligand of vitamin B12, i.e. cobalamin
-
?
Organism
Organism
UniProt
Commentary
Textmining
Sinorhizobium meliloti
Q92PC8
gene bluB
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
FMNH2 + NADH + H+ + O2
-
728760
Sinorhizobium meliloti
5,6-dimethylbenzimidazole + D-erythrose 4-phosphate + NAD+ + ?
5,6-dimethylbenzimidazole is the lower axial ligand of vitamin B12, i.e. cobalamin
-
-
?
FMNH2 + NADH + H+ + O2
BluB fragments a flavin isoalloxazine ring
728760
Sinorhizobium meliloti
5,6-dimethylbenzimidazole + D-erythrose 4-phosphate + NAD+ + ?
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
BluB
-
Sinorhizobium meliloti
flavin destructase
-
Sinorhizobium meliloti
flavin destructase enzyme
-
Sinorhizobium meliloti
Cofactor
Cofactor
Commentary
Organism
Structure
NADH
-
Sinorhizobium meliloti
Cloned(Commentary) (protein specific)
Commentary
Organism
gene bluB, genotyping
Sinorhizobium meliloti
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADH
-
Sinorhizobium meliloti
Engineering (protein specific)
Protein Variants
Commentary
Organism
A156V
site-directed mutagenesis, inactive mutant
Sinorhizobium meliloti
D32N
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Sinorhizobium meliloti
D32N/S167G
site-directed mutagenesis, the mutant shows no activity
Sinorhizobium meliloti
E78K
site-directed mutagenesis, the mutant is not soluble
Sinorhizobium meliloti
G110S
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
G133S
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
G193D
site-directed mutagenesis, inactive mutant
Sinorhizobium meliloti
G61D
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
M140I
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Sinorhizobium meliloti
M94I
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Sinorhizobium meliloti
P202L
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
P65L
site-directed mutagenesis, inactive mutant
Sinorhizobium meliloti
R30C
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
R30H
site-directed mutagenesis, the mutant shows no 5,6-dimethylbenzimidazole forming activity
Sinorhizobium meliloti
S167G
site-directed mutagenesis, the mutant shows higly reduced activity compared to the wild-type enzyme
Sinorhizobium meliloti
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
FMN/FMNH2 dissociation constants and free energy of binding of wild-type and mutant enzymes, overview
Sinorhizobium meliloti
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
FMNH2 + NADH + H+ + O2
Sinorhizobium meliloti
-
5,6-dimethylbenzimidazole + D-erythrose 4-phosphate + NAD+ + ?
5,6-dimethylbenzimidazole is the lower axial ligand of vitamin B12, i.e. cobalamin
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
FMNH2 + NADH + H+ + O2
-
728760
Sinorhizobium meliloti
5,6-dimethylbenzimidazole + D-erythrose 4-phosphate + NAD+ + ?
5,6-dimethylbenzimidazole is the lower axial ligand of vitamin B12, i.e. cobalamin
-
-
?
FMNH2 + NADH + H+ + O2
BluB fragments a flavin isoalloxazine ring
728760
Sinorhizobium meliloti
5,6-dimethylbenzimidazole + D-erythrose 4-phosphate + NAD+ + ?
-
-
-
?
General Information
General Information
Commentary
Organism
additional information
identification of catalytic residues involved in the reaction, mutational analysis, overview. The enzyme shows interactions with the phosphate group and ribityl tail of FMN
Sinorhizobium meliloti
General Information (protein specific)
General Information
Commentary
Organism
additional information
identification of catalytic residues involved in the reaction, mutational analysis, overview. The enzyme shows interactions with the phosphate group and ribityl tail of FMN
Sinorhizobium meliloti
Other publictions for EC 1.14.99.40
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
728590
Collins
Bacillus megaterium has both a ...
Bacillus megaterium, Bacillus megaterium DSM 319
PLoS ONE
8
e55708
2013
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728760
Yu
Active site residues critical ...
Sinorhizobium meliloti
Protein Sci.
21
839-849
2012
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15
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15
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