BRENDA - Enzyme Database
show all sequences of 1.14.13.44

Structures of the Apo and FAD-bound forms of 2-hydroxybiphenyl 3-monooxygenase (HbpA) locate activity hotspots identified by using directed evolution

Jensen, C.N.; Mielke, T.; Farrugia, J.E.; Frank, A.; Man, H.; Hart, S.; Turkenburg, J.P.; Grogan, G.; ChemBioChem 16, 968-976 (2015)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expression in Escherichia coli BL21
Pseudomonas nitroreducens
Crystallization (Commentary)
Crystallization (Commentary)
Organism
single crystals of the enzyme are used to determine its structure in two forms: an FAD-bound form that allows characterisation of the active site, and an apo form that, although lacking flavin, gives extra information on the location of residues and structure of mobile loops that are absent from the FAD complex
Pseudomonas nitroreducens
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0031
-
2-Hydroxybiphenyl
pH 7.5, temperature not specified in the publication, mutant enzyme H48A
Pseudomonas nitroreducens
0.0033
-
2-Hydroxybiphenyl
pH 7.5, temperature not specified in the publication, wild-type enzyme
Pseudomonas nitroreducens
0.0048
-
2-Hydroxybiphenyl
pH 7.5, temperature not specified in the publication R242A
Pseudomonas nitroreducens
0.005
-
2-Hydroxybiphenyl
pH 7.5, temperature not specified in the publication, mutant enzyme D117A
Pseudomonas nitroreducens
Organism
Organism
UniProt
Commentary
Textmining
Pseudomonas nitroreducens
O06647
-
-
Purification (Commentary)
Purification (Commentary)
Organism
-
Pseudomonas nitroreducens
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2-hydroxybiphenyl + NADH + H+ + O2
-
744702
Pseudomonas nitroreducens
2,3-dihydroxybiphenyl + NAD+ + H2O
-
-
-
?
Subunits
Subunits
Commentary
Organism
tetramer
the structure of the tetramer is determined, there is a large cavity at the centre of the tetramer into which the side chains of several hydrophilic amino acids of each monomer, including Glu75 and Tyr76, are projected
Pseudomonas nitroreducens
Synonyms
Synonyms
Commentary
Organism
HbpA
-
Pseudomonas nitroreducens
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Pseudomonas nitroreducens
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
FAD-dependent monooxygenase
Pseudomonas nitroreducens
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli BL21
Pseudomonas nitroreducens
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
FAD-dependent monooxygenase
Pseudomonas nitroreducens
Crystallization (Commentary) (protein specific)
Crystallization
Organism
single crystals of the enzyme are used to determine its structure in two forms: an FAD-bound form that allows characterisation of the active site, and an apo form that, although lacking flavin, gives extra information on the location of residues and structure of mobile loops that are absent from the FAD complex
Pseudomonas nitroreducens
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0031
-
2-Hydroxybiphenyl
pH 7.5, temperature not specified in the publication, mutant enzyme H48A
Pseudomonas nitroreducens
0.0033
-
2-Hydroxybiphenyl
pH 7.5, temperature not specified in the publication, wild-type enzyme
Pseudomonas nitroreducens
0.0048
-
2-Hydroxybiphenyl
pH 7.5, temperature not specified in the publication R242A
Pseudomonas nitroreducens
0.005
-
2-Hydroxybiphenyl
pH 7.5, temperature not specified in the publication, mutant enzyme D117A
Pseudomonas nitroreducens
Purification (Commentary) (protein specific)
Commentary
Organism
-
Pseudomonas nitroreducens
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2-hydroxybiphenyl + NADH + H+ + O2
-
744702
Pseudomonas nitroreducens
2,3-dihydroxybiphenyl + NAD+ + H2O
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer
the structure of the tetramer is determined, there is a large cavity at the centre of the tetramer into which the side chains of several hydrophilic amino acids of each monomer, including Glu75 and Tyr76, are projected
Pseudomonas nitroreducens
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Pseudomonas nitroreducens
Other publictions for EC 1.14.13.44
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)
744407
Kanteev
A crystal structure of 2-hydr ...
Pseudomonas nitroreducens
Biochim. Biophys. Acta
1854
1906-1913
2015
-
-
1
1
6
-
-
12
-
-
-
-
-
5
-
-
1
-
-
-
-
-
2
-
2
-
-
-
12
-
-
-
1
-
-
-
-
-
1
1
1
6
-
-
-
-
12
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
12
-
-
-
-
-
-
-
-
12
12
744702
Jensen
Structures of the Apo and FAD ...
Pseudomonas nitroreducens
ChemBioChem
16
968-976
2015
-
-
1
1
-
-
-
4
-
-
-
-
-
3
-
-
1
-
-
-
-
-
1
1
1
-
-
-
-
1
-
-
1
-
-
-
-
-
1
1
1
-
-
-
-
-
4
-
-
-
-
-
-
-
1
-
-
-
-
1
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
657455
Meyer
Crystallization and preliminar ...
Pseudomonas nitroreducens
Acta Crystallogr. Sect. D
59
741-743
2003
-
-
1
1
-
-
3
-
-
-
1
1
-
7
-
-
1
-
-
-
-
-
2
1
1
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
1
-
-
-
3
-
-
-
-
1
1
-
-
-
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
438847
Meyer
Changing the substrate reactiv ...
Pseudomonas nitroreducens, Pseudomonas nitroreducens HBP1
J. Biol. Chem.
277
5575-5582
2002
-
-
-
-
2
-
-
1
-
-
-
-
-
10
-
-
-
-
-
-
-
-
10
-
-
-
-
-
4
-
-
-
2
-
-
-
-
-
-
2
-
2
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
10
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
438848
Meyer
Hydroxylation of indole by lab ...
Pseudomonas nitroreducens, Pseudomonas nitroreducens HBP1
J. Biol. Chem.
277
34161-34167
2002
-
-
-
-
1
-
-
-
-
-
-
-
-
10
-
-
-
-
-
-
-
-
6
-
-
-
-
-
4
-
-
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
438849
Suske
Catalytic mechanism of 2-hydro ...
Pseudomonas nitroreducens, Pseudomonas nitroreducens HBP1
J. Biol. Chem.
274
33355-33365
1999
-
-
-
-
-
-
-
2
-
-
-
-
-
10
-
-
1
1
-
-
-
-
8
-
-
-
-
-
1
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
-
1
-
-
-
-
8
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
658323
Held
An integrated process for the ...
Escherichia coli, Escherichia coli JM101
Biotechnol. Bioeng.
62
641-648
1999
-
1
-
-
-
-
-
-
-
-
-
-
-
12
-
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
1
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
438850
Suske
Purification and characterizat ...
Pseudomonas nitroreducens, Pseudomonas nitroreducens HBP1
J. Biol. Chem.
272
24257-24265
1997
-
-
-
-
-
-
7
10
-
-
2
2
-
16
-
-
1
-
-
-
1
1
18
1
-
-
-
-
11
1
1
-
3
1
-
-
-
-
-
3
-
-
-
-
7
1
10
-
-
2
2
-
-
-
1
-
-
1
1
18
1
-
-
-
11
1
1
-
-
-
-
-
-
-
-