BRENDA - Enzyme Database
show all sequences of 1.14.15.8

Theoretical and experimental evaluation of a CYP106A2 low homology model and production of mutants with changed activity and selectivity of hydroxylation

Lisurek, M.; Simgen, B.; Antes, I.; Bernhardt, R.; Chembiochem 16, 1439-1449 (2008)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expression in Escherichia coli
Bacillus megaterium
Engineering
Protein Variants
Commentary
Organism
A395L
Vmax/Km for progesterone is 2fold lower than wild-type value
Bacillus megaterium
E90V/D185G
Vmax/Km for progesterone is 1.3fold higher than wild-type value
Bacillus megaterium
G397P
the mutant exhibits 2% of the wild-type activity, Vmax/Km for progesterone is 51.7fold lower than wild-type value
Bacillus megaterium
I86T
Vmax/Km for progesterone is 2fold higher than wild-type value
Bacillus megaterium
K27R/I71T/I215T
Vmax/Km for progesterone is 2.5fold higher than wild-type value
Bacillus megaterium
Q398S
Vmax/Km for progesterone is 5fold lower than wild-type value
Bacillus megaterium
S394I
Vmax/Km for progesterone is 7.5fold lower than wild-type value
Bacillus megaterium
T396R
mutant does not produce any hydroxylated product up to an adrenodoxin concentration of 0.1 mM
Bacillus megaterium
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0007
-
progesterone
pH 7.4, 30C, mutant K27R/I71T/I215T
Bacillus megaterium
0.0009
-
progesterone
pH 7.4, 30C, mutant I86T
Bacillus megaterium
0.001
-
progesterone
pH 7.4, 30C, mutant E90V/D185G
Bacillus megaterium
0.0012
-
deoxycorticosterone
pH 7.4, 30C, mutant I86T
Bacillus megaterium
0.0015
-
deoxycorticosterone
pH 7.4, 30C, wild-type enzyme
Bacillus megaterium
0.0015
-
progesterone
pH 7.4, 30C, wild-type enzyme
Bacillus megaterium
0.0019
-
deoxycorticosterone
pH 7.4, 30C, mutant E90V/D185G
Bacillus megaterium
0.0024
-
deoxycorticosterone
pH 7.4, 30C, mutant K27R/I71T/I215T
Bacillus megaterium
0.0024
-
progesterone
pH 7.4, 30C, mutant A395L
Bacillus megaterium
0.0042
-
deoxycorticosterone
pH 7.4, 30C, mutant A395L
Bacillus megaterium
0.0043
-
progesterone
pH 7.4, 30C, mutant S394I
Bacillus megaterium
0.0046
-
deoxycorticosterone
pH 7.4, 30C, mutant G397P
Bacillus megaterium
0.0047
-
deoxycorticosterone
pH 7.4, 30C, mutant S394I
Bacillus megaterium
0.0049
-
progesterone
pH 7.4, 30C, mutant Q398S
Bacillus megaterium
0.005
-
progesterone
pH 7.4, 30C, mutant G397P
Bacillus megaterium
0.0056
-
deoxycorticosterone
pH 7.4, 30C, mutant Q398S
Bacillus megaterium
Organism
Organism
UniProt
Commentary
Textmining
Bacillus megaterium
Q06069
ATCC 13368
-
Purification (Commentary)
Purification (Commentary)
Organism
-
Bacillus megaterium
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
11-deoxycorticosterone + reduced adrenodoxin + O2
adrenodoxin is a [2Fe-2S] ferredoxin involved in electron transfer from NADPH+ (via a reductase) to cytochrome P-450 in the adrenal gland, 15beta-hydroxy-deoxycorticosterone is the only product that is observed, both in the case of wild-type CYP106A2 and all the mutants (except T396R)
697323
Bacillus megaterium
15beta-hydroxy-11-deoxycorticosterone + oxidized adrenodoxin + H2O
-
-
-
?
progesterone + reduced adrenodoxin + O2
adrenodoxin is a [2Fe-2S] ferredoxin involved in electron transfer from NADPH+ (via a reductase) to cytochrome P-450 in the adrenal gland, 15beta-hydroxy-progesterone is the main product of wild-type enzyme and all mutants. In order to gain insights into the structure and function of CYP106A2, whose crystal structure is unknown, a homology model has been created. The substrate progesterone is then docked into the active site to predict which residues might affect substrate binding. The model is substantiated by using a combination of theoretical and experimental investigations
697323
Bacillus megaterium
15beta-hydroxy-progesterone + oxidized adrenodoxin + H2O
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
CYP106A2
-
Bacillus megaterium
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
30
-
assay at
Bacillus megaterium
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Bacillus megaterium
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli
Bacillus megaterium
Engineering (protein specific)
Protein Variants
Commentary
Organism
A395L
Vmax/Km for progesterone is 2fold lower than wild-type value
Bacillus megaterium
E90V/D185G
Vmax/Km for progesterone is 1.3fold higher than wild-type value
Bacillus megaterium
G397P
the mutant exhibits 2% of the wild-type activity, Vmax/Km for progesterone is 51.7fold lower than wild-type value
Bacillus megaterium
I86T
Vmax/Km for progesterone is 2fold higher than wild-type value
Bacillus megaterium
K27R/I71T/I215T
Vmax/Km for progesterone is 2.5fold higher than wild-type value
Bacillus megaterium
Q398S
Vmax/Km for progesterone is 5fold lower than wild-type value
Bacillus megaterium
S394I
Vmax/Km for progesterone is 7.5fold lower than wild-type value
Bacillus megaterium
T396R
mutant does not produce any hydroxylated product up to an adrenodoxin concentration of 0.1 mM
Bacillus megaterium
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0007
-
progesterone
pH 7.4, 30C, mutant K27R/I71T/I215T
Bacillus megaterium
0.0009
-
progesterone
pH 7.4, 30C, mutant I86T
Bacillus megaterium
0.001
-
progesterone
pH 7.4, 30C, mutant E90V/D185G
Bacillus megaterium
0.0012
-
deoxycorticosterone
pH 7.4, 30C, mutant I86T
Bacillus megaterium
0.0015
-
deoxycorticosterone
pH 7.4, 30C, wild-type enzyme
Bacillus megaterium
0.0015
-
progesterone
pH 7.4, 30C, wild-type enzyme
Bacillus megaterium
0.0019
-
deoxycorticosterone
pH 7.4, 30C, mutant E90V/D185G
Bacillus megaterium
0.0024
-
deoxycorticosterone
pH 7.4, 30C, mutant K27R/I71T/I215T
Bacillus megaterium
0.0024
-
progesterone
pH 7.4, 30C, mutant A395L
Bacillus megaterium
0.0042
-
deoxycorticosterone
pH 7.4, 30C, mutant A395L
Bacillus megaterium
0.0043
-
progesterone
pH 7.4, 30C, mutant S394I
Bacillus megaterium
0.0046
-
deoxycorticosterone
pH 7.4, 30C, mutant G397P
Bacillus megaterium
0.0047
-
deoxycorticosterone
pH 7.4, 30C, mutant S394I
Bacillus megaterium
0.0049
-
progesterone
pH 7.4, 30C, mutant Q398S
Bacillus megaterium
0.005
-
progesterone
pH 7.4, 30C, mutant G397P
Bacillus megaterium
0.0056
-
deoxycorticosterone
pH 7.4, 30C, mutant Q398S
Bacillus megaterium
Purification (Commentary) (protein specific)
Commentary
Organism
-
Bacillus megaterium
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
11-deoxycorticosterone + reduced adrenodoxin + O2
adrenodoxin is a [2Fe-2S] ferredoxin involved in electron transfer from NADPH+ (via a reductase) to cytochrome P-450 in the adrenal gland, 15beta-hydroxy-deoxycorticosterone is the only product that is observed, both in the case of wild-type CYP106A2 and all the mutants (except T396R)
697323
Bacillus megaterium
15beta-hydroxy-11-deoxycorticosterone + oxidized adrenodoxin + H2O
-
-
-
?
progesterone + reduced adrenodoxin + O2
adrenodoxin is a [2Fe-2S] ferredoxin involved in electron transfer from NADPH+ (via a reductase) to cytochrome P-450 in the adrenal gland, 15beta-hydroxy-progesterone is the main product of wild-type enzyme and all mutants. In order to gain insights into the structure and function of CYP106A2, whose crystal structure is unknown, a homology model has been created. The substrate progesterone is then docked into the active site to predict which residues might affect substrate binding. The model is substantiated by using a combination of theoretical and experimental investigations
697323
Bacillus megaterium
15beta-hydroxy-progesterone + oxidized adrenodoxin + H2O
-
-
-
?
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
30
-
assay at
Bacillus megaterium
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Bacillus megaterium
Other publictions for EC
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)
745540
Kim
Crystal structure and functio ...
Bacillus sp. PAMC 23377
J. Microbiol. Biotechnol.
27
1472-1482
2017
-
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1
1
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3
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2
1
1
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1
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1
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2
1
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-
-
-
-
-
-
-
-
-
746507
Nikolaus
Engineering of CYP106A2 for s ...
Bacillus megaterium
Steroids
120
41-48
2017
-
-
-
-
2
-
-
-
-
-
-
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1
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744706
Janocha
Crystal structure of CYP106A2 ...
Bacillus megaterium
ChemBioChem
17
852-860
2016
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1
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3
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744148
Kiss
Comparison of CYP106A1 and CY ...
Bacillus megaterium
Appl. Microbiol. Biotechnol.
99
8495-8514
2015
-
-
1
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-
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4
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24
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2
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1
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2
2
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24
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745699
Schmitz
Steroid conversion with CYP10 ...
Bacillus megaterium
Microb. Cell Fact.
13
81
2014
-
1
1
-
-
-
-
-
-
-
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1
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6
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6
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724061
Bleif
A new Bacillus megaterium whol ...
Bacillus megaterium, Bacillus megaterium ATCC 13368
Appl. Microbiol. Biotechnol.
93
1135-1146
2012
-
-
1
-
1
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1
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2
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6
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1
1
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1
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2
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1
2
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1
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6
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1
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1
-
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725584
Schumacher
Autodisplay of functional CYP1 ...
Bacillus megaterium
J. Biotechnol.
161
104-112
2012
-
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1
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1
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1
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4
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1
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1
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1
3
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1
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4
-
1
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1
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711747
Bleif
Identification of CYP106A2 as ...
Bacillus megaterium, Bacillus megaterium ATCC 13368
ChemBioChem
12
576-582
2011
-
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1
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2
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4
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697322
Zehentgruber
Towards preparative scale ster ...
Bacillus megaterium
Chembiochem
11
713-721
2010
-
1
1
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1
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696462
Goni
Cyanobacterial electron carrie ...
Bacillus megaterium
Biochim. Biophys. Acta
1794
1635-1642
2009
-
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3
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6
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1
1
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3
1
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2
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1
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3
1
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693956
Virus
Molecular evolution of a stero ...
Bacillus megaterium
Lipids
43
1133-1141
2008
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1
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6
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5
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1
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1
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697323
Lisurek
Theoretical and experimental e ...
Bacillus megaterium
Chembiochem
16
1439-1449
2008
-
-
1
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8
-
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16
-
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1
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2
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1
1
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8
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16
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2
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1
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699068
Ruijssenaars
Testosterone 15beta-hydroxylat ...
Bacillus megaterium
J. Biotechnol.
131
205-208
2007
-
1
1
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1
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1
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696142
Virus
Function and engineering of th ...
Bacillus megaterium
Biochem. Soc. Trans.
34
1215-1218
2006
-
-
1
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2
1
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1
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3
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3
1
2
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1
1
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2
1
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1
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1
3
1
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695981
Lisurek
Identification of monohydroxy ...
Bacillus megaterium
Biochem. Biophys. Res. Commun.
319
677-682
2004
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1
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1
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1
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1
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695980
Simgen
Substrate binding to 15beta-hy ...
Bacillus megaterium
Biochem. Biophys. Res. Commun.
269
737-742
2000
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1
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700219
Rauschenbach
Cloning sequencing and express ...
Bacillus megaterium
Mol. Gen. Genet.
241
170-176
1993
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1
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6
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1
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2
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698684
Berg
Purification and characterizat ...
Bacillus megaterium
J. Biol. Chem.
254
5264-5271
1979
1
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1
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1
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698682
Berg
Characterization of a cytochro ...
Bacillus megaterium
J. Biol. Chem.
251
2831-2838
1976
2
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8
7
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4
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8
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1
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8
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8
8
7
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8
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