BRENDA - Enzyme Database show
show all sequences of 1.1.1.159

Roles of the Ser146, Tyr159, and Lys163 residues in the catalytic action of 7alpha-hydroxysteroid dehydrogenase from Escherichia coli

Tanabe, T.; Tanaka, N.; Uchikawa, K.; Kabashima, T.; Ito, K.; Nonaka, T.; Mitsui, Y.; Tsuru, M.; Yoshimoto, T.; J. Biochem. 124, 634-641 (1998)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
site directed mutagenesis
Escherichia coli
Crystallization (Commentary)
Crystallization
Organism
-
Escherichia coli
Engineering
Amino acid exchange
Commentary
Organism
K163I
5.25% activity of wild-type activity
Escherichia coli
K163R
63.7% activity of wild-type activity
Escherichia coli
S146A
20.3% activity of wild-type activity
Escherichia coli
S146H
35.6% activity of wild-type activity
Escherichia coli
Y159F
no activity
Escherichia coli
Y159H
13.3% activity of wild-type activity
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.159
-
NAD+
mutant K163R
Escherichia coli
0.223
-
cholic acid
mutant K163I
Escherichia coli
0.279
-
NAD+
-
Escherichia coli
0.283
-
cholic acid
mutant S146H
Escherichia coli
0.302
-
NAD+
mutant S146A
Escherichia coli
0.329
-
NAD+
mutant K163I
Escherichia coli
0.361
-
cholic acid
-
Escherichia coli
0.364
-
NAD+
mutant S146H
Escherichia coli
0.386
-
NAD+
mutant Y159H
Escherichia coli
0.407
-
cholic acid
mutant Y159H
Escherichia coli
0.415
-
cholic acid
mutant S146A
Escherichia coli
0.612
-
cholic acid
mutant K163R
Escherichia coli
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
28000
-
4 * 28000, SDS-PAGE
Escherichia coli
107100
-
sequence determination
Escherichia coli
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
-
-
-
Purification (Commentary)
Commentary
Organism
-
Escherichia coli
Reaction
Reaction
Commentary
Organism
cholate + NAD+ = 3alpha,12alpha-dihydroxy-7-oxo-5beta-cholan-24-oate + NADH + H+
Tyr159 acts as basic catalyst, Lys163 binds to NAD(H) and lowers the pKa value of Tyr159, Ser146 stabilizes the substrate, reaction intermediate and product in catalysis
Escherichia coli
Subunits
Subunits
Commentary
Organism
tetramer
4 * 28000, SDS-PAGE
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
7.92
-
cholic acid
mutant K163I
Escherichia coli
20.1
-
cholic acid
mutant Y159H
Escherichia coli
30.7
-
cholic acid
mutant S146A
Escherichia coli
53.7
-
cholic acid
mutant S146H
Escherichia coli
96.2
-
cholic acid
mutant K163R
Escherichia coli
151
-
cholic acid
-
Escherichia coli
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
10
-
oxidation of cholic acid
Escherichia coli
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6
10
-
Escherichia coli
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
wild type and mutants
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
site directed mutagenesis
Escherichia coli
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
wild type and mutants
Escherichia coli
Crystallization (Commentary) (protein specific)
Crystallization
Organism
-
Escherichia coli
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
K163I
5.25% activity of wild-type activity
Escherichia coli
K163R
63.7% activity of wild-type activity
Escherichia coli
S146A
20.3% activity of wild-type activity
Escherichia coli
S146H
35.6% activity of wild-type activity
Escherichia coli
Y159F
no activity
Escherichia coli
Y159H
13.3% activity of wild-type activity
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.159
-
NAD+
mutant K163R
Escherichia coli
0.223
-
cholic acid
mutant K163I
Escherichia coli
0.279
-
NAD+
-
Escherichia coli
0.283
-
cholic acid
mutant S146H
Escherichia coli
0.302
-
NAD+
mutant S146A
Escherichia coli
0.329
-
NAD+
mutant K163I
Escherichia coli
0.361
-
cholic acid
-
Escherichia coli
0.364
-
NAD+
mutant S146H
Escherichia coli
0.386
-
NAD+
mutant Y159H
Escherichia coli
0.407
-
cholic acid
mutant Y159H
Escherichia coli
0.415
-
cholic acid
mutant S146A
Escherichia coli
0.612
-
cholic acid
mutant K163R
Escherichia coli
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
28000
-
4 * 28000, SDS-PAGE
Escherichia coli
107100
-
sequence determination
Escherichia coli
Purification (Commentary) (protein specific)
Commentary
Organism
-
Escherichia coli
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer
4 * 28000, SDS-PAGE
Escherichia coli
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
7.92
-
cholic acid
mutant K163I
Escherichia coli
20.1
-
cholic acid
mutant Y159H
Escherichia coli
30.7
-
cholic acid
mutant S146A
Escherichia coli
53.7
-
cholic acid
mutant S146H
Escherichia coli
96.2
-
cholic acid
mutant K163R
Escherichia coli
151
-
cholic acid
-
Escherichia coli
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
10
-
oxidation of cholic acid
Escherichia coli
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6
10
-
Escherichia coli
Other publictions for EC 1.1.1.159
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
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)
697994
Fukiya
Conversion of cholic acid and ...
Bacteroides fragilis, Bacteroides fragilis NCTC 9343, Bacteroides intestinalis, Bacteroides intestinalis am-1, Escherichia coli HB101
FEMS Microbiol. Lett.
293
263-270
2009
6
-
-
-
-
-
-
-
-
-
-
-
-
13
-
-
-
-
-
-
-
-
10
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
10
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
701240
Giovannini
7alpha- and 12alpha-Hydroxyste ...
Acinetobacter lwoffii
Steroids
73
1385-1390
2008
1
1
-
-
-
-
-
-
-
-
-
-
-
2
-
-
1
-
-
-
2
1
1
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
2
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
670898
Pedrini
Xanthomonas maltophilia CBS 89 ...
Stenotrophomonas maltophilia, Stenotrophomonas maltophilia CBS 897.97
Steroids
71
189-198
2006
-
1
-
-
-
-
-
2
-
-
-
1
-
3
-
-
1
-
-
-
2
1
7
-
1
-
-
-
1
-
-
2
-
-
-
-
1
-
2
-
-
-
-
-
-
2
-
-
-
1
-
-
-
1
-
-
2
1
7
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
670926
Zhu
-
Enzymatic enantioselective red ...
Bacteroides fragilis
Tetrahedron
62
4535-4539
2006
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
1
-
-
-
1
-
16
-
1
-
1
-
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
1
-
16
-
1
-
1
-
1
-
-
-
-
-
-
-
-
-
656301
Ueda
-
Molecular and enzymatic proper ...
Pseudomonas sp., Pseudomonas sp. B-0831
J. Biol. Macromol.
4
33-38
2004
-
-
-
-
-
-
-
1
-
-
2
-
-
6
-
-
1
-
-
-
-
-
6
1
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
1
-
-
2
-
-
-
-
1
-
-
-
-
6
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
655302
Bennett
Cloning and characterization o ...
Bacteroides fragilis, Clostridium scindens, Escherichia coli, Paeniclostridium sordellii
Curr. Microbiol.
47
475-484
2003
-
-
1
-
-
-
2
4
-
-
2
-
-
6
-
-
1
-
-
-
1
-
8
1
-
-
4
-
2
1
-
5
-
-
-
-
-
1
5
-
-
-
-
2
-
4
-
-
2
-
-
-
-
1
-
-
1
-
8
1
-
-
4
-
2
1
-
-
-
-
-
-
-
-
286083
Tanabe
Roles of the Ser146, Tyr159, a ...
Escherichia coli
J. Biochem.
124
634-641
1998
-
-
1
1
6
-
-
12
-
-
2
-
-
2
-
-
1
1
-
-
-
-
-
1
-
-
-
6
1
1
-
1
-
-
-
-
-
1
1
1
6
-
-
-
-
12
-
-
2
-
-
-
-
1
-
-
-
-
-
1
-
-
-
6
1
1
-
-
-
-
-
-
-
-
286081
Tanaka
Crystal structures of the bina ...
Escherichia coli
Biochemistry
35
7715-7730
1996
-
-
-
1
-
-
-
-
-
-
-
1
-
2
-
-
1
1
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286084
Coleman
Characterization and regulatio ...
Paeniclostridium sordellii
J. Bacteriol.
176
4865-4874
1994
-
-
1
-
-
-
-
-
-
-
1
1
-
1
-
-
1
1
-
-
1
-
2
1
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
-
1
1
-
-
-
1
-
-
1
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286082
Baron
Cloning, sequencing, and expre ...
Eubacterium sp., Eubacterium sp. VPI 12708
J. Bacteriol.
173
4558-4569
1991
-
-
1
-
-
-
-
-
-
-
1
4
-
12
-
-
1
-
-
-
2
-
8
1
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
-
1
4
-
-
-
1
-
-
2
-
8
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286085
Yoshimoto
Cloning and sequencing of the ...
Escherichia coli
J. Bacteriol.
173
2173-2179
1991
-
-
1
1
-
-
12
5
-
-
2
-
-
3
-
-
1
-
-
-
-
-
6
1
-
-
-
5
1
-
1
1
-
-
-
-
-
1
1
1
-
-
-
12
-
5
-
-
2
-
-
-
-
1
-
-
-
-
6
1
-
-
-
5
1
-
1
-
-
-
-
-
-
-
286072
Prabha
Purification of 7alpha-hydroxy ...
Escherichia coli
Can. J. Microbiol.
36
131-135
1990
-
-
-
-
-
-
-
-
-
-
1
-
-
2
-
-
1
-
-
-
1
-
-
-
-
-
2
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
1
-
-
-
-
-
2
-
1
-
-
-
-
-
-
-
-
-
286073
Prabha
Kinetic properties of 7alpha-h ...
Escherichia coli
Can. J. Microbiol.
35
1076-1080
1989
2
-
-
-
-
-
18
-
-
4
-
-
-
1
-
-
-
-
-
-
8
-
6
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
18
-
-
-
4
-
-
-
-
-
-
-
-
8
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286074
Ottolina
Enzymatic synthesis of [4R-2H] ...
Escherichia coli
Biochim. Biophys. Acta
998
173-178
1989
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286075
Kinoshita
-
Purification and properties of ...
Brevibacterium fuscum
J. Ferment. Technol.
66
145-152
1988
-
-
-
-
-
-
-
8
-
-
2
-
-
1
-
-
1
-
-
-
4
-
8
1
1
1
2
-
3
-
2
1
-
-
-
-
-
-
1
-
-
-
-
-
-
8
-
-
2
-
-
-
-
1
-
-
4
-
8
1
1
1
2
-
3
-
2
-
-
-
-
-
-
-
286076
MacDonald
Bile induction of 7alpha- and ...
Clostridium sardiniense
Biochim. Biophys. Acta
665
262-269
1981
-
-
-
-
-
-
-
3
-
-
-
1
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
286077
Haslewood
The specificity of a 7alpha-hy ...
Escherichia coli
Biochem. J.
157
207-210
1976
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286078
Hylemon
Multiple forms of 7-alpha-hydr ...
Bacteroides fragilis
J. Bacteriol.
122
418-424
1975
-
-
-
-
-
-
2
-
-
2
2
1
-
1
-
-
1
-
-
-
-
-
7
-
-
-
2
-
1
-
-
2
-
-
-
-
-
-
2
-
-
-
-
2
-
-
-
2
2
1
-
-
-
1
-
-
-
-
7
-
-
-
2
-
1
-
-
-
-
-
-
-
-
-
286079
MacDonald
NAD- and NADP-dependent 7alpha ...
Bacteroides fragilis
Biochim. Biophys. Acta
384
12-24
1975
-
-
-
-
-
1
-
7
-
-
-
-
-
3
-
-
-
-
-
-
-
-
5
-
-
-
-
-
2
-
-
2
-
-
-
-
-
-
2
-
-
1
-
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
286080
MacDonald
7alpha-hydroxysteroid dehydrog ...
Escherichia coli
Biochim. Biophys. Acta
309
243-253
1973
-
-
-
-
-
2
3
6
-
-
-
1
-
2
-
-
-
-
-
-
-
-
7
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
1
-
-
2
-
3
-
6
-
-
-
1
-
-
-
-
-
-
-
-
7
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-