BRENDA - Enzyme Database
show all sequences of 1.14.15.30

Characterization and engineering of 3-ketosteroid-Delta1-dehydrogenase and 3-ketosteroid-9alpha-hydroxylase in Mycobacterium neoaurum ATCC 25795 to produce 9alpha-hydroxy-4-androstene-3,17-dione through the catabolism of sterols

Yao, K.; Xu, L.Q.; Wang, F.Q.; Wei, D.Z.; Metab. Eng. 24, 181-191 (2014)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli BL21(DE3) cells
Mycolicibacterium neoaurum
Engineering
Amino acid exchange
Commentary
Organism
additional information
recombinant coexpression of the enzyme KshA from Mycobacterium neoaurum strain NwIB-01 and the three homologues of ketosteroid-DELTA1-dehydrogenase (KstD) from Mycobacterium neoaurum ATCC257953 in Escherichia coli strain BL21(DE3)
Mycolicibacterium neoaurum
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
a Rieske 2Fe-2S enzyme
Mycolicibacterium neoaurum
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycolicibacterium neoaurum
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycolicibacterium neoaurum NwIB-01
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycolicibacterium neoaurum ATCC 25795
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mycolicibacterium neoaurum
C9E9N6
-
-
Mycolicibacterium neoaurum ATCC 25795
C9E9N6
-
-
Mycolicibacterium neoaurum NwIB-01
C9E9N6
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
743221
Mycolicibacterium neoaurum
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
743221
Mycolicibacterium neoaurum NwIB-01
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
743221
Mycolicibacterium neoaurum ATCC 25795
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Mycolicibacterium neoaurum
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Mycolicibacterium neoaurum
Cofactor
Cofactor
Commentary
Organism
Structure
[2Fe-2S]-center
a Rieske 2Fe-2S enzyme
Mycolicibacterium neoaurum
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli BL21(DE3) cells
Mycolicibacterium neoaurum
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
[2Fe-2S]-center
a Rieske 2Fe-2S enzyme
Mycolicibacterium neoaurum
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
recombinant coexpression of the enzyme KshA from Mycobacterium neoaurum strain NwIB-01 and the three homologues of ketosteroid-DELTA1-dehydrogenase (KstD) from Mycobacterium neoaurum ATCC257953 in Escherichia coli strain BL21(DE3)
Mycolicibacterium neoaurum
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
a Rieske 2Fe-2S enzyme
Mycolicibacterium neoaurum
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycolicibacterium neoaurum
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycolicibacterium neoaurum NwIB-01
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycolicibacterium neoaurum ATCC 25795
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
743221
Mycolicibacterium neoaurum
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
743221
Mycolicibacterium neoaurum NwIB-01
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
743221
Mycolicibacterium neoaurum ATCC 25795
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Mycolicibacterium neoaurum
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Mycolicibacterium neoaurum
Expression
Organism
Commentary
Expression
Mycolicibacterium neoaurum
recombinant coexpression of the enzyme KshA from Mycobacterium neoaurum strain NwIB-01 and the three homologues of ketosteroid-DELTA1-dehydrogenase (KstD) from Mycobacterium neoaurum ATCC257953 in Escherichia coli strain BL21(DE3),to install a steroid production system with a broad substrate specificity. Enzyme KshA converts the KstD reaction side product 4-androstene-3,17-dione into 9alpha-hydroxyandrosta-1,4-diene-3,17-dione
additional information
Expression (protein specific)
Organism
Commentary
Expression
Mycolicibacterium neoaurum
recombinant coexpression of the enzyme KshA from Mycobacterium neoaurum strain NwIB-01 and the three homologues of ketosteroid-DELTA1-dehydrogenase (KstD) from Mycobacterium neoaurum ATCC257953 in Escherichia coli strain BL21(DE3),to install a steroid production system with a broad substrate specificity. Enzyme KshA converts the KstD reaction side product 4-androstene-3,17-dione into 9alpha-hydroxyandrosta-1,4-diene-3,17-dione
additional information
Other publictions for EC 1.14.15.30
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)
745648
Guevara
Functional characterization o ...
Rhodococcus ruber, Rhodococcus ruber Chol-4
J. Steroid Biochem. Mol. Biol.
172
176-187
2017
-
-
1
-
1
-
-
-
-
-
-
4
-
6
-
-
-
-
-
-
-
-
22
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
22
1
-
-
-
-
-
-
-
-
-
4
4
-
-
-
746503
Zhang
Efficient 9alpha-hydroxy-4-an ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum JC-12
SpringerPlus
5
1207
2016
-
-
1
-
1
-
5
-
-
-
-
8
-
2
-
-
1
-
-
-
1
1
10
2
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
5
-
-
-
-
-
8
-
-
-
1
-
-
1
1
10
2
1
-
-
-
1
-
-
-
-
1
1
-
-
-
743221
Yao
Characterization and engineer ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum ATCC 25795, Mycolicibacterium neoaurum NwIB-01
Metab. Eng.
24
181-191
2014
-
-
1
-
1
-
-
-
-
1
-
3
-
4
-
-
-
-
-
-
-
-
3
-
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
-
1
-
-
-
-
-
-
1
-
3
-
-
-
-
-
-
-
-
3
-
1
-
-
-
1
-
-
-
1
-
-
1
-
-
744058
Petrusma
3-Ketosteroid 9alpha-hydroxyl ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv, Mycolicibacterium smegmatis, Mycolicibacterium smegmatis mc2 155, Rhodococcus erythropolis, Rhodococcus erythropolis SQ1, Rhodococcus jostii, Rhodococcus rhodochrous, Rhodococcus rhodochrous DSM 43269
Antonie van Leeuwenhoek
106
157-172
2014
-
2
2
-
1
-
-
-
-
5
-
22
-
15
-
-
-
5
-
-
-
-
28
5
-
-
-
-
-
-
-
10
-
-
-
-
2
4
14
-
1
-
-
-
-
-
-
7
-
22
-
-
-
-
-
-
-
-
28
7
-
-
-
-
-
-
-
-
-
20
28
-
-
-
745308
Penfield
Substrate specificities and c ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv, Rhodococcus rhodochrous, Rhodococcus rhodochrous DSM43269, Rhodococcus rhodochrous DSM 43269
J. Biol. Chem.
289
25523-25536
2014
-
-
1
2
-
-
4
19
-
-
-
8
-
18
-
-
1
2
-
-
-
-
54
2
2
-
-
19
2
-
-
-
3
-
-
-
-
2
-
3
-
-
-
6
3
20
-
-
-
8
-
-
-
2
-
-
-
-
54
3
3
-
-
19
3
-
-
-
-
4
6
-
17
17
745698
Yeh
Deletion of the gene encoding ...
Rhodococcus hoagii 103S, Rhodococcus hoagii
Microb. Cell Fact.
13
130
2014
-
-
1
-
1
-
-
-
-
-
-
4
-
2
-
-
-
-
-
-
-
-
4
-
1
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
4
-
1
-
-
-
-
-
-
-
-
1
1
-
-
-
717737
Petrusma
Structural features in the Ksh ...
Rhodococcus rhodochrous, Rhodococcus rhodochrous DSM 43269
J. Bacteriol.
194
115-121
2012
-
-
1
-
8
-
-
2
-
1
-
3
-
13
-
-
1
-
-
-
-
-
74
-
-
-
-
-
1
-
-
2
-
-
-
-
-
3
4
-
16
-
-
-
-
4
-
2
-
3
-
-
-
3
-
-
-
-
74
-
-
-
-
-
2
-
-
-
-
1
2
-
-
-
717735
Petrusma
Multiplicity of 3-ketosteroid- ...
Rhodococcus rhodochrous, Rhodococcus rhodochrous DSM 43269
J. Bacteriol.
193
3931-3940
2011
-
-
1
-
1
-
-
-
-
-
-
4
-
19
-
-
1
-
-
1
-
-
75
-
-
-
-
-
1
-
-
1
-
-
-
-
-
6
5
-
5
-
-
-
-
-
-
-
-
4
-
-
-
6
-
3
-
-
75
-
-
-
-
-
3
-
-
-
1
3
11
5
-
-
717854
Capyk
Activity of 3-ketosteroid 9alp ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Biol. Chem.
286
40717-40724
2011
-
-
-
-
-
-
-
13
-
-
-
2
-
9
-
-
-
-
-
-
-
-
30
1
1
-
-
11
1
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
21
-
-
-
2
-
-
-
-
-
-
-
-
30
2
2
-
-
19
2
-
-
-
-
2
3
-
13
19
713784
Wei
A new steroid-transforming str ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum NwIB-01
Appl. Biochem. Biotechnol.
162
1446-1456
2010
-
-
-
-
-
-
-
-
-
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1
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3
-
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2
1
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1
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1
-
-
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-
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-
-
2
1
-
-
-
-
-
-
-
1
-
1
1
-
-
-
716250
Hu
3-Ketosteroid 9alpha-hydroxyla ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Mol. Microbiol.
75
107-121
2010
-
-
-
-
-
-
-
-
-
-
-
-
-
160
-
-
-
-
-
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4
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-
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-
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-
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-
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-
-
-
4
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
717500
Fan
Cloning, heterologous expressi ...
Mycobacterium sp., Mycobacterium sp. NwIB-01
Chin. J. Biotechnol.
25
2014-2021
2009
-
-
1
-
-
-
-
-
-
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1
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8
-
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1
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1
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1
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1
-
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1
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-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
717784
Capyk
Characterization of 3-ketoster ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Biol. Chem.
284
9937-9946
2009
-
-
1
1
-
-
-
3
-
-
-
-
-
6
-
-
1
-
-
-
-
-
4
-
-
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-
3
-
-
-
1
-
-
-
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-
1
1
1
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-
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3
-
-
-
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-
-
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1
-
-
-
-
4
-
-
-
-
3
-
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-
-
-
-
-
-
3
3
717119
Van Der Geize
Characterization of a second R ...
Rhodococcus erythropolis, Rhodococcus erythropolis SQ1
Appl. Environ. Microbiol.
74
7197-7203
2008
-
-
-
-
-
-
-
-
-
-
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9
-
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8
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1
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2
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8
-
-
-
-
-
-
-
-
-
1
2
3
1
-
-
717118
Andor
Generation of useful insertion ...
Mycolicibacterium smegmatis, Mycolicibacterium smegmatis mc(2)155 / ATCC 700084
Appl. Environ. Microbiol.
72
6554-6559
2006
-
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
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1
-
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1
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-
-
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-
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-
-
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-
-
-
-
-
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-
-
-
-
-
-
718125
Van Der Geize
Molecular and functional chara ...
Rhodococcus erythropolis, Rhodococcus erythropolis SQ1
Mol. Microbiol.
45
1007-1018
2002
-
-
-
-
1
-
-
-
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1
-
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4
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8
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1
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2
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2
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2
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8
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