BRENDA - Enzyme Database show
show all sequences of 1.3.99.4

Purification, crystallization and preliminary X-ray crystallographic analysis of 3-ketosteroid DELTA1-dehydrogenase from Rhodococcus erythropolis SQ1

Rohman, A.; van Oosterwijk, N.; Dijkstra, B.W.; Acta Crystallogr. Sect. F 68, 551-556 (2012)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene kstD1, heterologous expression of the His-tagged DELTA1-KSTD1 protein in Escherichia coli strain BL21(DE3)
Rhodococcus erythropolis
Crystallization (Commentary)
Crystallization
Organism
sitting drop vapour diffusion method, mixing of 15 mg/ml protein in 25 mM bicine, pH 9.0, 100 mM NaCl, 10% v/v glycerol with 2% v/v PEG 400, 0.1 M, HEPES, pH 7.5, 2.0 M ammonium sulfate, 20°C, 5-7 days, method optimization, X-ray diffraction structure determination and analysis at 2.0 A resolution, multiwavelength anomalous dispersion analysis
Rhodococcus erythropolis
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
56000
-
x * 56000
Rhodococcus erythropolis
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4-androstene-3,17-dione + FAD
Rhodococcus erythropolis
-
1,4-androstadiene-3,17-dione + FADH2
-
-
?
4-androstene-3,17-dione + FAD
Rhodococcus erythropolis SQ1
-
1,4-androstadiene-3,17-dione + FADH2
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Rhodococcus erythropolis
Q9RA02
three 3-ketosteroid DELTA1-dehydrogenase isoenzymes, gene kstD1 encodes isozyme DELTA1-KSTD1
-
Rhodococcus erythropolis SQ1
Q9RA02
three 3-ketosteroid DELTA1-dehydrogenase isoenzymes, gene kstD1 encodes isozyme DELTA1-KSTD1
-
Purification (Commentary)
Commentary
Organism
recombinant His-tagged DELTA1-KSTD1 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
Rhodococcus erythropolis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-androstene-3,17-dione + FAD
-
723868
Rhodococcus erythropolis
1,4-androstadiene-3,17-dione + FADH2
-
-
-
?
4-androstene-3,17-dione + FAD
-
723868
Rhodococcus erythropolis SQ1
1,4-androstadiene-3,17-dione + FADH2
-
-
-
?
Subunits
Subunits
Commentary
Organism
?
x * 56000
Rhodococcus erythropolis
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
6
9
below pH 6.0 no apparent Tm is observed, indicating that DELTA1-KSTD1 is destabilized at low pH. The higher the pH value, within the range of pH 6.0-9.0, the higher the apparent Tm of DELTA1-KSTD1. At pH 9.0 the apparent Tm of DELTA1-KSTD1 is about 7 K higher than at pH 7.0
Rhodococcus erythropolis
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
prosthetic group, flavoenzyme
Rhodococcus erythropolis
Cloned(Commentary) (protein specific)
Commentary
Organism
gene kstD1, heterologous expression of the His-tagged DELTA1-KSTD1 protein in Escherichia coli strain BL21(DE3)
Rhodococcus erythropolis
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
prosthetic group, flavoenzyme
Rhodococcus erythropolis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
sitting drop vapour diffusion method, mixing of 15 mg/ml protein in 25 mM bicine, pH 9.0, 100 mM NaCl, 10% v/v glycerol with 2% v/v PEG 400, 0.1 M, HEPES, pH 7.5, 2.0 M ammonium sulfate, 20°C, 5-7 days, method optimization, X-ray diffraction structure determination and analysis at 2.0 A resolution, multiwavelength anomalous dispersion analysis
Rhodococcus erythropolis
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
56000
-
x * 56000
Rhodococcus erythropolis
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4-androstene-3,17-dione + FAD
Rhodococcus erythropolis
-
1,4-androstadiene-3,17-dione + FADH2
-
-
?
4-androstene-3,17-dione + FAD
Rhodococcus erythropolis SQ1
-
1,4-androstadiene-3,17-dione + FADH2
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His-tagged DELTA1-KSTD1 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
Rhodococcus erythropolis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-androstene-3,17-dione + FAD
-
723868
Rhodococcus erythropolis
1,4-androstadiene-3,17-dione + FADH2
-
-
-
?
4-androstene-3,17-dione + FAD
-
723868
Rhodococcus erythropolis SQ1
1,4-androstadiene-3,17-dione + FADH2
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 56000
Rhodococcus erythropolis
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
6
9
below pH 6.0 no apparent Tm is observed, indicating that DELTA1-KSTD1 is destabilized at low pH. The higher the pH value, within the range of pH 6.0-9.0, the higher the apparent Tm of DELTA1-KSTD1. At pH 9.0 the apparent Tm of DELTA1-KSTD1 is about 7 K higher than at pH 7.0
Rhodococcus erythropolis
General Information
General Information
Commentary
Organism
physiological function
3-ketosteroid DELTA1-dehydrogenase plays a crucial role in the early steps of steroid degradation by introducing a double bond between the C1 and C2 atoms of the A-ring of its 3-ketosteroid substrates
Rhodococcus erythropolis
General Information (protein specific)
General Information
Commentary
Organism
physiological function
3-ketosteroid DELTA1-dehydrogenase plays a crucial role in the early steps of steroid degradation by introducing a double bond between the C1 and C2 atoms of the A-ring of its 3-ketosteroid substrates
Rhodococcus erythropolis
Other publictions for EC 1.3.99.4
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)
743236
Guevara
Functional differentiation of ...
Rhodococcus ruber, Rhodococcus ruber Chol-4
Microb. Cell Fact.
16
42
2017
-
-
-
-
-
-
-
20
-
-
-
-
-
7
-
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33
1
-
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20
-
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-
33
3
-
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-
-
-
-
-
-
-
-
-
-
-
743031
Shao
A mutant form of 3-ketosteroi ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum JC-12, Mycolicibacterium neoaurum ST-095
J. Ind. Microbiol. Biotechnol.
43
691-701
2016
-
-
1
-
1
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2
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4
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1
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3
1
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1
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1
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1
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3
1
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2
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-
2
2
743221
Yao
Characterization and engineer ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum ATCC 25795
Metab. Eng.
24
181-191
2014
-
1
1
-
-
-
-
14
-
-
-
-
-
6
-
-
-
-
-
-
-
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28
1
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3
3
-
-
-
-
-
-
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14
-
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-
-
-
-
-
-
-
-
28
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
724670
Brzezinska
The role of 3-ketosteroid 1(2) ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
BMC Microbiol.
13
43
2013
-
-
-
-
-
-
-
-
-
-
-
-
-
163
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
1
1
-
-
725830
Zhang
Bioconversion of 4-androstene- ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum JC-12
J. Steroid Biochem. Mol. Biol.
135
36-42
2013
-
-
1
-
-
-
-
-
-
-
-
2
-
3
-
-
-
-
-
-
1
-
4
-
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
2
-
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-
-
1
-
4
-
1
-
-
-
1
-
-
-
-
1
1
-
-
-
742850
Rohman
Crystal structure and site-di ...
Rhodococcus erythropolis, Rhodococcus erythropolis SQ1
J. Biol. Chem.
288
35559-35568
2013
-
-
1
1
3
-
-
3
-
-
-
-
-
4
-
-
-
-
-
-
1
-
6
-
-
-
-
3
-
-
-
1
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-
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-
-
1
1
1
3
-
-
-
-
3
-
-
-
-
-
-
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-
-
1
-
6
-
-
-
-
3
-
-
-
-
-
-
-
-
3
3
723868
Rohman
Purification, crystallization ...
Rhodococcus erythropolis, Rhodococcus erythropolis SQ1
Acta Crystallogr. Sect. F
68
551-556
2012
-
-
1
1
-
-
-
-
-
-
1
2
-
4
-
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1
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2
1
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1
1
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1
1
1
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1
2
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1
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-
-
-
2
1
-
-
-
-
-
-
1
-
-
1
1
-
-
-
724074
Chen
Characterization and applicati ...
Aspergillus fumigatus, Aspergillus fumigatus CICC 40167
Appl. Microbiol. Biotechnol.
96
133-142
2012
-
1
1
-
-
-
-
-
1
-
1
2
-
2
-
-
1
-
-
-
-
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2
1
1
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1
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2
-
1
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1
1
2
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1
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1
2
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-
1
-
-
-
-
2
1
1
-
-
-
1
-
-
1
-
1
1
-
-
-
724000
Wei
Inactivation and augmentation ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum NwIB-01
Appl. Environ. Microbiol.
76
4578-4582
2010
-
1
1
-
-
-
-
-
-
-
-
2
-
2
-
-
1
-
-
-
1
-
6
-
1
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1
-
-
1
-
-
-
-
1
1
1
-
-
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2
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1
-
-
1
-
6
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
684994
Knol
3-Keto-5alpha-steroid DELTA(1) ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv, Rhodococcus erythropolis, Rhodococcus erythropolis SQ1
Biochem. J.
410
339-346
2008
-
-
1
-
-
1
-
23
-
-
3
-
-
172
-
-
1
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-
-
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49
-
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3
-
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1
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23
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3
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1
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-
-
49
-
-
-
-
-
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-
-
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-
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-
-
-
688748
Li
Expression of ksdD gene encodi ...
Pimelobacter simplex
Lett. Appl. Microbiol.
44
563-568
2007
-
-
1
-
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-
-
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-
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1
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4
-
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2
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1
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1
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2
-
1
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-
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-
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-
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-
675829
Brzostek
Identification and targeted di ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Ra / ATCC 25177, Mycolicibacterium smegmatis, Mycolicibacterium smegmatis mc(2)155 / ATCC 700084
Microbiology
151
2393-2402
2005
-
2
2
-
1
-
-
-
-
-
-
-
-
31
-
-
-
-
-
-
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2
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1
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2
2
1
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1
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-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
654270
Horinouchi
A new bacterial steroid degrad ...
Comamonas testosteroni, Comamonas testosteroni TA441
Appl. Environ. Microbiol.
69
4421-4430
2003
-
-
-
-
-
-
-
-
-
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2
-
2
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2
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657152
Adham
-
Biochemical studies on the mic ...
Pseudomonas fluorescens
Proc. Biochem.
38
897-902
2003
1
-
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5
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6
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1
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1
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1
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656722
van der Geize
Molecular and functional chara ...
Rhodococcus erythropolis, Rhodococcus erythropolis SQ1
Microbiology
148
3285-3292
2002
-
-
1
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2
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2
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6
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8
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1
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391391
Van der Geize
Unmarked gene deletion mutagen ...
Rhodococcus erythropolis
FEMS Microbiol. Lett.
205
197-202
2001
-
-
1
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-
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6
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1
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391394
Fujii
Essential tyrosine residues in ...
Rhodococcus rhodochrous
J. Biochem.
126
662-667
1999
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1
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1
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1
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1
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391396
Morii
3-Ketosteroid-DELTA1-dehydroge ...
Rhodococcus rhodochrous
J. Biochem.
124
1026-1032
1998
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1
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1
1
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1
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5
-
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1
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1
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1
1
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1
1
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1
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1
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391392
Choi
Purification and characterizat ...
Pimelobacter simplex
J. Biochem.
117
1043-1049
1995
-
-
-
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5
1
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1
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1
1
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1
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1
5
1
1
1
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1
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5
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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
5
1
1
1
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1
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391393
Choi
Secretory overproduction of Ar ...
Pimelobacter simplex
Appl. Microbiol. Biotechnol.
43
1044-1049
1995
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391395
Molnar
Molecular cloning, expression ...
Pimelobacter simplex
Mol. Microbiol.
15
895-905
1995
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1
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391375
Itagaki
Purification and characterizat ...
Gordonia rubripertincta
Biochim. Biophys. Acta
1038
60-67
1990
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2
11
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15
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11
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1
15
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2
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391378
Itagaki
Spectral properties of 3-ketos ...
Gordonia rubripertincta
Biochim. Biophys. Acta
1040
281-286
1990
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391379
Kloosterman IV
-
Effect of supersaturated aqueo ...
Pimelobacter simplex
Enzyme Microb. Technol.
6
113-116
1984
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391385
Udvardy
-
Kinetics of the formation of s ...
Pimelobacter simplex
FEMS Symp.
13
21-33
1982
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391384
Atrat
Steroid transformation with im ...
Mycolicibacterium phlei, Nocardia erythropolis, Rhodococcus opacus
Z. Allg. Mikrobiol.
21
3-6
1981
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3
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3
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3
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391376
Atrat
-
Efficient purification of a mi ...
Rhodococcus opacus
J. Chromatogr.
189
279-283
1980
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391389
Hörhold
Steroid transforming enzymes i ...
Rhodococcus opacus
Z. Allg. Mikrobiol.
20
23-32
1980
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391377
Groh
Steroid-transforming enzymes f ...
Rhodococcus opacus
Z. Allg. Mikrobiol.
19
727-730
1979
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3
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2
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391388
Hörhold
Steroid-transforming enzymes f ...
Rhodococcus opacus
Z. Allg. Mikrobiol.
19
731-739
1979
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391387
Lestrovaja
Steroid transforming enzymes f ...
Rhodococcus opacus
Z. Allg. Mikrobiol.
18
189-196
1978
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1
1
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391390
Abul-Hajj
Isolation of vitamin K2(35) fr ...
Ilyonectria destructans, Pimelobacter simplex, Plectosphaerella cucumerina, Rhodococcus hoagii
J. Biol. Chem.
253
2356-2360
1978
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4
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15
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391386
Hörhold
Steroid-transforming enzymes f ...
Rhodococcus opacus
Z. Allg. Mikrobiol.
16
559-561
1976
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391380
Penasse
The active center of the 3-oxo ...
Pimelobacter simplex
Eur. J. Biochem.
47
555-559
1974
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391382
Rodig
The stereochemistry of hydroge ...
Saccharomyces cerevisiae
Biochem. Biophys. Res. Commun.
56
467-470
1974
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391381
Aries
-
Degradation of steroids by int ...
Clostridium paraputrificum
Biochim. Biophys. Acta
248
482-488
1971
-
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1
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5
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5
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1
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391383
Levy
Bacterial oxidation of steroid ...
Comamonas testosteroni
J. Biol. Chem.
234
2014-2021
1959
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8
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1
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1
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1
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9
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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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9
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1
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1
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