BRENDA - Enzyme Database show
show all sequences of 1.1.1.53

Directed evolution approach to a structural genomics project: Rv2002 from Mycobacterium tuberculosis

Yang, J.K.; Park, M.S.; Waldo, G.S.; Suh, S.W.; Proc. Natl. Acad. Sci. USA 100, 455-460 (2003)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
selenomethionine-substituted mutant enzyme I6T/V47M/T69K, expression in inclusion bodies in Escherichia coli B834(DE3)
Mycobacterium tuberculosis
Crystallization (Commentary)
Crystallization
Organism
crystal structure of the mutant enzyme I6T/V47M/T69K in binary complex with NAD+ at 1.8 A resolution and as a ternary complex with androsterone and NADH at 2.4 A
Mycobacterium tuberculosis
Engineering
Amino acid exchange
Commentary
Organism
I6T/V47M/T69K
mutant enzyme with a maximal improvement in solubility, high catalytic activity as a NADP-dependent 3alpha,20beta-hydroxysteroid dehydrogenase
Mycobacterium tuberculosis
I6T/V47M/T69K/E142A
pH optimum for reduction of progesterone shifts slightly from pH 6.0 to pH 6.5 compared to for mutant enzyme I6T/V47M/T69K, the oxidation of androsterone shows a dual pH optimum, one at pH 6.0-6.5 and the other at pH 10
Mycobacterium tuberculosis
I6T/V47M/T69K/S140A
both oxidation of androsterone and reduction of progesterone are completely lost
Mycobacterium tuberculosis
I6T/V47M/T69K/Y153F
both oxidation of androsterone and reduction of progesterone are completely lost
Mycobacterium tuberculosis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0033
-
progesterone
pH 6.0, 30C, reaction with NADH, mutant enzyme I6T/V47M/T69K
Mycobacterium tuberculosis
0.017
-
20beta-hydroxyprogesterone
pH 6.0, 30C, reaction with NAD+, mutant enzyme I6T/V47M/T69K
Mycobacterium tuberculosis
0.024
-
androsterone
pH 6.0, 30C, reaction with NAD+, mutant enzyme I6T/V47M/T69K
Mycobacterium tuberculosis
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mycobacterium tuberculosis
P9WGT1
Rv2002
-
Mycobacterium tuberculosis H37Rv
P9WGT1
Rv2002
-
Purification (Commentary)
Commentary
Organism
-
Mycobacterium tuberculosis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
17beta-estradiol + NAD+
4% of the activity with androsterone
657157
Mycobacterium tuberculosis
estrone + NADH
-
-
-
?
20alpha-hydroxyprogesterone + NAD+
2% of the activity with androsterone
657157
Mycobacterium tuberculosis
? + NADH
-
-
-
?
20beta-hydroxyprogesterone + NAD+
69% of the activity with androsterone
657157
Mycobacterium tuberculosis
? + NADH
-
-
-
?
androsterone + NAD+
-
657157
Mycobacterium tuberculosis
? + NADH
-
-
-
?
epiandrosterone + NAD+
3% of the activity with androsterone
657157
Mycobacterium tuberculosis
? + NADH
-
-
-
?
progesterone + NADH
22% of the activity with androsterone
657157
Mycobacterium tuberculosis
20beta-hydroxy-pregn-4-ene-3-one + NAD+
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
72
-
progesterone
pH 6.0, 30C, reaction with NADH
Mycobacterium tuberculosis
258
-
20beta-hydroxyprogesterone
pH 6.0, 30C, reaction with NAD+
Mycobacterium tuberculosis
456
-
androsterone
pH 6.0, 30C, reaction with NAD+
Mycobacterium tuberculosis
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6
6.5
oxidation of androsterone shows a dual pH-optimum, one at pH 6.0-6.5 and the other at pH 10, mutant enzyme I6T/V47M/T69K/E142A
Mycobacterium tuberculosis
6
-
progesterone + NADH, mutant enzyme I6T/V47M/T69K
Mycobacterium tuberculosis
6.5
-
progesterone + NADH, mutant enzyme I6T/V47M/T69K/E142A
Mycobacterium tuberculosis
10
-
oxidation of androsterone shows a dual pH-optimum, one at pH 6.0-6.5 and the other at pH 10, mutant enzyme I6T/V47M/T69K/E142A
Mycobacterium tuberculosis
Cofactor
Cofactor
Commentary
Organism
Structure
NADH
preference of NADH over NADPH
Mycobacterium tuberculosis
Cloned(Commentary) (protein specific)
Commentary
Organism
selenomethionine-substituted mutant enzyme I6T/V47M/T69K, expression in inclusion bodies in Escherichia coli B834(DE3)
Mycobacterium tuberculosis
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADH
preference of NADH over NADPH
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
crystal structure of the mutant enzyme I6T/V47M/T69K in binary complex with NAD+ at 1.8 A resolution and as a ternary complex with androsterone and NADH at 2.4 A
Mycobacterium tuberculosis
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
I6T/V47M/T69K
mutant enzyme with a maximal improvement in solubility, high catalytic activity as a NADP-dependent 3alpha,20beta-hydroxysteroid dehydrogenase
Mycobacterium tuberculosis
I6T/V47M/T69K/E142A
pH optimum for reduction of progesterone shifts slightly from pH 6.0 to pH 6.5 compared to for mutant enzyme I6T/V47M/T69K, the oxidation of androsterone shows a dual pH optimum, one at pH 6.0-6.5 and the other at pH 10
Mycobacterium tuberculosis
I6T/V47M/T69K/S140A
both oxidation of androsterone and reduction of progesterone are completely lost
Mycobacterium tuberculosis
I6T/V47M/T69K/Y153F
both oxidation of androsterone and reduction of progesterone are completely lost
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0033
-
progesterone
pH 6.0, 30C, reaction with NADH, mutant enzyme I6T/V47M/T69K
Mycobacterium tuberculosis
0.017
-
20beta-hydroxyprogesterone
pH 6.0, 30C, reaction with NAD+, mutant enzyme I6T/V47M/T69K
Mycobacterium tuberculosis
0.024
-
androsterone
pH 6.0, 30C, reaction with NAD+, mutant enzyme I6T/V47M/T69K
Mycobacterium tuberculosis
Purification (Commentary) (protein specific)
Commentary
Organism
-
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
17beta-estradiol + NAD+
4% of the activity with androsterone
657157
Mycobacterium tuberculosis
estrone + NADH
-
-
-
?
20alpha-hydroxyprogesterone + NAD+
2% of the activity with androsterone
657157
Mycobacterium tuberculosis
? + NADH
-
-
-
?
20beta-hydroxyprogesterone + NAD+
69% of the activity with androsterone
657157
Mycobacterium tuberculosis
? + NADH
-
-
-
?
androsterone + NAD+
-
657157
Mycobacterium tuberculosis
? + NADH
-
-
-
?
epiandrosterone + NAD+
3% of the activity with androsterone
657157
Mycobacterium tuberculosis
? + NADH
-
-
-
?
progesterone + NADH
22% of the activity with androsterone
657157
Mycobacterium tuberculosis
20beta-hydroxy-pregn-4-ene-3-one + NAD+
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
72
-
progesterone
pH 6.0, 30C, reaction with NADH
Mycobacterium tuberculosis
258
-
20beta-hydroxyprogesterone
pH 6.0, 30C, reaction with NAD+
Mycobacterium tuberculosis
456
-
androsterone
pH 6.0, 30C, reaction with NAD+
Mycobacterium tuberculosis
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6
6.5
oxidation of androsterone shows a dual pH-optimum, one at pH 6.0-6.5 and the other at pH 10, mutant enzyme I6T/V47M/T69K/E142A
Mycobacterium tuberculosis
6
-
progesterone + NADH, mutant enzyme I6T/V47M/T69K
Mycobacterium tuberculosis
6.5
-
progesterone + NADH, mutant enzyme I6T/V47M/T69K/E142A
Mycobacterium tuberculosis
10
-
oxidation of androsterone shows a dual pH-optimum, one at pH 6.0-6.5 and the other at pH 10, mutant enzyme I6T/V47M/T69K/E142A
Mycobacterium tuberculosis
Other publictions for EC 1.1.1.53
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)
738928
Zhang
Human 3-alpha hydroxysteroid d ...
Homo sapiens
J. Steroid Biochem. Mol. Biol.
141
135-143
2014
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1
1
1
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4
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2
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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
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4
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4
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4
4
726321
Tokarz
Zebrafish 20beta-hydroxysteroi ...
Danio rerio
PLoS ONE
8
e54851
2013
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1
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1
1
1
1
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-
725963
Tokarz
Discovery of a novel enzyme me ...
Danio rerio
Mol. Cell. Endocrinol.
349
202-213
2012
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-
-
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1
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
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-
698084
Sreenivasulu
New evidences for the involvem ...
Clarias gariepinus
Gen. Comp. Endocrinol.
163
259-269
2009
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1
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3
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1
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6
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1
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10
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1
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10
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1
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686368
Albertsson
Proteomic analyses indicate in ...
Oncorhynchus mykiss
Ecotoxicol. Environ. Safety
68
33-39
2007
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-
-
-
-
-
-
-
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1
-
-
2
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2
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1
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2
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1
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1
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-
687870
Hoffmann
Understanding oligomerization ...
Comamonas testosteroni
J. Biotechnol.
129
131-139
2007
-
-
1
-
-
-
-
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-
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1
1
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3
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1
1
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1
2
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1
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1
1
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1
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1
2
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-
688455
Mittelholzer
20 beta-hydroxysteroid dehydro ...
Gadus morhua
J. Mol. Endocrinol.
39
319-328
2007
-
-
1
-
-
-
-
-
-
-
-
1
-
1
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7
-
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1
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1
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1
1
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1
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7
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1
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668684
Sreenivasulu
Expression of 20beta-hydroxyst ...
Channa striata, Gallus gallus
Fish Physiol. Biochem.
31
227-230
2006
1
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6
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12
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3
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2
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1
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2
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12
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3
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-
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-
668714
Nakamura
Changes in steroidogenic enzym ...
Oncorhynchus mykiss
Gen. Comp. Endocrinol.
144
224-231
2005
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3
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1
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1
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1
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1
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-
657157
Yang
Directed evolution approach to ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Proc. Natl. Acad. Sci. USA
100
455-460
2003
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1
1
4
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3
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5
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1
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6
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4
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1
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1
1
1
4
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1
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6
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3
4
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654932
Senthilkumaran
Ovarian carbonyl reductase-lik ...
Oreochromis niloticus
Biol. Reprod.
67
1080-1086
2002
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1
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1
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5
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4
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7
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655669
Wang
Cloning of zebrafish ovarian c ...
Danio rerio
Gen. Comp. Endocrinol.
127
209-216
2002
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1
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5
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2
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287111
Kazeto
20beta-hydroxysteroid dehydrog ...
Anguilla japonica
Gen. Comp. Endocrinol.
122
109-115
2001
1
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2
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2
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1
1
4
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1
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287102
Takada
Further studies on 20beta-hydr ...
Rattus norvegicus
Life Sci.
67
2075-2082
2000
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1
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2
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2
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1
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5
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2
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1
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5
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287107
Guan
Isoleucine-15 of rainbow trout ...
Oncorhynchus mykiss
Proc. Natl. Acad. Sci. USA
97
3079-3083
2000
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1
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3
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1
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3
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1
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2
3
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3
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1
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2
3
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2
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287101
Takada
20beta-Hydroxysteroid dehydrog ...
Rattus norvegicus
J. Pharmacol. Exp. Ther.
287
504-507
1998
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1
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2
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2
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1
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2
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6
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1
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1
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6
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287108
Vizziano
20beta-Hydroxysteroid dehydrog ...
Oncorhynchus mykiss
Biol. Reprod.
54
1-7
1996
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287109
Ghosh
Mechanism of inhibition of 3al ...
Streptomyces exfoliatus
Curr. Biol.
2
973-980
1994
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287110
Ghosh
The refined three-dimensional ...
Streptomyces exfoliatus
Structure
2
629-640
1994
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287100
Krook
Short-chain dehydrogenases. Pr ...
Streptomyces exfoliatus
Eur. J. Biochem.
209
233-239
1992
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287103
Ohno
Ontogeny of testicular steroid ...
Sus scrofa
J. Steroid Biochem. Mol. Biol.
42
17-21
1992
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287106
Tanaka
Pig testicular 20beta-hydroxys ...
Sus scrofa
J. Biol. Chem.
267
13451-13455
1992
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287097
Ohno
20beta-Hydroxysteroid dehydrog ...
Sus scrofa
J. Steroid Biochem. Mol. Biol.
38
787-794
1991
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287098
Ohno
20beta-Hydroxysteroid dehydrog ...
Sus scrofa
Chem. Pharm. Bull.
39
972-975
1991
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287083
Tyrakowska
Enzyme kinetics in reversed mi ...
Streptomyces exfoliatus
Eur. J. Biochem.
187
81-88
1990
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287104
Adachi
-
Involvement of pH in the enhan ...
Oncorhynchus masou rhodurus, Oncorhynchus mykiss
Biomed. Res.
11
327-335
1990
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287084
Nakajin
20beta-Hydroxysteroid dehydrog ...
Sus scrofa
J. Biochem.
104
565-569
1988
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287085
Carrea
Properties of immobilized subu ...
Streptomyces exfoliatus
Methods Enzymol.
135
475-483
1987
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287086
Vecchio
Multiparameter study of denatu ...
Streptomyces exfoliatus
Biochim. Biophys. Acta
914
122-126
1987
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287087
Carrea
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Enzyme-catalyzed steroid trans ...
Streptomyces exfoliatus
Methods Enzymol.
136
150-157
1987
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287088
Carrea
Effect of the lyotropic series ...
Streptomyces exfoliatus
Biochim. Biophys. Acta
784
16-23
1984
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287089
Strickler
Study of 3alpha, 20beta-hydrox ...
Streptomyces exfoliatus
Biochemistry
19
4950-4954
1980
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287090
Sweet
Bifunctional enzyme activity a ...
Streptomyces exfoliatus
Biochemistry
19
978-986
1980
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287091
Monder
Reduction of steroidal 20-oxo- ...
Streptomyces exfoliatus
J. Steroid Biochem.
9
1229-1231
1978
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287092
Sweet
Substrate characteristics of p ...
Streptomyces exfoliatus
Biochem. Biophys. Res. Commun.
83
187-194
1978
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287093
Edwards
Comparison of the 3alpha-and 2 ...
Streptomyces exfoliatus
Biochemistry
17
4370-4376
1978
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287094
Szymanski
20beta-Hydroxysteroid oxidored ...
Streptomyces exfoliatus
J. Biol. Chem.
252
205-211
1977
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287099
Blomquist
-
The molecular weight and subst ...
Streptomyces exfoliatus
Arch. Biochem. Biophys.
159
590-595
1973
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287095
Hubner
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20beta-hydroxy-steroid-dehydro ...
Streptomyces exfoliatus
Biochem. Z.
333
95-105
1960
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287096
Lynn
The conversion of progesterone ...
Cavia porcellus
J. Biol. Chem.
232
1015-1030
1958
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