1.1.1.145: 3beta-hydroxy-DELTA5-steroid dehydrogenase

This is an abbreviated version, for detailed information about 3beta-hydroxy-DELTA5-steroid dehydrogenase, go to the full flat file.

Reaction

a 3beta-hydroxy-DELTA5-steroid
+
NAD+
=
a 3-oxo-DELTA5-steroid
+
NADH
+
H+

Synonyms

3-beta-hydroxy-5-ene steroid dehydrogenase, 3-beta-hydroxy-delta(5)-steroid dehydrogenase, 3beta hydroxysteroid dehydrogenase, 3beta-HSD, 3beta-HSD type II, 3beta-HSD1, 3beta-HSD2, 3beta-HSD22, 3beta-HSDH, 3beta-HSD_1, 3beta-hydroxy steroid dehydrogenase/5-ene-4-ene isomerase, 3beta-hydroxy steroid dehydrogenase/isomerase, 3beta-hydroxy-5-ene steroid dehydrogenase, 3beta-hydroxy-5-ene-steroid dehydrogenase, 3beta-hydroxy-5-ene-steroid oxidoreductase, 3beta-hydroxy-DELTA5-C27-steroid dehydrogenase/isomerase, 3beta-hydroxy-DELTA5-C27-steroid oxidoreductase, 3beta-hydroxysteroid dehydrogenase, 3beta-hydroxysteroid dehydrogenase 1, 3beta-hydroxysteroid dehydrogenase type 1, 3beta-hydroxysteroid dehydrogenase type 2, 3beta-hydroxysteroid dehydrogenase type II, 3beta-hydroxysteroid dehydrogenase type1, 3beta-hydroxysteroid dehydrogenase/DELTA5-DELAT4 isomerase type 2, 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase, 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4-isomerase, 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4-isomerase type II, 3beta-hydroxysteroid dehydrogenase/isomerase, 3beta-hydroxysteroid dehydrogenase/isomerase type 1, 3betaHSD, 3betaHSD1, 3betaHSD2, 5-ene-3-beta-hydroxysteroid dehydrogenase, 5alpha-dihydrotestosterone 3beta-hydroxysteroid dehydrogenase, dehydrogenase, 3beta-hydroxy-DELTA5-steroid, DELTA5-3beta-hydroxysteroid dehydrogenase, DHRS4, HSD3B, HSD3B2, HSDB1, More, progesterone reductase, steroid-DELTA5-3beta-ol dehydrogenase, type 1 3beta-hydroxysteroid dehydrogenase, type 1 3beta-hydroxysteroid dehydrogenase/isomerase, type 2 3beta-hydroxysteroid dehydrogenase, type 2 3beta-hydroxysteroid dehydrogenase/isomerase

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.1 With NAD+ or NADP+ as acceptor
                1.1.1.145 3beta-hydroxy-DELTA5-steroid dehydrogenase

Engineering

Engineering on EC 1.1.1.145 - 3beta-hydroxy-DELTA5-steroid dehydrogenase

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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C111A
-
contrary to wild-type, presence of 2-mercaptoethanol does not decrease Km value for substrate and NAD+
C111S
-
contrary to wild-type, presence of 2-mercaptoethanol does not decrease Km value for substrate and NAD+
C72F
-
contrary to wild-type, presence of 2-mercaptoethanol does not decrease Km value for substrate and NAD+
C72S
-
contrary to wild-type, presence of 2-mercaptoethanol does not decrease Km value for substrate and NAD+
C83A
-
2-5fold increase in Km values
C83S
-
no catalytic acitivity; similar to wild-type, presence of 2-mercaptoethanol results in 2.5fold decrease of Km values
D241N
-
39% of wild-type activity
D257L
-
trace activity
D258L
-
trace activity
D265N
-
12% of wild-type activity
D35A/K36R
-
the D35A/K36R mutant enzyme of 3beta-HSD1 has no measurable 3beta-HSD activity with NADH as the cofactor and now uses NADPH as the cofactor and NADH specificity of the wild-type 1 isomerase is completely lost
D36A/K37R
-
uses NADP+ as cofactor, no activity with NAD+
D61N
-
D61N mutation produces a 1.3fold increase in Km for the dehydroepiandrosterone substrate, a 1.9fold increase in Km for NAD+ utilization by the 3beta-HSD activity and little change in isomerase kinetics for either substrate or cofactor
D61V
-
mutant shows no enzymatic activity
DELTA283-310
-
cytosolic enzyme
E126L
site-directed mutagenesis, functional characterization
E192A
-
the E192A mutation causes an 8.2fold increase in Km (dehydroepiandrosterone) and no change in the Km for NAD+
H154F
no activity
H232A
site-directed mutagenesis, functional characterization
L341P
-
mutation in HSD3B2 gene identified in neonate with classical 3beta-hydroxysteroid dehydrogenase type II deficiency and under-virilization. About 6% of wild-type activity, putative structural alteration in protein
M187T
-
the 3beta-HSD1 mutant shows 16fold lower affinity inhibition by trilostane compared to the wild type enzyme, and produces a 2.2fold decrease in the Km value for the NAD+ utilization by of 3beta-HSD1 and a 9.2fold decrease in the Km measured for NADH
N100A
site-directed mutagenesis, functional characterization
N100S
site-directed mutagenesis, functional characterization
N323L
site-directed mutagenesis, functional characterization
Q105M
site-directed mutagenesis of isozyme 1, the mutants show altered kinetics compared to the wild-type isozymes; site-directed mutagenesis of isozyme 2, the mutants show altered kinetics compared to the wild-type isozymes
Q105M1
5fold decrease in catalytic efficiency; mutant retains the lower affinity profile of inhibition by epostane that is observed with wild-type 3beta-HSD2
Q240R
site-directed mutagenesis of isozyme 1, the mutant shows altered kinetics compared to the wild-type isozyme
R195R
-
the P195R mutation shifts the low-affinity, noncompetitive inhibition profile of 3beta-HSD2 to a high affinity (Ki= 0.19 microM), competitive inhibition profile. The P195R mutant has a 12.2fold lower Km value for 16beta-hydroxy-dehydroepiandrosterone compared to wild-type 3beta-HSD2. The coenzyme values measured for P195R mutant are similar to those measured for wildtype 3beta-HSD2
R240Q
site-directed mutagenesis of isozyme 2, the mutant shows altered kinetics compared to the wild-type isozyme
R335stop
-
mutation in HSD3B2 gene identified in neonate with classical 3beta-hydroxysteroid dehydrogenase type II deficiency and under-virilization. No conversion of pregnenolone or dehydroepiandrosterone
S123A
-
24% of wild-type 3beta-HSD1 activity
S124A
-
no detectable 3beta-HSD1 activity
S124T
-
mutant shows activity similar to the wild type enzyme
S322A
site-directed mutagenesis, functional characterization
T177N
-
significantly improved stability at cold temperature, presence of NAD+ is required for complete stability at 0C
T8A
-
mutant shows almost similar kinetic values as wild-type
W355stop
-
mutation in HSD3B2 gene identified in neonate with classical 3beta-hydroxysteroid dehydrogenase type II deficiency and under-virilization. No conversion of pregnenolone or dehydroepiandrosterone
additional information