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Literature summary for 1.1.1.53 extracted from

  • Yang, J.K.; Park, M.S.; Waldo, G.S.; Suh, S.W.
    Directed evolution approach to a structural genomics project: Rv2002 from Mycobacterium tuberculosis (2003), Proc. Natl. Acad. Sci. USA, 100, 455-460.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
selenomethionine-substituted mutant enzyme I6T/V47M/T69K, expression in inclusion bodies in Escherichia coli B834(DE3) Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment) 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

Protein Variants

Protein Variants Comment 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 Comment Organism Structure
0.0033
-
progesterone pH 6.0, 30°C, reaction with NADH, mutant enzyme I6T/V47M/T69K Mycobacterium tuberculosis
0.017
-
20beta-hydroxyprogesterone pH 6.0, 30°C, reaction with NAD+, mutant enzyme I6T/V47M/T69K Mycobacterium tuberculosis
0.024
-
androsterone pH 6.0, 30°C, reaction with NAD+, mutant enzyme I6T/V47M/T69K Mycobacterium tuberculosis

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P9WGT1 Rv2002
-
Mycobacterium tuberculosis H37Rv P9WGT1 Rv2002
-

Purification (Commentary)

Purification (Comment) Organism
-
Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
17beta-estradiol + NAD+ 4% of the activity with androsterone Mycobacterium tuberculosis estrone + NADH + H+
-
?
17beta-estradiol + NAD+ 4% of the activity with androsterone Mycobacterium tuberculosis H37Rv estrone + NADH + H+
-
?
20alpha-hydroxyprogesterone + NAD+ 2% of the activity with androsterone Mycobacterium tuberculosis ? + NADH
-
?
20alpha-hydroxyprogesterone + NAD+ 2% of the activity with androsterone Mycobacterium tuberculosis H37Rv ? + NADH
-
?
20beta-hydroxyprogesterone + NAD+ 69% of the activity with androsterone Mycobacterium tuberculosis ? + NADH
-
?
20beta-hydroxyprogesterone + NAD+ 69% of the activity with androsterone Mycobacterium tuberculosis H37Rv ? + NADH
-
?
androsterone + NAD+
-
Mycobacterium tuberculosis ? + NADH
-
?
androsterone + NAD+
-
Mycobacterium tuberculosis H37Rv ? + NADH
-
?
epiandrosterone + NAD+ 3% of the activity with androsterone Mycobacterium tuberculosis ? + NADH
-
?
epiandrosterone + NAD+ 3% of the activity with androsterone Mycobacterium tuberculosis H37Rv ? + NADH
-
?
progesterone + NADH 22% of the activity with androsterone 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 Comment Organism Structure
72
-
progesterone pH 6.0, 30°C, reaction with NADH Mycobacterium tuberculosis
258
-
20beta-hydroxyprogesterone pH 6.0, 30°C, reaction with NAD+ Mycobacterium tuberculosis
456
-
androsterone pH 6.0, 30°C, reaction with NAD+ Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment 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 Comment Organism Structure
NADH preference of NADH over NADPH Mycobacterium tuberculosis