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Literature summary 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)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.53 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

EC Number Protein Variants Comment Organism
1.1.1.53 I6T/V47M/T69K mutant enzyme with a maximal improvement in solubility, high catalytic activity as a NADP-dependent 3alpha,20beta-hydroxysteroid dehydrogenase Mycobacterium tuberculosis
1.1.1.53 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
1.1.1.53 I6T/V47M/T69K/S140A both oxidation of androsterone and reduction of progesterone are completely lost Mycobacterium tuberculosis
1.1.1.53 I6T/V47M/T69K/Y153F both oxidation of androsterone and reduction of progesterone are completely lost Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.53 0.0033
-
 progesterone pH 6.0, 30°C, reaction with NADH, mutant enzyme I6T/V47M/T69K Mycobacterium tuberculosis
1.1.1.53 0.017
-
 20beta-hydroxyprogesterone pH 6.0, 30°C, reaction with NAD+, mutant enzyme I6T/V47M/T69K Mycobacterium tuberculosis
1.1.1.53 0.024
-
 androsterone pH 6.0, 30°C, reaction with NAD+, mutant enzyme I6T/V47M/T69K Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.53 Mycobacterium tuberculosis P9WGT1 Rv2002
-
1.1.1.53 Mycobacterium tuberculosis H37Rv P9WGT1 Rv2002
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.53
-
 Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.53 17beta-estradiol + NAD+ 4% of the activity with androsterone Mycobacterium tuberculosis estrone + NADH + H+
-
 ?
1.1.1.53 17beta-estradiol + NAD+ 4% of the activity with androsterone Mycobacterium tuberculosis H37Rv estrone + NADH + H+
-
 ?
1.1.1.53 20alpha-hydroxyprogesterone + NAD+ 2% of the activity with androsterone Mycobacterium tuberculosis ? + NADH
-
 ?
1.1.1.53 20alpha-hydroxyprogesterone + NAD+ 2% of the activity with androsterone Mycobacterium tuberculosis H37Rv ? + NADH
-
 ?
1.1.1.53 20beta-hydroxyprogesterone + NAD+ 69% of the activity with androsterone Mycobacterium tuberculosis ? + NADH
-
 ?
1.1.1.53 20beta-hydroxyprogesterone + NAD+ 69% of the activity with androsterone Mycobacterium tuberculosis H37Rv ? + NADH
-
 ?
1.1.1.53 androsterone + NAD+
-
 Mycobacterium tuberculosis ? + NADH
-
 ?
1.1.1.53 androsterone + NAD+
-
 Mycobacterium tuberculosis H37Rv ? + NADH
-
 ?
1.1.1.53 epiandrosterone + NAD+ 3% of the activity with androsterone Mycobacterium tuberculosis ? + NADH
-
 ?
1.1.1.53 epiandrosterone + NAD+ 3% of the activity with androsterone Mycobacterium tuberculosis H37Rv ? + NADH
-
 ?
1.1.1.53 progesterone + NADH 22% of the activity with androsterone Mycobacterium tuberculosis 20beta-hydroxy-pregn-4-ene-3-one + NAD+
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.53 72
-
 progesterone pH 6.0, 30°C, reaction with NADH Mycobacterium tuberculosis
1.1.1.53 258
-
 20beta-hydroxyprogesterone pH 6.0, 30°C, reaction with NAD+ Mycobacterium tuberculosis
1.1.1.53 456
-
 androsterone pH 6.0, 30°C, reaction with NAD+ Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.53 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
1.1.1.53 6
-
 progesterone + NADH, mutant enzyme I6T/V47M/T69K Mycobacterium tuberculosis
1.1.1.53 6.5
-
 progesterone + NADH, mutant enzyme I6T/V47M/T69K/E142A Mycobacterium tuberculosis
1.1.1.53 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

EC Number Cofactor Comment Organism Structure
1.1.1.53 NADH preference of NADH over NADPH Mycobacterium tuberculosis