Cloned (Comment) | Organism |
---|---|
selenomethionine-substituted mutant enzyme I6T/V47M/T69K, expression in inclusion bodies in Escherichia coli B834(DE3) | Mycobacterium tuberculosis |
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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WGT1 | Rv2002 | - |
Mycobacterium tuberculosis H37Rv | P9WGT1 | Rv2002 | - |
Purification (Comment) | Organism |
---|---|
- |
Mycobacterium tuberculosis |
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 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 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 | Comment | Organism | Structure |
---|---|---|---|
NADH | preference of NADH over NADPH | Mycobacterium tuberculosis |