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Literature summary for 1.1.1.213 extracted from

  • Chang, Y.H.; Chuang, L.Y.; Hwang, C.C.
    Mechanism of proton transfer in the 3alpha-hydroxysteroid dehydrogenase/carbonyl reductase from Comamonas testosteroni (2007), J. Biol. Chem., 282, 34306-34314.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Comamonas testosteroni

Protein Variants

Protein Variants Comment Organism
K159A site-directed mutagenesis, the mutation changes the rate-limiting step to the hydride transfer, proton transfer is blocked in the mutant but can be rescued using exogenous proton acceptors, such as buffers, small primary amines, and azide, overview Comamonas testosteroni
K159M site-directed mutagenesis, the mutation changes the rate-limiting step to the hydride transfer, proton transfer is blocked in the mutant but can be rescued using exogenous proton acceptors, such as buffers, small primary amines, and azide, overview Comamonas testosteroni

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics of wild-type and mutant enzymes in presence or absence of CAPS and methylamine, overview Comamonas testosteroni
0.0051
-
androsterone pH 10.4, 25°C, recombinant wild-type enzyme Comamonas testosteroni
0.033
-
androsterone pH 10.4, 25°C, recombinant mutant K159A Comamonas testosteroni

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
androsterone + NAD+ Comamonas testosteroni
-
androstanedione + NADH
-
r

Organism

Organism UniProt Comment Textmining
Comamonas testosteroni
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes ifrom Escherichia coli strain BL21(DE3) Comamonas testosteroni

Reaction

Reaction Comment Organism Reaction ID
a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ The overall oxidoreductive reaction comprises the deprotonation of tyrosine, proton abstraction by the tyrosinate anion, and hydride transfer from the hydroxysteroid to NAD+, followed by the release of a proton from the hydroxy group of tyrosine to the solution., The proton transfer to the external base, from Tyr155 to Lys159 via 2'-OH of ribose, with a late transition state is the rate-limiting step, mechanism, overview Comamonas testosteroni

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Comamonas testosteroni

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
androsterone + NAD+
-
Comamonas testosteroni androstanedione + NADH
-
r
androsterone + NAD+ the intramolecular proton transfer is a rate-limiting step, with a concomitant releasing of protons to bulk solvent Comamonas testosteroni androstanedione + NADH
-
r

Synonyms

Synonyms Comment Organism
3alpha-HSD/CR
-
Comamonas testosteroni
3alpha-hydroxysteroid dehydrogenase/carbonyl reductase
-
Comamonas testosteroni
More cf. EC 1.1.1.50 Comamonas testosteroni

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Comamonas testosteroni

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10.4
-
assay at Comamonas testosteroni

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Comamonas testosteroni
NADH
-
Comamonas testosteroni