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Literature summary extracted from

  • Doden, H.; Pollet, R.; Mythen, S.; Wawrzak, Z.; Devendran, S.; Cann, I.; Koropatkin, N.; Ridlon, J.
    Structural and biochemical characterization of 20-hydroxysteroid dehydrogenase from Bifidobacterium adolescentis strain L2-32 (2019), J. Biol. Chem., 294, 12040-12053 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.1.1.149 food industry the enzyme can alter glucocorticoid metabolism in the gut and thereby serves as potential probiotics for the management of androgen-dependent diseases Bifidobacterium adolescentis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.149 overexpression in Escherichia coli BL21(DE3)RIPL Bifidobacterium adolescentis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.149 sitting drop method, crystallization of the ternary complex of NADH and cortisol bound to the inactive S181A mutant, the structure is determined at 2.2 A for the apoform and 2.0 A for the binary complex with NADH Bifidobacterium adolescentis

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.149 additional information important role of the 38-residue extended N-terminus in protein stability as deletion of the first 21 residues effectively abolishes enzyme activity with a 1.45 °C shift in Tm Bifidobacterium adolescentis
1.1.1.149 S181A inactive mutant enzyme Bifidobacterium adolescentis
1.1.1.149 S183A the kcat/Km-value for cortisol is 56% compared to wild-type enzyme Bifidobacterium adolescentis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.149 0.02407
-
 cortisol pH 5.0, temperature not specified in the publication Bifidobacterium adolescentis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.149 130500
-
 gel filtration Bifidobacterium adolescentis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.149 cortisol + NADH + H+ Bifidobacterium adolescentis
-
 20beta-hydroxycortisol + NAD+
-
 ?
1.1.1.149 cortisol + NADH + H+ Bifidobacterium adolescentis L2-32
-
 20beta-hydroxycortisol + NAD+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.149 Bifidobacterium adolescentis A7A7R9
-
-
1.1.1.149 Bifidobacterium adolescentis L2-32 A7A7R9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.149
-
 Bifidobacterium adolescentis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.149 11-deoxycortisol + NADH + H+ i.e. 4-pregnen-17,21-diol-3,20-dione, 66.03% compared to the activity with cortisol Bifidobacterium adolescentis ? + NAD+
-
 ?
1.1.1.149 11-deoxycortisol + NADH + H+ i.e. 4-pregnen-17,21-diol-3,20-dione, 66.03% compared to the activity with cortisol Bifidobacterium adolescentis L2-32 ? + NAD+
-
 ?
1.1.1.149 corticosterone + NADH + H+ i.e. 4-pregnen-11beta,21-diol-3,20-dione, 33.32% compared to the activity with cortisol Bifidobacterium adolescentis ? + NAD+
-
 ?
1.1.1.149 corticosterone + NADH + H+ i.e. 4-pregnen-11beta,21-diol-3,20-dione, 33.32% compared to the activity with cortisol Bifidobacterium adolescentis L2-32 ? + NAD+
-
 ?
1.1.1.149 cortisol + NADH + H+
-
 Bifidobacterium adolescentis 20beta-hydroxycortisol + NAD+
-
 ?
1.1.1.149 cortisol + NADH + H+ no activity with NADPH Bifidobacterium adolescentis 20beta-hydroxycortisol + NAD+
-
 ?
1.1.1.149 cortisol + NADH + H+
-
 Bifidobacterium adolescentis L2-32 20beta-hydroxycortisol + NAD+
-
 ?
1.1.1.149 cortisol + NADH + H+ no activity with NADPH Bifidobacterium adolescentis L2-32 20beta-hydroxycortisol + NAD+
-
 ?
1.1.1.149 tetrahydrocortisol + NADH + H+ i.e. 5beta-pregnan-3alpha,11beta,17,21-tetrol-20-one, 44.29% compared to the activity with cortisol Bifidobacterium adolescentis ? + NAD+
-
 ?
1.1.1.149 tetrahydrocortisol + NADH + H+ i.e. 5beta-pregnan-3alpha,11beta,17,21-tetrol-20-one, 44.29% compared to the activity with cortisol Bifidobacterium adolescentis L2-32 ? + NAD+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.149 homotetramer 4 * 31700, SDS-PAGE Bifidobacterium adolescentis

Synonyms

EC Number Synonyms Comment Organism
1.1.1.149 20beta-HSDH
-
 Bifidobacterium adolescentis
1.1.1.149 20beta-hydroxysteroid dehydrogenase
-
 Bifidobacterium adolescentis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.149 5
-
 recombinant enzyme, reductive direction Bifidobacterium adolescentis
1.1.1.149 5.5
-
 recombinant enzyme, oxidative direction Bifidobacterium adolescentis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.149 56
-
 transition temperature. Important role of the 38-residue extended N-terminus in protein stability as deletion of the first 21 residues effectively abolishes enzyme activity with a 1.45 °C shift in Tm Bifidobacterium adolescentis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.149 11.1
-
 cortisol pH 5.0, temperature not specified in the publication Bifidobacterium adolescentis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.149 NADH no activity with NADPH Bifidobacterium adolescentis

General Information

EC Number General Information Comment Organism
1.1.1.149 physiological function the enzyme regulates cortisol side-chain cleavage by reducing the C-20 carboxyl group on cortisol Bifidobacterium adolescentis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.149 461.1
-
 cortisol pH 5.0, temperature not specified in the publication Bifidobacterium adolescentis