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Literature summary for 1.1.1.149 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

Application Comment Organism
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)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
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
S181A inactive mutant enzyme Bifidobacterium adolescentis
S183A the kcat/Km-value for cortisol is 56% compared to wild-type enzyme Bifidobacterium adolescentis

KM Value [mM]

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Bifidobacterium adolescentis A7A7R9
-
-
Bifidobacterium adolescentis L2-32 A7A7R9
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Bifidobacterium adolescentis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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+
-
?
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+
-
?
corticosterone + NADH + H+ i.e. 4-pregnen-11beta,21-diol-3,20-dione, 33.32% compared to the activity with cortisol Bifidobacterium adolescentis ? + NAD+
-
?
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+
-
?
cortisol + NADH + H+
-
Bifidobacterium adolescentis 20beta-hydroxycortisol + NAD+
-
?
cortisol + NADH + H+ no activity with NADPH Bifidobacterium adolescentis 20beta-hydroxycortisol + NAD+
-
?
cortisol + NADH + H+
-
Bifidobacterium adolescentis L2-32 20beta-hydroxycortisol + NAD+
-
?
cortisol + NADH + H+ no activity with NADPH Bifidobacterium adolescentis L2-32 20beta-hydroxycortisol + NAD+
-
?
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+
-
?
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

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

Synonyms

Synonyms Comment Organism
20beta-HSDH
-
Bifidobacterium adolescentis
20beta-hydroxysteroid dehydrogenase
-
Bifidobacterium adolescentis

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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]

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

Cofactor

Cofactor Comment Organism Structure
NADH no activity with NADPH Bifidobacterium adolescentis

General Information

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

kcat/KM [mM/s]

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