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Literature summary for 1.1.1.30 extracted from

  • Paithankar, K.S.; Feller, C.; Kuettner, E.B.; Keim, A.; Grunow, M.; Sträter, N.
    Cosubstrate-induced dynamics of D-3-hydroxybutyrate dehydrogenase from Pseudomonas putida (2007), FEBS J., 274, 5767-5779.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
enzyme expression in Escherichia coli strain XL-1 Blue Pseudomonas putida
expressed in Escherichia coli XL1-Blue cells Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapour diffusion method with 17-20% polyethylene glycol 1500, 0.1 M Tris-HCl, pH 7.1, 0.2 mM CaCl2 and 10 mM acetoacetate Pseudomonas putida
recombinant enzyme, hanging drop vapor diffusion method, 0.003 ml of HBDH, 10 mg/ml, is mixed with an equal volume of crystallization buffer containing 17-20% PEG 1500, 0.1 M Tris-HCl, pH 7.1, 0.2 mM CaCl2, and 10 mM acetoacetate, room temperature/22°C, three different crystal forms, X-ray diffraction structure determination and analysis at resolutions between 1.9 and 2.1 A Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
H141A strongly decreased activity Pseudomonas putida
L149A inactive Pseudomonas putida
Q133A decreased activity Pseudomonas putida
Q91A decreased activity Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.6
-
(R)-3-hydroxybutanoate wild type enzyme Pseudomonas putida
4
-
(R)-3-hydroxybutanoate mutant enzyme H141A Pseudomonas putida
51
-
(R)-3-hydroxybutanoate mutant enzyme Q91A Pseudomonas putida
70
-
(R)-3-hydroxybutanoate mutant enzyme Q133A Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(R)-3-hydroxybutanoate + NAD+ Pseudomonas putida
-
acetoacetate + NADH
-
r

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
-
-
Pseudomonas putida Q9AE70
-
-

Purification (Commentary)

Purification (Comment) Organism
charge-controlled hydrophobic chromatography and Sephadex G-100 gel filtration Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-3-hydroxybutanoate + NAD+
-
Pseudomonas putida acetoacetate + NADH
-
r
(R)-3-hydroxybutanoate + NAD+ model of the binding mode of the substrate D-3-hydroxybutyrate, Gln193 is the only residue of the substrate-binding loop that interacts directly with the substrate, NAD+ binding increases the flexibility of the substrate-binding loop and shifts the equilibrium between the open and closed forms towards the closed form, overview Pseudomonas putida acetoacetate + NADH
-
r
(R)-3-hydroxybutanoate + NAD+
-
Pseudomonas putida acetoacetate + NADH + H+
-
r

Subunits

Subunits Comment Organism
tetramer x-ray crystallography Pseudomonas putida

Synonyms

Synonyms Comment Organism
D-3-hydroxybutyrate dehydrogenase
-
Pseudomonas putida
HBDH
-
Pseudomonas putida
More the enzyme belongs to the family of short-chain dehydrogenases/reductases Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
13
-
(R)-3-hydroxybutanoate mutant enzyme H141A Pseudomonas putida
215
-
(R)-3-hydroxybutanoate mutant enzyme Q133A Pseudomonas putida
411
-
(R)-3-hydroxybutanoate mutant enzyme Q91A Pseudomonas putida
432
-
(R)-3-hydroxybutanoate wild type enzyme Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Pseudomonas putida
NAD+ the substrate-binding loop, residues 187-210, is partially disordered in several subunits, in both the presence and absence of NAD+, closed conformation in the complex structure with NAD+, interactions of Val185, Thr187 and Leu189 with the cosubstrate induced the conformational change from open to closed, NAD+ binding increases the flexibility of the substrate-binding loop and shifts the equilibrium between the open and closed forms towards the closed form Pseudomonas putida
NADH
-
Pseudomonas putida