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Literature summary for 2.8.3.24 extracted from

  • Kim, J.; Darley, D.; Selmer, T.; Buckel, W.
    Characterization of (R)-2-hydroxyisocaproate dehydrogenase and a family III coenzyme A transferase involved in reduction of L-leucine to isocaproate by Clostridium difficile (2006), Appl. Environ. Microbiol., 72, 6062-6069.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.8.3.24

Cloned(Commentary)

Cloned (Comment) Organism
overexpresssion in Escherichia coli Clostridioides difficile

Protein Variants

Protein Variants Comment Organism
D171A the mutant enzyme is more than 2000-fold less active than the wild-type enzyme Clostridioides difficile
D171N the mutant enzyme is more than 2000-fold less active than the wild-type enzyme Clostridioides difficile

Inhibitors

Inhibitors Comment Organism Structure
hydroxylamine after incubation with 200 mM hydroxylamine at pH 7.0 in the presence of 0.2 mM (R)-2-hydroxyisocaproyl-CoA and removal of the small molecules by gel filtration, the transferase sample shows 94% inactivation, while no inactivation is observed under the same conditions in the absence of (R)-2-hydroxyisocaproyl-CoA Clostridioides difficile
NaBH4 after incubation with 20 mM NaBH4 at pH 7.0 in the presence of 0.2 mM (R)-2-hydroxyisocaproyl-CoA and removal of the small molecules by gel filtration, the transferase sample shows 88% inactivation, while no inactivation is observed under the same conditions in the absence of (R)-2-hydroxyisocaproyl-CoA Clostridioides difficile

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
43000
-
 2 * 43000, C-terminal Strep tag II fusion protein, SDS-PAGE Clostridioides difficile
86000
-
 C-terminal Strep tag II fusion protein, gel filtration Clostridioides difficile

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4-methylpentanoyl-CoA + (R)-2-hydroxy-4-methylpentanoate Clostridioides difficile the enzyme participates in an L-leucine fermentation pathway 4-methylpentanoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?
4-methylpentanoyl-CoA + (R)-2-hydroxy-4-methylpentanoate Clostridioides difficile DSM 1296T the enzyme participates in an L-leucine fermentation pathway 4-methylpentanoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?

Organism

Organism UniProt Comment Textmining
Clostridioides difficile
-
-
-
Clostridioides difficile DSM 1296T
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Clostridioides difficile

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylpentanoyl-CoA + (R)-2-hydroxy-4-methylpentanoate the enzyme participates in an L-leucine fermentation pathway Clostridioides difficile 4-methylpentanoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?
4-methylpentanoyl-CoA + (R)-2-hydroxy-4-methylpentanoate the reaction proceeds via formation of a covalent anhydride intermediate between a conserved aspartate residue and the acyl group of the CoA thioester substrate. Other CoA thioesters, such as acetyl-CoA or butyryl-CoA, are not accepted as substrates. Unexpectedly, CoA transfers from chemically synthesized (E)-2-isocaprenoyl-CoA to (R)-2-hydroxyisocaproate or to isocaproate cand not be observed Clostridioides difficile 4-methylpentanoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?
4-methylpentanoyl-CoA + (R)-2-hydroxy-4-methylpentanoate the enzyme participates in an L-leucine fermentation pathway Clostridioides difficile DSM 1296T 4-methylpentanoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?
4-methylpentanoyl-CoA + (R)-2-hydroxy-4-methylpentanoate the reaction proceeds via formation of a covalent anhydride intermediate between a conserved aspartate residue and the acyl group of the CoA thioester substrate. Other CoA thioesters, such as acetyl-CoA or butyryl-CoA, are not accepted as substrates. Unexpectedly, CoA transfers from chemically synthesized (E)-2-isocaprenoyl-CoA to (R)-2-hydroxyisocaproate or to isocaproate cand not be observed Clostridioides difficile DSM 1296T 4-methylpentanoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 43000, C-terminal Strep tag II fusion protein, SDS-PAGE Clostridioides difficile

Synonyms

Synonyms Comment Organism
(R)-2-hydroxyisocaproate CoA transferase
-
 Clostridioides difficile
HadA
-
 Clostridioides difficile

General Information

General Information Comment Organism
metabolism the enzyme participates in an L-leucine fermentation pathway Clostridioides difficile