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Literature summary for 5.4.99.64 extracted from

  • Weichler, M.T.; Kurteva-Yaneva, N.; Przybylski, D.; Schuster, J.; Mueller, R.H.; Harms, H.; Rohwerder, T.
    Thermophilic coenzyme B12-dependent acyl coenzyme A (CoA) mutase from Kyrpidia tusciae DSM 2912 preferentially catalyzes isomerization of (R)-3-hydroxybutyryl-CoA and 2-hydroxyisobutyryl-CoA (2015), Appl. Environ. Microbiol., 81, 4564-4572.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.4.99.64

Application

Application Comment Organism
synthesis introduction of a 2-hydroxyisobutyric acid synthesis route in Escherichia coli by concomitantly expressing the RCM genes and phaA and phaB, encoding beta-ketothiolase and NADP-dependent (R)-3-hydroxybutyryl-CoA dehydrogenase. A concentration of 3-hydroxybutyric acid of up to 17.7 mM can be obtained within 8 days of feeding the recombinant strain with gluconic acid as the main carbon source Kyrpidia tusciae DSM 2912

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Kyrpidia tusciae DSM 2912

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.087
-
 (R)-3-hydroxybutanoyl-CoA pH 7.8, 55°C Kyrpidia tusciae DSM 2912
0.09
-
 2-hydroxyisobutanoyl-CoA pH 7.8, 55°C Kyrpidia tusciae DSM 2912
0.313
-
 (S)-3-hydroxybutanoyl-CoA pH 7.8, 55°C Kyrpidia tusciae DSM 2912

Organism

Organism UniProt Comment Textmining
Kyrpidia tusciae DSM 2912 D5WTR7 and D5WTR8 D5WTR7 i.e. subunit RcmA, D5WTR8 i.e. subunit RcmB
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-3-hydroxybutanoyl-CoA catalytic efficiency for the (R)-enantiomer is about 11 times higher than for the (S)-enantiomer Kyrpidia tusciae DSM 2912 2-hydroxybutanoyl-CoA
-
 r
(S)-3-hydroxybutanoyl-CoA catalytic efficiency for the (R)-enantiomer is about 11 times higher than for the (S)-enantiomer Kyrpidia tusciae DSM 2912 2-hydroxybutanoyl-CoA
-
 r
2-hydroxyisobutanoyl-CoA
-
 Kyrpidia tusciae DSM 2912 (S)-3-hydroxyisobutanoyl-CoA + (R)-3-hydroxyisobutanoyl-CoA
-
 r
additional information (R)-3-hydroxybuytryl-CoA is converted 3.1 times faster than the (S)-enantiomer Kyrpidia tusciae DSM 2912 ?
-
 ?

Subunits

Subunits Comment Organism
multimer x * subunit RcmA + x * subunit RcmB Kyrpidia tusciae DSM 2912

Synonyms

Synonyms Comment Organism
Btus_0469 gene name, subunit RcmA Kyrpidia tusciae DSM 2912
Btus_0470 gene name, subunit RcmB Kyrpidia tusciae DSM 2912
RcmA
-
 Kyrpidia tusciae DSM 2912
RcmB
-
 Kyrpidia tusciae DSM 2912

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
 Kyrpidia tusciae DSM 2912

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30
-
 25% of maximum activity Kyrpidia tusciae DSM 2912
75
-
 55% of maximum activity Kyrpidia tusciae DSM 2912

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.6
-
 (S)-3-hydroxybutanoyl-CoA pH 7.8, 55°C Kyrpidia tusciae DSM 2912
3.7
-
 2-hydroxyisobutanoyl-CoA pH 7.8, 55°C Kyrpidia tusciae DSM 2912
4.9
-
 (R)-3-hydroxybutanoyl-CoA pH 7.8, 55°C Kyrpidia tusciae DSM 2912

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
-
 Kyrpidia tusciae DSM 2912

General Information

General Information Comment Organism
malfunction mutase from strain DSM 2912 catalyzes isomerization of (R)-3-hydroxybutanoyl-CoA about 7000 times more efficiently than the mutase from Aquincola tertiaricarbonis L108. The most striking structural difference between the two mutases, likely determining stereospecificity, is a replacement of active-site residue Asp found in strain L108 at position 117 with Val in the enzyme from strain DSM 2912, resulting in a reversed polarity at this binding site Kyrpidia tusciae DSM 2912

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5
-
 (S)-3-hydroxybutanoyl-CoA pH 7.8, 55°C Kyrpidia tusciae DSM 2912
40
-
 2-hydroxyisobutanoyl-CoA pH 7.8, 55°C Kyrpidia tusciae DSM 2912
56.7
-
 (R)-3-hydroxybutanoyl-CoA pH 7.8, 55°C Kyrpidia tusciae DSM 2912