BRENDA - Enzyme Database show
show all sequences of 5.4.99.64

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

Weichler, M.T.; Kurteva-Yaneva, N.; Przybylski, D.; Schuster, J.; Mueller, R.H.; Harms, H.; Rohwerder, T.; Appl. Environ. Microbiol. 81, 4564-4572 (2015)

Data extracted from this reference:

Application
Application
Commentary
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)
Commentary
Organism
expression in Escherichia coli
Kyrpidia tusciae DSM 2912
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.087
-
(R)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
0.09
-
2-hydroxyisobutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
0.313
-
(S)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Kyrpidia tusciae DSM 2912
D5WTR7 and D5WTR8
D5WTR7 i.e. subunit RcmA, D5WTR8 i.e. subunit RcmB
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-3-hydroxybutanoyl-CoA
catalytic efficiency for the (R)-enantiomer is about 11 times higher than for the (S)-enantiomer
737504
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
737504
Kyrpidia tusciae DSM 2912
2-hydroxybutanoyl-CoA
-
-
-
r
2-hydroxyisobutanoyl-CoA
-
737504
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
737504
Kyrpidia tusciae DSM 2912
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
multimer
x * subunit RcmA + x * subunit RcmB
Kyrpidia tusciae DSM 2912
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
55
-
-
Kyrpidia tusciae DSM 2912
Temperature Range [C]
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
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
Commentary
Organism
Structure
1.6
-
(S)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
3.7
-
2-hydroxyisobutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
4.9
-
(R)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.8
-
-
Kyrpidia tusciae DSM 2912
Application (protein specific)
Application
Commentary
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) (protein specific)
Commentary
Organism
expression in Escherichia coli
Kyrpidia tusciae DSM 2912
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.087
-
(R)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
0.09
-
2-hydroxyisobutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
0.313
-
(S)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-3-hydroxybutanoyl-CoA
catalytic efficiency for the (R)-enantiomer is about 11 times higher than for the (S)-enantiomer
737504
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
737504
Kyrpidia tusciae DSM 2912
2-hydroxybutanoyl-CoA
-
-
-
r
2-hydroxyisobutanoyl-CoA
-
737504
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
737504
Kyrpidia tusciae DSM 2912
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
multimer
x * subunit RcmA + x * subunit RcmB
Kyrpidia tusciae DSM 2912
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
55
-
-
Kyrpidia tusciae DSM 2912
Temperature Range [C] (protein specific)
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
30
-
25% of maximum activity
Kyrpidia tusciae DSM 2912
75
-
55% of maximum activity
Kyrpidia tusciae DSM 2912
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.6
-
(S)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
3.7
-
2-hydroxyisobutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
4.9
-
(R)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.8
-
-
Kyrpidia tusciae DSM 2912
General Information
General Information
Commentary
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
General Information (protein specific)
General Information
Commentary
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
Commentary
Organism
Structure
5
-
(S)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
40
-
2-hydroxyisobutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
56.7
-
(R)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
5
-
(S)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
40
-
2-hydroxyisobutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
56.7
-
(R)-3-hydroxybutanoyl-CoA
pH 7.8, 55C
Kyrpidia tusciae DSM 2912
Other publictions for EC 5.4.99.64
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
737504
Weichler
Thermophilic coenzyme B12-depe ...
Kyrpidia tusciae DSM 2912
Appl. Environ. Microbiol.
81
4564-4572
2015
-
1
1
-
-
-
-
3
-
-
-
-
-
7
-
-
-
-
-
-
-
-
4
1
1
2
-
3
1
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
4
1
1
2
-
3
1
-
-
-
-
1
1
-
3
3
738677
Kurteva-Yaneva
Structural basis of the stereo ...
Aquincola tertiaricarbonis
J. Biol. Chem.
290
9727-9737
2015
-
-
1
1
5
-
-
26
-
-
-
-
-
2
-
-
1
-
-
-
-
-
11
-
1
-
-
30
1
-
-
-
-
-
-
-
-
1
-
1
5
-
-
-
-
26
-
-
-
-
-
-
-
1
-
-
-
-
11
-
1
-
-
30
1
-
-
-
-
-
-
-
27
27
737532
Przybylski
Synthesis of the building bloc ...
Aquincola tertiaricarbonis, Aquincola tertiaricarbonis L108
Appl. Microbiol. Biotechnol.
97
8875-8885
2013
-
1
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
736409
Yaneva
Bacterial acyl-CoA mutase spec ...
Aquincola tertiaricarbonis, Aquincola tertiaricarbonis L108
J. Biol. Chem.
287
15502-15511
2012
-
-
1
-
3
-
-
7
-
-
2
2
-
4
-
-
1
-
-
-
-
-
12
1
1
1
-
7
1
1
-
1
-
-
-
-
-
1
1
-
3
-
-
-
-
7
-
-
2
2
-
-
-
1
-
-
-
-
12
1
1
1
-
7
1
1
-
-
-
1
1
-
7
7