Literature summary for 5.4.99.64 extracted from

Yaneva, N.; Schuster, J.; Schafer, F.; Lede, V.; Przybylski, D.; Paproth, T.; Harms, H.; Muller, R.H.; Rohwerder, T.
Bacterial acyl-CoA mutase specifically catalyzes coenzyme B12-dependent isomerization of 2-hydroxyisobutyryl-CoA and (S)-3-hydroxybutyryl-CoA (2012), J. Biol. Chem., 287, 15502-15511.

Search for more literature for 5.4.99.64

Cloned(Commentary)

Cloned (Comment)	Organism
wild-type mutase genes are cloned in Escherichia coli strains	Aquincola tertiaricarbonis

Protein Variants

Protein Variants	Comment	Organism
I90F	complete loss of enzyme activity	Aquincola tertiaricarbonis
I90V	significant activities are only obtained with (S)-3-hydroxybutanoyl- and 2-hydroxyisobutanoyl-CoA, showing less than 20% of the wild-type rates. The substitution I90V results in Km values close to 2 mM and a nearly 100fold diminution of the catalytic efficiency with (S)-3-hydroxybuytryl-CoA	Aquincola tertiaricarbonis
I90Y	complete loss of enzyme activity	Aquincola tertiaricarbonis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.104	-	2-hydroxy-2-methylpropanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
0.128	-	(S)-3-hydroxybutanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
0.55	-	isobutanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
1.66	-	(R)-3-hydroxybutanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
1.76	-	(S)-3-hydroxybutanoyl-CoA	pH 6.6, 30°C, mutant enzyme I90V	Aquincola tertiaricarbonis
1.84	-	2-hydroxy-2-methylpropanoyl-CoA	pH 6.6, 30°C, mutant enzyme I90V	Aquincola tertiaricarbonis
3.34	-	butanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
14515	-	1 * 63414 + 1 * 14515, calculated from sequence, the enzyme consists of a large acyl-CoA-binding and a small B12-binding subunit, HcmA and HcmB, respectively	Aquincola tertiaricarbonis
63414	-	1 * 63414 + 1 * 14515, calculated from sequence, the enzyme consists of a large acyl-CoA-binding and a small B12-binding subunit, HcmA and HcmB, respectively	Aquincola tertiaricarbonis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2-hydroxy-2-methylpropanoyl-CoA	Aquincola tertiaricarbonis	the enzym plays a central role in the degradation of assorted substrates containing a tert-butyl moiety	(S)-3-hydroxybutanoyl-CoA	-	r
2-hydroxy-2-methylpropanoyl-CoA	Aquincola tertiaricarbonis L108	the enzym plays a central role in the degradation of assorted substrates containing a tert-butyl moiety	(S)-3-hydroxybutanoyl-CoA	-	r

Organism

Organism	UniProt	Comment	Textmining
Aquincola tertiaricarbonis	I3VE77 and I3VE74	I3VE77: large subunit hcmA, I3VE74: small subunit hcmB	-
Aquincola tertiaricarbonis L108	I3VE77 and I3VE74	I3VE77: large subunit hcmA, I3VE74: small subunit hcmB	-

Purification (Commentary)

Purification (Comment)	Organism
-	Aquincola tertiaricarbonis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(R)-3-hydroxybutanoyl-CoA	2-hydroxy-2-methylpropanoyl i.e. 2-hydroxyisobutanoyl-CoA. 2-Hydroxyisobutanoyl-CoA is predominantly converted to (S)-3-hydroxybutanoyl-CoA, and only minor amounts of about 20% of the (R)-enantiomer are obtained	Aquincola tertiaricarbonis	2-hydroxy-2-methylpropanoyl-CoA	-	r
(R)-3-hydroxybutanoyl-CoA	2-hydroxy-2-methylpropanoyl i.e. 2-hydroxyisobutanoyl-CoA. 2-Hydroxyisobutanoyl-CoA is predominantly converted to (S)-3-hydroxybutanoyl-CoA, and only minor amounts of about 20% of the (R)-enantiomer are obtained	Aquincola tertiaricarbonis L108	2-hydroxy-2-methylpropanoyl-CoA	-	r
(S)-3-hydroxybutanoyl-CoA	-	Aquincola tertiaricarbonis	2-hydroxy-2-methylpropanoyl-CoA	-	r
(S)-3-hydroxybutanoyl-CoA	-	Aquincola tertiaricarbonis L108	2-hydroxy-2-methylpropanoyl-CoA	-	r
2-hydroxy-2-methylpropanoyl-CoA	the enzym plays a central role in the degradation of assorted substrates containing a tert-butyl moiety	Aquincola tertiaricarbonis	(S)-3-hydroxybutanoyl-CoA	-	r
2-hydroxy-2-methylpropanoyl-CoA	2-hydroxy-2-methylpropanoyl i.e. 2-hydroxyisobutanoyl-CoA. 2-Hydroxyisobutanoyl-CoA is predominantly converted to (S)-3-hydroxybutanoyl-CoA, and only minor amounts of about 20% of the (R)-enantiomer are obtained, indicating stereospecific catalysis. The enzyme uses radical chemistry to rearrange the positions of both a methyl group and a hydroxyl group. Compared with Vmax obtained with 2-hydroxyisobutanoyl-CoA, rates with (S)-3-hydroxybutanoyl-CoA are nearly four times higher	Aquincola tertiaricarbonis	(S)-3-hydroxybutanoyl-CoA	-	r
2-hydroxy-2-methylpropanoyl-CoA	the enzym plays a central role in the degradation of assorted substrates containing a tert-butyl moiety	Aquincola tertiaricarbonis L108	(S)-3-hydroxybutanoyl-CoA	-	r
2-hydroxy-2-methylpropanoyl-CoA	2-hydroxy-2-methylpropanoyl i.e. 2-hydroxyisobutanoyl-CoA. 2-Hydroxyisobutanoyl-CoA is predominantly converted to (S)-3-hydroxybutanoyl-CoA, and only minor amounts of about 20% of the (R)-enantiomer are obtained, indicating stereospecific catalysis. The enzyme uses radical chemistry to rearrange the positions of both a methyl group and a hydroxyl group. Compared with Vmax obtained with 2-hydroxyisobutanoyl-CoA, rates with (S)-3-hydroxybutanoyl-CoA are nearly four times higher	Aquincola tertiaricarbonis L108	(S)-3-hydroxybutanoyl-CoA	-	r
butanoyl-CoA	low rearrangement activity, corresponding to approximately 2% of the (S)-3-hydroxybutanoyl-CoA conversion rate	Aquincola tertiaricarbonis	isobutanoyl-CoA	-	r
isobutanoyl-CoA	low rearrangement activity, corresponding to approximately 0.5% of the (S)-3-hydroxybutanoyl-CoA conversion rate	Aquincola tertiaricarbonis	butanoyl-CoA	-	r
additional information	the enzyme does not show detectable conversion of methylmalonyl-CoA and succinyl-CoA	Aquincola tertiaricarbonis	?	-	?
additional information	the enzyme does not show detectable conversion of methylmalonyl-CoA and succinyl-CoA	Aquincola tertiaricarbonis L108	?	-	?

Subunits

Subunits	Comment	Organism
dimer	1 * 63414 + 1 * 14515, calculated from sequence, the enzyme consists of a large acyl-CoA-binding and a small B12-binding subunit, HcmA and HcmB, respectively	Aquincola tertiaricarbonis

Synonyms

Synonyms	Comment	Organism
2-hydroxyisobutyryl-CoA mutase	-	Aquincola tertiaricarbonis
hcmAB	-	Aquincola tertiaricarbonis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	-	Aquincola tertiaricarbonis

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
25	35	25°C: about 50% pf maxim al activity, 35°C: about 45% of maximal activity	Aquincola tertiaricarbonis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.001	-	isobutanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
0.0033	-	butanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
0.0033	-	(R)-3-hydroxybutanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
0.02	-	2-hydroxy-2-methylpropanoyl-CoA	pH 6.6, 30°C, mutant enzyme I90V	Aquincola tertiaricarbonis
0.033	-	(S)-3-hydroxybutanoyl-CoA	pH 6.6, 30°C, mutant enzyme I90V	Aquincola tertiaricarbonis
0.05	-	2-hydroxy-2-methylpropanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
0.2	-	(S)-3-hydroxybutanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.6	-	-	Aquincola tertiaricarbonis

pH Range

pH Minimum	pH Maximum	Comment	Organism
6.2	7	pH 6.2: about 60% of maximal activity, pH 7.0: about 60% of maximal activity	Aquincola tertiaricarbonis

Cofactor

Cofactor	Comment	Organism	Structure
Cobalamin	the smaller one of the two subunits (hcmB) contains a cobalamin cofactor	Aquincola tertiaricarbonis

General Information

General Information	Comment	Organism
metabolism	the enzym plays a central role in the degradation of assorted substrates containing a tert-butyl moiety	Aquincola tertiaricarbonis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.0018	-	isobutanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
0.00199	-	(R)-3-hydroxybutanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
0.0099	-	butanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
0.011	-	2-hydroxy-2-methylpropanoyl-CoA	pH 6.6, 30°C, mutant enzyme I90V	Aquincola tertiaricarbonis
0.019	-	(S)-3-hydroxybutanoyl-CoA	pH 6.6, 30°C, mutant enzyme I90V	Aquincola tertiaricarbonis
0.48	-	2-hydroxy-2-methylpropanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis
1.56	-	(S)-3-hydroxybutanoyl-CoA	pH 6.6, 30°C, wild-type enzyme	Aquincola tertiaricarbonis

Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.

Release 2023.1 (January 2023)