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Literature summary for 6.2.1.1 extracted from

  • Ingram-Smith, C.; Woods, B.I.; Smith, K.S.
    Characterization of the acyl substrate binding pocket of acetyl-CoA synthetase (2006), Biochemistry, 45, 11482-11490.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Methanothermobacter thermautotrophicus

Protein Variants

Protein Variants Comment Organism
I312A kcat/Km for acetate is 15fold lower than wild-type value, kcat/Km for propionate is 5.2fold lower than wild-type value. Mutant enzyme shows activity with butyrate Methanothermobacter thermautotrophicus
T313K Km for acetate is 170fold higher than wild-type value, kcat for acetate is 34fold lower than wild-type value Methanothermobacter thermautotrophicus
T313V kcat/Km for acetate is 2.5fold lower than wild-type value, kcat/Km for propionate is identical to wild-type value Methanothermobacter thermautotrophicus
V388A kcat/Km for acetate is 6.9fold lower than wild-type value, kcat/Km for propionate is 2.5fold higher than wild-type value, mutant enzyme shows activity with wild-type enzyme Methanothermobacter thermautotrophicus
V388G kcat/Km for acetate is 93fold lower than wild-type value, kcat/Km for propionate is 26fold lower than wild-type value Methanothermobacter thermautotrophicus
W416G kcat/Km for acetate is 71.5fold lower than wild-type value, kcat/Km for propionate is 13fold lower than wild-type value, mutant enzyme shows activity with: butyrate, valerate, hexanoate, heptanoate, octanoate, 4-methylvalerate and 3-methylvalerate Methanothermobacter thermautotrophicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
CoA 65°C, mutant enzyme I312A Methanothermobacter thermautotrophicus
0.18
-
CoA 65°C, mutant enzyme V388G Methanothermobacter thermautotrophicus
0.19
-
CoA 65°C, wild-type enzyme Methanothermobacter thermautotrophicus
0.35
-
CoA 65°C, mutant enzyme V388A Methanothermobacter thermautotrophicus
0.43
-
CoA 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
0.46
-
Butyrate 65°C, mutant enzyme T313V Methanothermobacter thermautotrophicus
0.65
-
ATP 65°C, mutant enzyme T313V Methanothermobacter thermautotrophicus
0.7
-
CoA 65°C, mutant enzyme T313V Methanothermobacter thermautotrophicus
2.3
-
ATP 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
2.6
-
ATP 65°C, mutant enzyme V388A Methanothermobacter thermautotrophicus
3.3
-
ATP 65°C, wild-type enzyme Methanothermobacter thermautotrophicus
3.4
-
ATP 65°C, mutant enzyme T313K Methanothermobacter thermautotrophicus
3.5
-
acetate 65°C, wild-type enzyme Methanothermobacter thermautotrophicus
4
-
ATP 65°C, mutant enzyme V388G Methanothermobacter thermautotrophicus
4.1
-
propionate 65°C, mutant enzyme V388A Methanothermobacter thermautotrophicus
4.5
-
propionate 65°C, mutant enzyme T313V Methanothermobacter thermautotrophicus
6.1
-
hexanoate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
6.6
-
ATP 65°C, mutant enzyme I312A Methanothermobacter thermautotrophicus
7.6
-
4-methylvalerate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
10.2
-
Heptanoate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
11.1
-
valerate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
13.2
-
acetate 65°C, mutant enzyme V388A Methanothermobacter thermautotrophicus
18.1
-
Octanoate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
24.6
-
acetate 65°C, mutant enzyme I312A Methanothermobacter thermautotrophicus
29.2
-
2-methylvalerate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
36.5
-
propionate 65°C, wild-type enzyme Methanothermobacter thermautotrophicus
39.2
-
Butyrate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
128.6
-
propionate 65°C, mutant enzyme V388G Methanothermobacter thermautotrophicus
132.1
-
acetate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
151.9
-
Butyrate 65°C, mutant enzyme V388A Methanothermobacter thermautotrophicus
164.4
-
acetate 65°C, mutant enzyme V388G Methanothermobacter thermautotrophicus
188.8
-
propionate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
596
-
acetate 65°C, mutant enzyme T313K Methanothermobacter thermautotrophicus

Organism

Organism UniProt Comment Textmining
Methanothermobacter thermautotrophicus
-
-
-
Methanothermobacter thermautotrophicus Z245
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Methanothermobacter thermautotrophicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 2-methylvalerate + CoA
-
Methanothermobacter thermautotrophicus AMP + diphosphate + 2-methylvaleryl-CoA
-
?
ATP + 3-methylvalerate + CoA mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme Methanothermobacter thermautotrophicus AMP + diphosphate + 3-methylvaleryl-CoA
-
?
ATP + 4-methylvalerate + CoA mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme Methanothermobacter thermautotrophicus AMP + diphosphate + 4-methylvaleryl-CoA
-
?
ATP + acetate + CoA
-
Methanothermobacter thermautotrophicus AMP + diphosphate + acetyl-CoA
-
?
ATP + acetate + CoA ATP in form of MgATP2- Methanothermobacter thermautotrophicus AMP + diphosphate + acetyl-CoA
-
?
ATP + acetate + CoA
-
Methanothermobacter thermautotrophicus Z245 AMP + diphosphate + acetyl-CoA
-
?
ATP + acetate + CoA ATP in form of MgATP2- Methanothermobacter thermautotrophicus Z245 AMP + diphosphate + acetyl-CoA
-
?
ATP + butyrate + CoA mutant enzymes I312A and W416G catalyzes the reaction, no activity with wild-type enzyme Methanothermobacter thermautotrophicus AMP + diphosphate + butyryl-CoA
-
?
ATP + butyrate + CoA mutant enzymes I312A and W416G catalyzes the reaction, no activity with wild-type enzyme Methanothermobacter thermautotrophicus Z245 AMP + diphosphate + butyryl-CoA
-
?
ATP + heptanoate + CoA mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme Methanothermobacter thermautotrophicus AMP + diphosphate + heptanoyl-CoA
-
?
ATP + hexanoate + CoA mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme Methanothermobacter thermautotrophicus AMP + diphosphate + hexanoyl-CoA
-
?
ATP + hexanoate + CoA mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme Methanothermobacter thermautotrophicus Z245 AMP + diphosphate + hexanoyl-CoA
-
?
ATP + octanoate + CoA mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme Methanothermobacter thermautotrophicus AMP + diphosphate + octanoyl-CoA
-
?
ATP + propionate + CoA
-
Methanothermobacter thermautotrophicus AMP + diphosphate + propionyl-CoA
-
?
ATP + propionate + CoA
-
Methanothermobacter thermautotrophicus Z245 AMP + diphosphate + propionyl-CoA
-
?
ATP + valerate + CoA mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme Methanothermobacter thermautotrophicus AMP + diphosphate + valeryl-CoA
-
?

Synonyms

Synonyms Comment Organism
MT-ACS1
-
Methanothermobacter thermautotrophicus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.67
-
2-methylvalerate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
1.56
-
Butyrate 65°C, mutant enzyme V388A Methanothermobacter thermautotrophicus
1.9
-
acetate 65°C, mutant enzyme T313K Methanothermobacter thermautotrophicus
3.3
-
Octanoate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
3.4
-
Butyrate 65°C, mutant enzyme T313V Methanothermobacter thermautotrophicus
5.9
-
propionate 65°C, mutant enzyme T313V Methanothermobacter thermautotrophicus
5.9
-
hexanoate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
6
-
propionate 65°C, mutant enzyme V388G Methanothermobacter thermautotrophicus
7.3
-
Heptanoate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
10.8
-
Butyrate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
10.9
-
4-methylvalerate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
11.9
-
valerate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
13.2
-
propionate 65°C, mutant enzyme V388A Methanothermobacter thermautotrophicus
19.2
-
propionate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
30.4
-
acetate 65°C, mutant enzyme I312A Methanothermobacter thermautotrophicus
31.9
-
acetate 65°C, mutant enzyme V388G Methanothermobacter thermautotrophicus
33.7
-
acetate 65°C, mutant enzyme W416G Methanothermobacter thermautotrophicus
36.2
-
acetate 65°C, mutant enzyme V388A Methanothermobacter thermautotrophicus
46.3
-
propionate 65°C, wild-type enzyme Methanothermobacter thermautotrophicus
65.4
-
acetate 65°C, wild-type enzyme Methanothermobacter thermautotrophicus