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Literature summary extracted from

  • Hedl, M.; Sutherlin, A.; Wilding, E.I.; Mazzulla, M.; McDevitt, D.; Lane, P.; Burgner, J.W.; Lehnbeuter, K.R.; Stauffacher, C.V.; Gwynn, M.N.; Rodwell, V.W.
    Enterococcus faecalis acetoacetyl-coenzyme A thiolase/3-hydroxy-3-methylglutaryl-coenzyme A reductase, a dual-function protein of isopentenyl diphosphate biosynthesis (2002), J. Bacteriol., 184, 2116-2122.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.34 expression in Escherichia coli strain DH5alpha as His-tagged enzyme Enterococcus faecalis
2.3.1.9 expression in Escherichia coli Enterococcus faecalis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.1.9 CoA
-
Enterococcus faecalis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.34 0.02
-
(S)-3-hydroxy-3-methylglutaryl-CoA pH 6.5, 37°C Enterococcus faecalis
1.1.1.34 0.03
-
NADPH pH 6.5, 37°C, (S)-3-hydroxy-3-methylglutaryl-CoA reduction Enterococcus faecalis
1.1.1.34 0.22
-
CoA pH 6.5, 37°C, mevaldehyde oxidation Enterococcus faecalis
1.1.1.34 0.23
-
CoA pH 6.5, 37°C, mevalonate oxidation Enterococcus faecalis
1.1.1.34 0.25
-
NADPH pH 6.5, 37°C, mevaldehyde reduction Enterococcus faecalis
1.1.1.34 0.25
-
NADP+ pH 6.5, 37°C, mevalonate oxidation Enterococcus faecalis
1.1.1.34 0.66
-
mevaldehyde pH 6.5, 37°C, oxidation Enterococcus faecalis
1.1.1.34 0.67
-
NADP+ pH 6.5, 37°C, mevaldehyde oxidation Enterococcus faecalis
1.1.1.34 1
-
(R)-mevalonate pH 6.5, 37°C, oxidation Enterococcus faecalis
1.1.1.34 3.8
-
mevaldehyde pH 6.5, 37°C, reduction Enterococcus faecalis
2.3.1.9 0.01
-
CoA thiolysis Enterococcus faecalis
2.3.1.9 0.088
-
acetoacetyl-CoA thiolysis Enterococcus faecalis
2.3.1.9 0.6
-
acetyl-CoA synthesis Enterococcus faecalis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.1.9 86496
-
x * 86496, deduced from nucleotide sequence Enterococcus faecalis
2.3.1.9 88712
-
x * 88712, MALDI-mass spectrometry Enterococcus faecalis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.34 (S)-3-hydroxy-3-methylglutaryl-CoA + 2 NADPH + 2 H+ Enterococcus faecalis involved in isopentenyl diphosphate biosynthesis, mevalonate pathway overview (R)-mevalonate + CoA + 2 NADP+
-
r
1.1.1.34 acetyl-CoA Enterococcus faecalis
-
acetoacetyl-CoA + CoA
-
r
2.3.1.9 CoA + acetoacetyl-CoA Enterococcus faecalis
-
acetyl-CoA + acetyl-CoA
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.34 Enterococcus faecalis
-
dual-function enzyme with acetoacetyl-coenzyme A thiolase and 3-hydroxy-3-methylglutaryl-coenzyme A reductase activities
-
2.3.1.9 Enterococcus faecalis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.34 recombinant His-tagged enzyme from Escherichia coli by nickel-affinity chromatography Enterococcus faecalis
2.3.1.9 recombinant His-tagged thiolase, Ni-affinity column Enterococcus faecalis

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.34 (R)-mevalonate + CoA + 2 NADP+ = (S)-3-hydroxy-3-methylglutaryl-CoA + 2 NADPH + 2 H+ via mevaldehyde as an intermediate product Enterococcus faecalis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.34 (S)-3-hydroxy-3-methylglutaryl-CoA + 2 NADPH + 2 H+ involved in isopentenyl diphosphate biosynthesis, mevalonate pathway overview Enterococcus faecalis (R)-mevalonate + CoA + 2 NADP+
-
r
1.1.1.34 (S)-3-hydroxy-3-methylglutaryl-CoA + 2 NADPH + 2 H+ overall reaction Enterococcus faecalis (R)-mevalonate + CoA + 2 NADP+
-
r
1.1.1.34 (S)-3-hydroxy-3-methylglutaryl-CoA + NADPH + H+ first step reaction Enterococcus faecalis mevaldehyde + CoA + NADP+
-
r
1.1.1.34 acetyl-CoA
-
Enterococcus faecalis acetoacetyl-CoA + CoA
-
r
1.1.1.34 mevaldehyde + NADPH + H+ second step reaction Enterococcus faecalis (R)-mevalonate + NADP+
-
r
2.3.1.9 CoA + acetoacetyl-CoA
-
Enterococcus faecalis acetyl-CoA + acetyl-CoA
-
?
2.3.1.9 CoA + acetoacetyl-CoA enzyme has both thiolase and 3-hydroxy-3-methylglutaryl-CoA reductase activity Enterococcus faecalis acetyl-CoA + acetyl-CoA
-
r

Subunits

EC Number Subunits Comment Organism
2.3.1.9 ? x * 88712, MALDI-mass spectrometry Enterococcus faecalis
2.3.1.9 ? x * 86496, deduced from nucleotide sequence Enterococcus faecalis

Synonyms

EC Number Synonyms Comment Organism
1.1.1.34 acetoacetyl-coenzyme A thiolase/3-hydroxy-3-methylglutaryl-coenzyme A reductase
-
Enterococcus faecalis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.34 37
-
about, acetoacetyl-coenzyme A synthesis and thiolysis Enterococcus faecalis
2.3.1.9 37
-
acetoacetyl-CoA synthesis and thiolysis Enterococcus faecalis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.34 6.5
-
(S)-3-hydroxy-3-methylglutaryl-CoA reduction Enterococcus faecalis
1.1.1.34 7
-
(R)-mevalonate oxidation Enterococcus faecalis
1.1.1.34 9.5
-
acetoacetyl-coenzyme A thiolysis Enterococcus faecalis
1.1.1.34 10.5
-
acetoacetyl-coenzyme A synthesis Enterococcus faecalis
2.3.1.9 9.5
-
thiolysis Enterococcus faecalis
2.3.1.9 10.5
-
synthesis Enterococcus faecalis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.3.1.9 0.12
-
CoA
-
Enterococcus faecalis