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Literature summary extracted from

  • Altincicek, B.; Duin, E.C.; Reichenberg, A.; Hedderich, R.; Kollas, A.K.; Hintz, M.; Wagner, S.; Wiesner, J.; Beck, E.; Jomaa, H.
    LytB protein catalyzes the terminal step of the 2-C-methyl-D-erythritol-4-phosphate pathway of isoprenoid biosynthesis (2002), FEBS Lett., 532, 437-440.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.17.7.4 gene lytB, expression as His-tagged enzyme in Escherichia coli strain XL1-Blue Aquifex aeolicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.17.7.4 0.59
-
(E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate recombinant enzyme, pH 7.5, 60°C Aquifex aeolicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.17.7.4 (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ Aquifex aeolicus
-
dimethylallyl diphosphate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
r
1.17.7.4 isopentenyl diphosphate + NAD(P)+ + H2O Aquifex aeolicus part of isoprenoid biosynthesis (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H
-
r
1.17.7.4 isopentenyl diphosphate + NAD(P)+ + H2O Aquifex aeolicus 2-C-methyl-D-erithritol-4-phosphate pathway (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H
-
r

Organism

EC Number Organism UniProt Comment Textmining
1.17.7.4 Aquifex aeolicus O67625 gene lytB
-

Oxidation Stability

EC Number Oxidation Stability Organism
1.17.7.4 highly sensitive to O2, 21% remaining activity after incubation in air for 10 min Aquifex aeolicus

Purification (Commentary)

EC Number Purification (Comment) Organism
1.17.7.4 recombinant His-tagged protein from Escherichia coli strain Xl1-Blue, to 95% homogeneity Aquifex aeolicus

Reaction

EC Number Reaction Comment Organism Reaction ID
1.17.7.4 isopentenyl diphosphate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O = (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ reaction mechanism Aquifex aeolicus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.17.7.4 6.6
-
recombinant enzyme Aquifex aeolicus

Storage Stability

EC Number Storage Stability Organism
1.17.7.4 -20°C, purified enzyme, 20 mM Tris-HCl, pH 8.0, 95% remaining activity after 2 weeks Aquifex aeolicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.17.7.4 (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+
-
Aquifex aeolicus isopentenyl diphosphate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O producing a 5:1 mixture of isopentenyl diphosphate and dimethyallyl diphosphate r
1.17.7.4 (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+
-
Aquifex aeolicus dimethylallyl diphosphate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
r
1.17.7.4 (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+
-
Aquifex aeolicus dimethylallyl diphosphate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O producing a 5:1 mixture of isopentenyl diphosphate and dimethyallyl diphosphate r
1.17.7.4 isopentenyl diphosphate + NAD(P)+ + H2O part of isoprenoid biosynthesis Aquifex aeolicus (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H
-
r
1.17.7.4 isopentenyl diphosphate + NAD(P)+ + H2O 2-C-methyl-D-erithritol-4-phosphate pathway Aquifex aeolicus (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H
-
r

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.17.7.4 60
-
-
Aquifex aeolicus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.17.7.4 48 72 half-maximal activity at 48°C and 72°C Aquifex aeolicus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.17.7.4 65
-
stable Aquifex aeolicus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.17.7.4 3.7
-
(E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate recombinant enzyme, pH 7.5, 60°C Aquifex aeolicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.17.7.4 7 7.5
-
Aquifex aeolicus