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Literature summary for 1.17.7.4 extracted from

  • Wolff, M.; Seemann, M.; Tse Sum Bui, B.; Frapart, Y.; Tritsch, D.; Estrabot, A.G.; Rodriguez-Concepcion, M.; Boronat, A.; Marquet, A.; Rohmer, M.
    Isoprenoid biosynthesis via the methylerythritol phosphate pathway: the (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (LytB/IspH) from Escherichia coli is a [4Fe-4S] protein (2003), FEBS Lett., 541, 115-120.
    View publication on PubMed

Inhibitors

Inhibitors Comment Organism Structure
Co2+ weak inhibition of the enzyme, when FAD is used as reducing cofactor Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ Co2+ and Mn2+, at 0.5 mM, lower the content of isopentenyl diphosphate in the mixture of both products, isopentenyl diphosphate and dimethylallyl diphosphate Escherichia coli
Fe2+ enzyme possesses a dioxygen-sensitive [4Fe-4S] cluster Escherichia coli
Mn2+ activates Escherichia coli
Mn2+ Co2+ and Mn2+, at 0.5 mM, lower the content of isopentenyl diphosphate in the mixture of both products, isopentenyl diphosphate and dimethylallyl diphosphate Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36591
-
2 * 36591, recombinant His-tagged enzyme, amino acid sequence determination Escherichia coli
72000
-
recombinant His-tagged enzyme, gel filtration Escherichia coli

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
gene ispH, enzyme LytB
-

Oxidation Stability

Oxidation Stability Organism
enzyme possesses a O2-sensitive [4Fe-4S] cluster Escherichia coli

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged apoprotein from Escherichia coli strain M15, to homogeneity Escherichia coli

Renatured (Commentary)

Renatured (Comment) Organism
reconstitution of purified apoenzyme with prosthetic group [Fe4-S4] cluster Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.002
-
purified apoenzyme, with FAD and NADPH as reducing cofactors Escherichia coli
0.011
-
purified apoenzyme, with photoreduced deazaflavin as reducing cofactor Escherichia coli
0.03
-
reconstituted purified enzyme, with photoreduced deazaflavin as reducing cofactor Escherichia coli
0.044
-
reconstituted purified enzyme, with FAD and NADPH as reducing cofactors Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+
-
Escherichia coli isopentenyl diphosphate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O Co2+ and Mn2+ lower the content of isopentenyl diphosphate in the mixture of both products, isopentenyl diphosphate and dimethylallyl diphosphate r
(E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+
-
Escherichia coli dimethylallyl diphosphate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
r
isopentenyl diphosphate + NAD(P)+ + H2O part of isoprenoid biosynthesis Escherichia coli (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H
-
r
isopentenyl diphosphate + NAD(P)+ + H2O 2-C-methyl-D-erithritol-4-phosphate pathway Escherichia coli (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H
-
r

Subunits

Subunits Comment Organism
dimer 2 * 36591, recombinant His-tagged enzyme, amino acid sequence determination Escherichia coli

Cofactor

Cofactor Comment Organism Structure
deazaflavin requires a reducing system, e.g. flavodoxin/flavodoxin reductase or NADPH or the photoreduced deazaflavin radical Escherichia coli
FAD requires a reducing system, e.g. flavodoxin/flavodoxin reductase or NADPH or the photoreduced deazaflavin radical Escherichia coli
additional information enzyme possesses a dioxygen-sensitive [4Fe-4S] cluster as prosthetic group Escherichia coli
NADP+
-
Escherichia coli
NADPH requires a reducing system, e.g. flavodoxin/flavodoxin reductase or NADPH or the photoreduced deazaflavin radical Escherichia coli