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Literature summary for 1.5.1.30 extracted from

  • Campbell, Z.T.; Baldwin, T.O.
    Fre Is the Major Flavin Reductase Supporting Bioluminescence from Vibrio harveyi Luciferase in Escherichia coli (2009), J. Biol. Chem., 284, 8322-8328.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.5.1.30

Activating Compound

Activating Compound Comment Organism Structure
NADH
-
 Escherichia coli
NADPH
-
 Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
PCR-amplification, expression in Escherichia coli Bl21 (deltaDE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
additional information Fre oxidoreductase deletion strain shows 99% reduced bioluminescence in coupled assay with luciferase Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0009
-
 FMN coupled oxidoreductase-luciferase assay, kcat/Km = 73/(M*min), in 100 mM Na+/K+ phosphate buffer with 100 mM NaCl, pH 7.0, 1 microM Fre oxidoreductase, 10 microM decanal, 10 microM NADPH, 5 microM luciferase Escherichia coli
0.0013
-
 riboflavin coupled oxidoreductase-luciferase assay, kcat/Km = 1257/(M*min), in 100 mM Na+/K+ phosphate buffer with 100 mM NaCl, pH 7.0, 1 microM Fre oxidoreductase, 10 microM decanal, 10 microM NADPH, 5 microM luciferase, riboflavin is favoured by Fre oxidoreductase but a poor substrate for bacterial luciferase Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
 SDS-PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
FMN + NADH + H+ Escherichia coli
-
 FMNH2 + NAD+
-
 ?
FMN + NADPH Escherichia coli
-
 FMNH2 + NADP+ + H+
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
crude lysate assay with briefly centrifuged cells, resuspended in B-PER bacterial protein extraction reagent, recombinant pZCFRE1 expression product is clarified by centrifugation and applied to a custom nickel affinity column, dialyzed into buffer containing 100 mM Na+/K+ phosphate and 100 mM NaCl, pH 7.0 Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
FMN + NADH + H+
-
 Escherichia coli FMNH2 + NAD+
-
 ?
FMN + NADPH
-
 Escherichia coli FMNH2 + NADP+ + H+
-
 ?
FMN + NADPH + H+
-
 Escherichia coli FMNH2 + NADP+
-
 ?
riboflavin + NADPH + H+
-
 Escherichia coli reduced riboflavin + NADP+
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 26000 Escherichia coli

Synonyms

Synonyms Comment Organism
flavin oxidoreductase
-
 Escherichia coli
flavin reductase
-
 Escherichia coli
Fre oxidoreductase
-
 Escherichia coli

General Information

General Information Comment Organism
physiological function provides reducing equivalents required to drive the luciferase genes in Escherichia coli but is not coupled to luciferase Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0012
-
 FMN coupled oxidoreductase-luciferase assay, in 100 mM Na+/K+ phosphate buffer with 100 mM NaCl, pH 7.0, 1 microM Fre oxidoreductase, 10 microM decanal, 10 microM NADPH, 5 microM luciferase Escherichia coli
0.021
-
 riboflavin coupled oxidoreductase-luciferase assay, in 100 mM Na+/K+ phosphate buffer with 100 mM NaCl, pH 7.0, 1 microM Fre oxidoreductase, 10 microM decanal, 10 microM NADPH, 5 microM luciferase, riboflavin is favoured by Fre oxidoreductase but a poor substrate for bacterial luciferase Escherichia coli