Dr Larissa Balakireva, CEO & Founder of NovoCIB, was awarded with the Trophy of
"Femmes en Or 2011, Femme de l'Innovation"
in September 2011

Ref. #E-Nov8
Quantity Price*
25 Units € 275.00
50 Units € 480.00
100 Units € 825.00

Bacterial Recombinant FMN Reductase (FRE)

Provided in stable lyophilized form and shipped without dry ice.
* Pricing updated December 12th, 2013.

Bulk quantity available. Click to order or to ask for Quotation

Synonyms: NAD(P)H:flavin oxidoreductase, NAD(P)H:flavin mono-nucleotide oxidoreductase, NAD(P)H(2):FMN oxidoreductase, NAD(P)H-FMN reductase, NAD(P)H-dependent FMN reductase, NAD(P)H:FMN oxidoreductase, riboflavin mononucleotide reductase, flavin mononucleotide reductase

NOVOCIB's bacterial (E. coli) NAD(P)H-dependent FMN-oxidoreductase is a recombinant protein of ca. 26kDa overexpressed in E.coli. The sequence of cloned Fre (SwissProt accession number P0AEN1) was confirmed by DNA sequencing (100% identity). NAD(P)H:flavin oxidoreductases (or flavin reductases) catalyze the reduction of riboflavin, FMN, and FAD by NADPH and NADH. These enzymes are present in all microorganisms, including luminous marine bacteria, and also in mammals. Flavin reductases have been classified into two groups. The first group includes flavoproteins that use a flavin prosthetic group for the electron transfer from NAD(P)H to the flavin substrate (flavin reductase P from Vibrio harveyi). In the second group, enzymes do not contain any prosthetic group. Fre, NAD(P)H:flavin oxidoreductase from E. coli, belongs to this second group of enzymes.

Applications Coupling of bacterial luciferase to FMN-NAD(P)H oxidoreductase has been used to provide ultrasensitive analytical tools for the quantification of NADH and the substrates of NADH-, NADPH- dependent enzymes (e.g. glucose, lactate, malate, ethanol, sorbitol, oxaloacetate). Although FMN-reductase often present in luciferase enzyme preparations may be sufficient for producing light in the presence of NAD(P)H, highly purified and characterized Fre enzyme can offer some advantages such as an increased sensitivity, better control of the signal intensity and duration, and saving of the luciferase enzyme (see below, Figure 1).

Unit Definition: One unit of FMN-reductase converts 1.0 µmole of FMN and NADH to FMNH2 and NAD per minute at pH 7.9 at 37�C.

Assay conditions: 100mM Tris-HCl, pH 7.9, 250µM NADH, 25µM FMN.

Specific Activity: ≥ 2 unit/mg protein.

Purity: controlled by 10% AA SDS-PAGE.
Figure 1: Calibration curves (log-log plot) for NADH obtained using purified luciferase (50�g/ml, produced and purified by NovoCIB) in the presence (Red) or absence (Blue) of exogenous FRE (10mU/ml).

Luminescence signal (5 seconds) was measured immediately after NADH addition (5�L) to 200�L-well containing 0.1M KH2PO4 pH=6.9, 0.02% decanal, 50�M FMN, 2mg/ml BSA. Reaction was followed in a FluoroSkan Ascent FL (ThermoLabsystems) microtiter plate reader.

References (with links to PubMed)
F. Fieschi et al. (1995): The mechanism and substrate specificity of the NADPH:flavin oxidoreductase from Escherichia coli J. Biol. Chem. 270(51), 30392-400

Download this document "NovoCIB's FMN Reductase" 

Related Link
Highly Pure Bacterial Luciferase from Photobacterium phosphoreum

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