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Diaphorase 22 from Recombinant E.coli

Cat No.
NATE-1938
Description
A flavoprotein (FAD). A component of the multienzyme 2-oxo-acid dehydrogenase complexes. In the pyruvate dehydrogenase complex, it binds to the core of EC 2.3.1.12, dihydrolipoyllysine-residue acetyltransferase, and catalyses oxidation of its dihydrolipoyl groups. It plays a similar role in the oxoglutarate and 3-methyl-2-oxobutanoate dehydrogenase complexes. Another substrate is the dihydrolipoyl group in the H-protein of the glycine-cleavage system (click here for diagram), in which it acts, together with EC 1.4.4.2, glycine dehydrogenase (decarboxylating), and EC 2.1.2.10, aminomethyltransferase, to break down glycine. It can also use free dihydrolipoate, dihydrolipoamide or dihydrolipoyllysine as substrate. This enzyme was first shown to catalyse the oxidation of NADH by methylene blue; this activity was called diaphorase. The glycine cleavage system is composed of four components that only loosely associate: the P protein (EC 1.4.4.2), the T protein (EC 2.1.2.10), the L protein (EC 1.8.1.4) and the lipoyl-bearing H protein.
Source
E. coli
Form
Lyophilized
Enzyme Commission Number
EC 1.8.1.4
Activity
>150 U/mg protein
CAS No.
9001-18-7
Contaminants
(as Diaphorase activity = 100 %)
Adenylate kinase < 0.01 %
NADH oxidase < 0.20 %
Molecular Weight
ca. 110,000
pH Stability
6.0 – 9.0
Michaelis Constant
(50 mM HEPES buffer, pH 7.0, at 30 °C)
3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) 0.345 mM
NADH 0.033 mM
Unit Definition
One unit of activity is defined as the amount of Diaphorase that forms 1 μmol of NAD+ per minute at 30 °C.
Optimum pH
8
Thermal stability
No detectable decrease in activity up to 70 °C.
Storage
Store at -20°C
Synonyms
LDP-Glc; LDP-Val; dehydrolipoate dehydrogenase; diaphorase; dihydrolipoamide dehydrogenase; dihydrolipoamide:NAD+ oxidoreductase; dihydrolipoic dehydrogenase; dihydrothioctic dehydrogenase; lipoamide dehydrogenase (NADH); lipoamide oxidoreductase (NADH); lipoamide reductase; lipoamide reductase (NADH); lipoate dehydrogenase; lipoic acid dehydrogenase; lipoyl dehydrogenase; protein-6-N-(dihydrolipoyl)lysine:NAD+ oxidoreductase
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