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L6777 Sigma-Aldrich

Lipoamide Dehydrogenase from bovine intestinal mucosa

ammonium sulfate suspension, 100-200 units/mg protein (biuret), secondary activity: 3-15 units/mg protein lipoic dehydrogenase, secondary activity: 2-10 units/mg protein “native” diaphorase (NADH)

Synonym: Diaphorase, Lipoyl Dehydrogenase, NADH:lipoamide oxidoreductase

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Properties

form   ammonium sulfate suspension
foreign activity   NADPH diaphorase, NADH oxidase, NADPH oxidase, and NADPH lipoamide dehydrogenase <1 units/mg protein
storage temp.   2-8°C

Description

Packaging

Sold on the basis of lipoamide dehydrogenase units.

Other Notes

The name "Diaphorase" has been loosely applied to several enzymes which catalyze the oxidation of either β-NADH or β-NADPH in the presence of an electron acceptor such as methylene blue or 2,6-dichlorophenol-indophenol. Many different assay procedures and "units" are used.
Diaphorases which are specific for either β-NADH or β-NADPH are known. The pig heart enzyme of Straub seems to have native diaphorase (β-NADH specific) as well as lipoic and lipoamide dehydrogenase activities. It is reported to be a single protein. However, Massey reports that "diaphorase" is probably a denatured lipoamide dehydrogenase. Pre-incubation of the pig heart preparation with Cu2+ reduces the lipoamide dehydrogenase activity and proportionately increases the β-NADH diaphorase activity. In our laboratory, we have demonstrated this copper effect to some degree on the pig heart enzyme, but no appreciable effect was observed on the Clostridium kluyveri or torula yeast preparations. The lipoamide dehydrogenase: diaphorase ratio is a measure of the denaturation.

Unit Definition

One unit will reduce 1.0 μmole of DL-lipoamide to DL-dihydrolipoamide per min at pH 6.5 at 25 °C.

Physical form

Suspension in 3.2 M (NH4)2SO4 solution, pH approx. 6.0

Safety & Documentation

Safety Information

WGK Germany 
3
Protocols & Articles

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D5540 Diaphorase from Clostridium kluyveri, lyophilized powder, 3.0-20.0 units/mg protein (biuret)

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C0400 Carmustine, ≥98%

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