A general NMR method for the assay of racemase activity with optically active or optically inactive substrates

A general NMR method for the assay of racemase activity with optically active or optically inactive substrates

A rapid and sensitive method is described for determining the activity of alanine racemase (5.1.1.1) with its substrates. Alanine racemase from Bacillus subtilis catalyzes the exchange of the α hydrogen of l and d alanine, a racemic mixture of alanine as well as both the α hydrogens of glycine with...

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Veröffentlicht in:Analytical biochemistry 1975, Vol.63 (1), p.208-213
Hauptverfasser: Babu, U.M., Johnston, R.B., McNeff, L.C.
Format: Artikel
Sprache:eng
Schlagworte:
Alanine - metabolism
Bacillus subtilis - enzymology
Deuterium
Glycine - metabolism
Isomerases - analysis
Isomerases - metabolism
Kinetics
Magnetic Resonance Spectroscopy
Mathematics
Optical Rotation
Stereoisomerism
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Zusammenfassung:A rapid and sensitive method is described for determining the activity of alanine racemase (5.1.1.1) with its substrates. Alanine racemase from Bacillus subtilis catalyzes the exchange of the α hydrogen of l and d alanine, a racemic mixture of alanine as well as both the α hydrogens of glycine with D 2O. The rate of exchange which can be followed by NMR spectroscopy is proportional to the enzyme concentration. This assay has been used to test a wide variety of possible substrates and thus determine the specificity of the enzyme.
ISSN:0003-2697
1096-0309
DOI:10.1016/0003-2697(75)90206-7