A high-performance liquid chromatography method for the simultaneous assay of diaminopimelate epimerase and decarboxylase.
A sensitive and comparatively simple method for the assay of diaminopimelate (DAP) decarboxylase, which simultaneously monitors DAP epimerase activity, in the reverse of the biosynthetic direction, is described. The substrate, meso-DAP and products LL-DAP and L-lysine are derivatized with o-phthaldialdehyde and resolved by reversed-phase high-performance liquid chromatography. Separation is achieved on a Spherisorb C18 column using a gradient elution system. This technique offers a high degree of sensitivity as the detection method described can measure picomole quantities of substrate and products.
Weir
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AN
Institute for Biotechnological Studies, University of Kent, Canterbury, United Kingdom.
Bucke
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C
Holt
G
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Lilly
M D
MD
Bull
A T
AT
D000591:Amino Acid Isomerases; D001412:Bacillus subtilis / Q000201:enzymology; D001426:Bacterial Proteins; D002262:Carboxy-Lyases / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007535:Isomerases / Q000032:analysis*; D007536:Isomerism; D008239:Lysine / Q000032:analysis; D019998:Racemases and Epimerases / Q000032:analysis*
D001426:Bacterial Proteins; D002262:Carboxy-Lyases; C079912:LysA protein, Bacteria; C020031:diaminopimelic acid decarboxylase; D007535:Isomerases; D019998:Racemases and Epimerases; D000591:Amino Acid Isomerases; C042032:diaminopimelate epimerase; D008239:Lysine