title: A high-performance liquid chromatography method for the simultaneous assay of diaminopimelate epimerase and decarboxylase.
abstract: 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.
authors.affiliation: Institute for Biotechnological Studies, University of Kent, Canterbury, United Kingdom.
authors.forename: A N
authors.initials: AN
authors.lastname: Weir
authors.forename: C
authors.initials: C
authors.lastname: Bucke
authors.forename: G
authors.initials: G
authors.lastname: Holt
authors.forename: M D
authors.initials: MD
authors.lastname: Lilly
authors.forename: A T
authors.initials: AT
authors.lastname: Bull
mesh_terms: 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*
chemical_list: 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