A new robust kinetic assay for DAP epimerase activity.

Biochimie (2013-07-11)
Lilian Hor, Martin G Peverelli, Matthew A Perugini, Craig A Hutton
ABSTRACT

DAP epimerase is the penultimate enzyme in the lysine biosynthesis pathway. The most versatile assay for DAP epimerase catalytic activity employs a coupled DAP epimerase-DAP dehydrogenase enzyme system with a commercial mixture of DAP isomers as substrate. DAP dehydrogenase converts meso-DAP to THDP with concomitant reduction of NADP(+) to NADPH. We show that at high concentrations, accumulation of NADPH results in inhibition of DAPDH, resulting in spurious kinetic data. A new assay has been developed employing DAP decarboxylase that allows the reliable characterisation of DAP epimerase enzyme kinetics.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2,6-Diaminopimelic acid, ≥98% (TLC)
Sigma-Aldrich
2,6-Diaminopimelic acid, ≥97.0% (NT)

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