A new robust kinetic assay for DAP epimerase activity.
Biochimie
; 95(10): 1949-53, 2013 Oct.
Article
in En
| MEDLINE
| ID: mdl-23838343
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Corynebacterium glutamicum
/
Escherichia coli
/
Enzyme Assays
/
Amino Acid Isomerases
Language:
En
Journal:
Biochimie
Year:
2013
Document type:
Article
Affiliation country:
Australia