A novel capillary electrophoresis-based assay method for coenzyme B-6-dependent diaminopimelate decarboxylase from Propionibacterium acnes

作者：Huang Jui Hsin; Chen Hao Ping^*

来源：Analytical Methods, 2013, 5(5): 1279-1282.

DOI：10.1039/c2ay26169c

In bacteria, diaminopimelate decarboxylase is responsible for L-lysine biosynthesis, and may become a potential drug target for treatment of multidrug resistant bacteria. A gene encoding diaminopimelate decarboxylase was cloned from Propionibacterium acnes ATCC 6922 and was over-expressed in E. coli. Without the use of radioactive labeled compound as substrate, a capillary electrophoresis-based enzyme assay method was reported in this study. The enzyme follows Michaelis-Menten kinetics with a k(cat) of 2.2 s(-1). The apparent K-m for diaminopimelate and pyridoxal-5'-phosphate is 1.6 +/- 0.3 mM and 1.3 +/- 0.1 mu M, respectively. This assay method can be automated with an auto-sampler, which will become a cheaper alternative for large-scale screening.

• 出版日期2013