Comparative kinetic analysis of glycerol 3-phosphate cytidylyltransferase from Enterococcus faecalis and Listeria monocytogenes.

ABSTRACT:

Background

Glycerol 3-phosphate cytidylyltransferase (GCT) is an enzyme central to the synthesis of teichoic acids, components of the cell wall in gram positive bacteria. Catalysis by GCT from Enterococcus faecalis and Listeria monocytogenes has been investigated and catalytic properties compared.

Material/methods

The genes encoding GCT were cloned from genomic DNA and recombinant proteins expressed in E. coli and purified. Enzyme assays were used to determine kinetic constants kcat and Km. Chemical crosslinking provided a means to assess quaternary structure of each GCT.

Results

Recombinant Enterococcus faecalis GCT had an apparent kcat value of 1.51 s⁻¹ and apparent Km values of 2.42 mM and 4.03 mM with respect to substrates cytidine 5'-triphosphate (CTP) and glycer

SUBMITTER: Mericl AN 

PROVIDER: S-EPMC3560613 | biostudies-literature | 2012 Nov

REPOSITORIES: biostudies-literature