Kinetic characterization of arginine deiminase and carbamate kinase from Streptococcus pyogenes M49

Abstract:

Streptococcus pyogenes (group A Streptococcus, GAS) is an important human pathogen causing mild superficial infections of skin and mucous membranes, but also life-threatening systemic diseases. S. pyogenes and other prokaryotic organisms use the arginine deiminase system (ADS) for survival in acidic environments. In this study, the arginine deiminase (AD), and carbamate kinase (CK) from S. pyogenes M49 strain 591 were heterologously expressed in E. coli DH5α, purified, and kinetically characterized. AD and CK from S. pyogenes M49 share high amino acid sequence similarity with the respective enzymes from Lactococcus lactis subsp. lactis IL1403 (45.6% and 53.5% identical amino acids) and Enterococcus faecalis V583 (66.8% and 66.8% identical amino acids). We found that the arginine deiminase of S. pyogenes is not allosterically regulated by the intermediates and products of the arginine degradation (E. g, ATP, citrulline, carbamoyl phosphate). The Km and Vmax values for arginine were 1.13±0.12 mM (mean ± SD) and 1.51±0.07 μmol/min/mg protein. The carbamate kinase is inhibited by ATP but unaffected by arginine and citrulline. The Km and Vmax values for ADP were 0.72±0.08 mM and 1.10±0.10 μmol/min/mg protein and the Km for carbamoyl phosphate was 0.65±0.07 mM. The optimum pH and temperature for both enzymes were 6.5 and 37°C, respectively.

SEEK ID: https://fairdomhub.org/publications/211

DOI: 10.1016/j.pep.2013.07.002

Projects: SysMO-LAB

Journal: Protein Expression and Purification

Citation:

Date Published: 1st Jul 2013

Author: Not specified

Help
help Creator
Citation
Hering, S., Sieg, A., Kreikemeyer, B., & Fiedler, T. (2013). Kinetic characterization of arginine deiminase and carbamate kinase from Streptococcus pyogenes M49. Protein Expression and Purification, 91(1), 61–68. http://doi.org/10.1016/j.pep.2013.07.002
Activity

Views: 2911

Created: 16th Jul 2013 at 16:30

Last updated: 16th Jul 2013 at 16:30

help Attributions

None

Related items

Powered by
Seek new full
(v.1.8.2)
Copyright © 2008 - 2019 The University of Manchester and HITS gGmbH