Expression and Characterization of Serotype 2 <b><i>Streptococcus suis </i></b>Arginine Deiminase

Abstract

Background: Arginine deiminase (ArcA) has been speculated to facilitate the intracellular survival of Streptococcus suis under acidic conditions. However, the physical and biological properties and function of SS2-ArcA have not yet been elucidated. Methods: Recombinant SS2-ArcA (rSS2-ArcA) was expressed and purified using Ni-NTA affinity chromatography. Under various pH and temperature conditions, the enzymatic properties of purified rSS2-ArcA and crude native SS2-ArcA were determined. Results: The SS2-arcA-deduced amino acid sequence contained a conserved catalytic triad (Cys399-His273-Glu218). The optimum temperature and pH of 47-kDa rSS2-ArcA and crude native SS2-ArcA were 42°C and pH 7.2. The rSS2-ArcA and crude native SS2-ArcA were stable for 3 h at 4 and 25°C, respectively. The pH stability and dependency tests suggested that rSS2-ArcA and crude native SS2-ArcA were functionally active in acidic conditions. The <smlcap>L</smlcap>-arginine substrate binding affinity (K_m) values of rSS2-ArcA (specific activity 16.00 U/mg) and crude native SS2-ArcA (specific activity 0.23 U/mg) were 0.058 and 0.157 m<smlcap>M</smlcap>, respectively. rSS2-ArcA exhibited a weak binding affinity with the common ArcA inhibitors <smlcap>L</smlcap>-canavanine and <smlcap>L</smlcap>-NIO. Furthermore, the partial inactivation of SS2-ArcA significantly impaired the viability and growth of SS2 at pH 4.0, 6.0, and 7.5. Conclusions: This study profoundly demonstrated the involvement of ArcA enzymatic activity in S. suis survival under acidic conditions.

Streptococcus suis in Brazil: Genotypic, Virulence, and Resistance Profiling of Strains Isolated from Pigs between 2001 and 2016

“…Meanwhile, the arcA gene was detected in 98.6% (212/215) of the studied strains. Arginine deiminase, encoded by the arcA gene, is described as a facilitating factor for S. suis survival under acidic conditions [26]. Previous studies have also identified a high frequency of this gene in invasive and respiratory S. suis strains isolated from swine and humans [27,28], which suggests that it is not a good virulence marker.…”

In silico and in vitro analysis of recombinant arginine deiminase from Pseudomonas furukawaii as a potential anticancer enzyme

“…After the in silico predictions, in vitro e cacy of PfADI as a potential anticancer drug candidate was validated, the arcA gene of PfADI was cloned and expressed in E. coli. Previously, ADI from various organisms viz Streptococcus sanguis, Lactococcus lactis, Enterococcus faecalis, M. arginini have been cloned and over-expressed in E. coli 3,[38][39][40] with diverse aims such as to understand its importance in cell growth, arginine metabolism or the role as anticancer candidate 41 . The arcA gene from two species of genus Pseudomonas namely P. aeruginosa and P. plecoglossicida have also been cloned in the E. coli host 31,42 .…”

“…ADI is the major enzyme of the arginine deiminase pathway in prokaryotes that serve as a non-glycolytic pathway for energy generation 2 . Besides energy production and arginine catabolism, ADI also protects the bacteria from acidic environment by generation of ammonia 3 . The enzyme is widely present in bacteria and few lower protozoa but has not been reported in mammals 4 .…”

Influence of streptococcal arginine deiminase on the leukocyte infiltration in murine air pouch model

“…ADS способствует выживанию бактерий в условиях дефицита кислорода и нутриентов, а также и при снижении рН среды в разных биологических нишах. В рамках этой системы АД катализирует гидролиз L-аргинина с образованием L-цитруллина и аммиака, поэтому активность фермента может приводить к истощению L-аргинина в микроокружении клеток организма-хозяина при инфекции [13,18,30].…”