Our recent genome-based study indicated that Mycobacterium paragordonae (Mpg) has evolved to become more adapted to
an intracellular lifestyle within free-living environmental amoeba and its enhanced intracellular survival within Acanthamoeba
castellanii was also proved. Here, we sought to investigate potential use of Mpg for antimycobacterial drug screening
systems. Our data showed that Mpg is more susceptible to various antibiotics compared to the close species M. marinum
(Mmar) and M. gordonae, further supporting its intracellular lifestyle in environments, which would explain its protection
from environmental insults. In addition, we developed two bacterial whole-cell-based drug screening systems using a
recombinant Mpg stain harboring a luciferase reporter vector (rMpg-LuxG13): one for direct application to rMpg-LuxG13
and the other for drug screening via the interaction of rMpg-LuxG13 with A. castellanii. Direct application to rMpg-LuxG13
showed lower inhibitory concentration 50 (
IC50) values of rifampin, isoniazid, clarithromycin, and ciprofloxacin against
Mpg compared to Mmar. Application of drug screening system via the interaction of rMpg-LuxG13 with A. castellanii also
exhibited lower IC50
values for rifampin against Mpg compared to Mmar. In conclusion, our data indicate that Mpg is more
susceptible to various antibiotics than other strains. In addition, our data also demonstrate the feasibility of two whole cellbased
drug screening systems using rMpg-LuxG13 strain for the discovery of novel anti-mycobacterial drugs.
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