Department of Microbiology and Immunology, College of Medicine, Seoul National University, Seoul, Republic of Korea.
Department of Biomedical Sciences, College of Medicine, Seoul National University, Seoul, Republic of Korea.
J Microbiol. 2023 Jan;61(1):121-129. doi: 10.1007/s12275-022-00009-1. Epub 2023 Jan 31.
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 (IC) 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 IC 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 cell-based drug screening systems using rMpg-LuxG13 strain for the discovery of novel anti-mycobacterial drugs.
我们最近的基于基因组的研究表明，分枝杆菌（Mpg）已经进化为更适应自由生活环境中的阿米巴体内的生活方式，并且其在棘阿米巴中的增强的体内存活能力也得到了证明。在这里，我们试图研究分枝杆菌（Mpg）在抗分枝杆菌药物筛选系统中的潜在用途。我们的数据表明，分枝杆菌（Mpg）比密切相关的物种分枝杆菌（Mmar）和分枝杆菌（M. gordonae）对各种抗生素更敏感，这进一步支持了其在环境中的体内生活方式，这可以解释其免受环境伤害的原因。此外，我们使用携带荧光素酶报告载体的重组分枝杆菌（rMpg-LuxG13）菌株开发了两种基于细菌全细胞的药物筛选系统：一种直接应用于 rMpg-LuxG13，另一种通过 rMpg-LuxG13 与棘阿米巴的相互作用进行药物筛选。直接应用于 rMpg-LuxG13 的结果显示，分枝杆菌（Mpg）对利福平、异烟肼、克拉霉素和环丙沙星的抑制浓度 50（IC）值比分枝杆菌（Mmar）低。通过 rMpg-LuxG13 与棘阿米巴的相互作用应用药物筛选系统也显示了分枝杆菌（Mpg）对利福平的 IC 值比分枝杆菌（Mmar）低。总之，我们的数据表明分枝杆菌（Mpg）比其他菌株对各种抗生素更敏感。此外，我们的数据还证明了使用 rMpg-LuxG13 菌株的两种基于全细胞的药物筛选系统用于发现新型抗分枝杆菌药物的可行性。
