[논문]Tuberostemonine에 의한 Staphylococcus aureus의 생물막 억제 효과

염수진; 김승민; 권준혁; 정희곤

doi:10.9721/kjfst.2022.54.2.241

Tuberostemonine에 의한 Staphylococcus aureus의 생물막 억제 효과
Inhibitory effects of tuberostemonine on Staphylococcus aureus biofilm 원문보기

한국식품과학회지 = Korean journal of food science and technology, v.54 no.2, 2022년, pp.241 - 246

염수진 (충남대학교 농업생명과학대학 식품공학과) , 김승민 (한국방송통신대학교 자연과학대학 생활과학부) , 권준혁 (충남대학교 농업생명과학대학 식품공학과) , 정희곤 (충남대학교 농업생명과학대학 식품공학과)

초록
AI-Helper

본 연구는 S. aureus에 대한 tuberostemonine의 항균 및 항생물막 효과에 대하여 확인하였다. S. aureus에 대한 tuberostemonine의 생장 저해 효과가 없음을 확인하였으나, crystal violet 염색법과 CLSM 이미지 측정을 통해 tuberostemonine이 유의한 S. aureus 항생물막 효과 가지는 것을 알 수 있었다. S. aureus의 생물막 형성과 분해 관련 유전자인 icaA와 agrA의 발현은 tuberostemonine를 처리하였을 때 유의미하게 각각 감소 또는 증가하는 것으로 나타났다. 따라서 본 연구에서 생물막 형성 저해 및 분해 효과가 확인된 천연화합물인 tuberostemonine은 S. aureus의 내성 발생 위험이 적은 새로운 항생물막제제로서 사용가능 할 것으로 사료된다.

Abstract ▼ AI-Helper

Antibiotic resistance is a serious problem to food safety as well as human healthcare. To avoid this, there are several approaches for a new class of antibiotic agents that target only production of virulence factors such as biofilm without bacterial growth defect. The objective of this study was to investigate the antibiofilm activity of tuberostemonine in Staphylococcus aureus. Tuberostemonine significantly reduced the biofilm formation (26.07-47.02%) in the crystal violet assay whereas there were no effect on S. aureus growth. The dispersion in preformed biofilm was also observed by confocal laser scanning microscopy (CLSM). Quantification real-time PCR revealed that the icaA and agrA expression having an important role in biofilm production of S. aureus were strongly affected with tuberostemonine. These results suggest that tuberostemonine has potential for controlling biofilm formation and dispersion by effect on virulence regulation of S. aureus.

주제어

표/그림 (5)

그림 Fig. 1. Effect of tuberostemonine on S. aureus growth. S. aureus was incubated at 37℃ for 16 h and subcultured in 20 mL of LB broth media until its optical density (O.D._{600 nm}) reached 0.8. Then, growth of the S. aureus group and the S. aureus added with from DMSO (control) and tuberostemonine (50-200 μM) were identified by measuring their optical density every 2 h until 8 h after tuberostemonine treatment (n.s; not significant).
그림 Fig. 2. Inhibitory activities of tuberostemonine against S. aureus biofilm formation. S. aureus cells were incubated with DMSO or tuberostemonine (5-200 μM) for 16 h. The formed-biofilms were stained with 1.5% crystal violet. Measured values represent the means for triplicate wells and error bars indicated range. *p<0.05, **p<0.01.
그림 Fig. 3. Effect of tuberostemonine on biofilms pre-formed by S. aureus. Biofilms were formed at 37℃ for 24 h, and these were treated with DMSO or tuberostemonine (200 μM) for 3 h. The biofilms were stained with 1.5% crystal violet. Measured values represent the means for triplicate wells and error bars indicated range. ***p<0.001.
그림 Fig. 4. Confocal laser scanning microscopy images of S. aureus biofilms after tuberostemonine treatment. Biofilm of S. aureus were treated with tuberostemonine at 37℃ for 3 h. Confocal images of biofilms were analyzed using LSM to quantify biomass after DMSO or tuberostemonine treatment (0.1-200 μM). (A) Confocal microscopy images of Live/Dead staining of biofilms dispersed by tuberostemonine. (B) Biovolume determined from biofilm structures as depicted in Fig. 4A. *p<0.05, ***p<0.001.
그림 Fig. 5. Effect of tuberostemonine on the relative expression levels of icaA and agrA in S. aureus. The values were mean±standard deviation of three repeated processes. **p<0.01, ***p<0.001.

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