[논문]산성 pH 조건에서 차아염소산나트륨의 항균 활성 향상

손현빈; 배원빈; 지광환

doi:10.48022/mbl.2204.04014

산성 pH 조건에서 차아염소산나트륨의 항균 활성 향상
Enhanced Antibacterial Activity of Sodium Hypochlorite under Acidic pH Condition 원문보기

Microbiology and biotechnology letters = 한국미생물·생명공학회지, v.50 no.2, 2022년, pp.211 - 217

손현빈 (금오공과대학교응용화학과) , 배원빈 (금오공과대학교응용화학과) , 지광환 (금오공과대학교응용화학과)

초록
AI-Helper

차아염소산나트륨(NaClO)은 병원 및 식품산업 분야에서 널리 사용되는 소독제로 세균, 곰팡이, 바이러스에 대해서도 항균 활성이 있다. 차아염소산나트륨의 항균 활성은 용액의 pH에 의해 조절되는 안정적인 HClO 농도의 유지에 있다. 차아염소산(HClO)은 화학적으로 중성이므로 세균의 막에 쉽게 침투할 수 있으며 차아염소산나트륨의 항균 활성은 차아염소산염 이온(ClO^-)보다는 용액 내 HClO 농도에 의존하리라 사료된다. 본 연구에서 pH 조절에 따른 차아염소산나트륨의 항균 활성을 time kill test와 차아염소산나트륨 처리 전후의 활성산소종(ROS) 및 ATP 농도 변화로 조사하였다. 또한 전계방출형 주사 전자 현미경(FE-SEM)을 통하여 세포벽의 파괴정도를 확인하였다. pH 5 조건에서 5 ppm 차아염소산나트륨은 Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) 균에 대하여 99.9%의 항균 활성을 나타내었고, ROS 생성량은 pH 7 조건보다 48% 증가하였다. 또한, pH 5 조건의 차아염소산나트륨에 노출된 E. coli와 S. aureus의 ATP 농도가 각각 94%와 91% 감소하였다. FE-SEM 결과, pH 5 조건에 노출된 균의 세포벽이 파괴된 것을 확인하였다. 본 연구결과를 종합해보면, pH를 조절하는 것 만으로 5 ppm 농도의 차아염소산나트륨의 항균 활성을 향상시킬 수 있음을 시사한다.

Abstract ▼ AI-Helper

Sodium hypochlorite (NaClO) is a disinfectant widely used in hospitals and food industries because of its antimicrobial activity against not only bacteria but also fungi and virus. The antibacterial activity of NaClO lies in the maintenance of a stable hypochlorous acid (HClO) concentration, which is regulated by pH of the solution. HClO can easily penetrate bacterial cell membrane due to its chemical neutrality and the antibacterial activity of NaClO is thought to depend on the concentration of HClO in solution rather than hypochlorite ions (ClO-). In this study, we investigated the antibacterial activity of NaClO according to pH adjustment by means of time kill test and assays of Reactive Oxygen Species (ROS) and adenosine triphosphate (ATP) concentration changes before and after NaClO treatment. We also investigated that the degree of cell wall destruction through field emission scanning electron microscopy (FE-SEM). Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) exposed to 5 ppm NaClO at pH 5 exhibited 99.9% mortality. ROS production at pH 5 was 48% higher than that produced at pH 7. In addition, the ATP concentration in E. coli and S. aureus exposed to pH 5 decreased by 94% and 91%, respectively. As a result of FE-SEM, it was confirmed that the cell wall was destroyed in the bacteria by exposing to pH 5 NaClO. Taken together, our results indicate that the antibacterial activity of 5 ppm NaClO can be improved simply by adjusting the pH.

주제어

표/그림 (5)

그림 Fig. 1. Time kill test data of NaClO against E. coli (A~D) and S. aureus (E~H). Colonies were counted in following: (A, E) pH 7 controls (without NaClO treatment), (B, F) pH 5 controls (without NaClO treatment), (C, G) pH 7 samples (with NaClO treatment for 30 min), (D, H) pH 5 samples (with NaClO treatment for 30 min).
표 Table 1. Colony forming unit by time kill test after 5 ppm NaClO treatment against two strains.
그림 Fig. 2. ROS generation in bacterial cells treated with NaClO. Bacteria were pretreated for 30 min at 37℃ with 5 μM 2,7- dichlorofluorescein diacetate (DCFH-DA). And then each strain was incubated with 5 ppm NaClO for 30 min between pH 3 to 9. The fluorescence intensity was measured at an excitation/ emission wavelength of 488/535 nm.
그림 Fig. 3. pH effects for ATP generation of bacteria after 30 min treatment with 5 ppm NaClO. Bioluminescence of intracellular ATP was evaluated by the luciferin-luciferase reagent and measured by microplate reader with advanced detection capabilities.
그림 Fig. 4. Morphological changes of E. coli (A~D) and S. aureus (E~H) were observed by field emission scanning electron microscopy after treatment with 5 ppm NaClO. (A, E) pH 7 controls (without NaClO treatment), (B, F) pH 5 controls (without NaClO treatment), (C, G) pH 7 samples (with NaClO treatment for 30 min), (D, H) pH 5 samples (with NaClO treatment for 30 min). Red arrows exhibit the cell wall pores.

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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