Acylase의 처리 시기 및 기간이 다종 생물막 형성에 미치는 영향
Effects of Acylase Treatment Episodes on Multispecies Biofilm Development
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Quorum quenching 활성을 나타내는 acylase 효소는 생물막 형성에 중요한 영향을 미친다. 본 연구에서는 acylase 처리 조건(acylase 처리 시기 및 기간)이 다종 생물막(multispecies biofilm) 형성에 미치는 영향을 규명하였다. 서로 다른 속(genus)에 속하는 9종의 박테리아로 구성된 컨소시엄을 사용하여 acylase 처리 조건에 따른 10가지 episode 에서 다양한 acylase 농도(1, 5, 10, 20, 50 mg·l-1)에 따라5일 동안 생물막을 형성시켰다. 각 농도별로 5일 동안acylase를 처리한 조건에서 acylase의 농도가 증가함에 따라생물막 형성은 억제되었다(linear regression, Y = -0.05· x + 2.37, p < 0.05, R2 = 0.88). 모든 acylase 농도 조건에서acylase를 생물막이 형성된 후에 처리한 경우(L1−L4)에 비해서 생물막 형성 시작 단계에 처리한 경우(B1−B4) 생물막형성이 더 효과적으로 억제되었다(p < 0.05). ANOVA 결과에 따르면 acylase 10 mg·l-1 이상 농도에서 acylase 처리 기간(period)은 acylase 처리 시기(application timings, beginning or later)에 상관없이 생물막 형성에 영향을 미쳤다(p < 0.05). 각 농도별 처리 시기(L1−L4 또는 B1−B4)에서 처리 기간과생물막 형성 사이의 선형 회귀 분석 결과에 따르면 acylase 10 mg·l-1 이상 농도에서 acylase 처리 기간이 증가함에 따라생물막 형성은 억제되었다(p < 0.05, 20 mg·l-1 농도의 L1−L4 제외). 시간에 따른 생물막 형성 결과에 따르면 모든 10가지episode에서 생물막은 시간에 따라 점차 증가했으며(p < 0.05), 배양 시간과 생물막 형성 사이의 선형 회귀 분석 기울기 값은 acylase를 생물막 형성 시작 단계에 처리했을 때 더 낮게 나타났다(p < 0.05). 본 연구 결과는 생물막 형성 억제에 대한 acylase의 처리 시기 및 기간의 중요성을 시사한다.
Abstract
Acylases can have a significant effect on biofilm formation owing to their quorum quenching activity. In this study, we investigated the effects of acylase treatment episodes on multispecies biofilm development. A consortium composed of 9 species belonging to different genera was allowed to form biofilms for 5 days under various treatment episodes (different treatment periods, 1, 2, 3, or 4 days; and two application timings, beginning or later) at 1, 5, 10, 20 and 50 mg·l-1 acylase concentrations. The acylase treatment for 5 days showed that acylase concentration was negative with biofilm development (linear regression, Y = -0.05·x + 2.37, p < 0.05, R2 = 0.88). Acylase was more effective in reducing biofilm formation when it was applied in the beginning (vs. in later development stage) at all acylase concentrations (p < 0.05). ANOVA indicated that treatment period was significant on biofilm formation in both application timings at ≥ 10 mg·l-1 (p < 0.05). Linearity test results showed that all slope values between period and biofilm were negative in both timings at ≥ 10 mg·l-1 (p < 0.05, except for the later application at 20 mg·l-1). When temporal biofilm dynamics were monitored at 20 mg·l-1, biofilms gradually increased with time at all treatment episodes (p < 0.05), and slope values in linear regression between biofilm and time were lower when acylase was applied in the beginning (p < 0.05). Our findings suggest the importance of the acylase treatment period and application timing on biofilm control.
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