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Biocompatibility, Antioxidant and Anti-Infective Effect of Biosurfactant Produced by Marinobacter litoralis MB15

机译:Marinobacter Litoralis MB15产生的生物活性剂的生物相容性,抗氧化剂和抗感染作用

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Objective: In this study, the biosurfactant, produced by a potent non-pathogenic bacterial strain of Marinobacter litoralis MB15 was used for anti-microbial, anti-oxidant and biocompatibility assay. Materials and Methods: Biosurfactant producing bacteria were screened by oil displacement assay and emulsification assay. The best isolate was sequenced by 16S rRNA gene analysis. The biosurfactant was produced in modified Zobel marine broth media followed by the recovery of biosurfactant by solvent extraction method. The produced biosurfactant was characterized by using thin layer chromatography, Fourier transform infrared spectroscopy and Electron spray Ionization-Mass Spectroscopy analysis. 1-diphenyl-2-picrylhydrazyl (DPPH) assay, crystal violet assay, 3-(4, 5-dimethylthiazole-2-yl)-2, 5-diphenyltetrazolium bromide) (MTT) assay were performed to determine the anti-oxidant activity, anti-microbial activity and biocompatibility of biosurfactant respectively. Results: The best biosurfactant producing isolate was found to be Marinobacter litoralis . The produced biosurfactant was characterized as rhamnolipid molecules, comprising of both mono ( m/z = 358.98 [M H] , Rha-C 12:2 ; m/z = 543.29 [M K] ,Rha–C 10 -C 10 ) and di rhamnolipid homologs ( m/z = 689.34 [M K] ,Rha-Rha-C10-C10). The product showed excellent antimicrobial properties against several pathogens including Candida albicans , Escherichia coli , Vibrio parahaemolyticus , Bacillus subtilis , S. aureus , Klebsiella pneumonia and Streptococcus pyogenes . The antioxidant activity of biosurfactant was 72.6% at 5 mg/ml. The biosurfactant showed negligible cytotoxic effect to mouse L292 fibroblastic cell line. Most importantly, after 48 h of incubation, 84.7 % of cell viability on L292 was observed at 250 μg/ml for biosurfactant. Conclusion: Therefore, this investigation shows the utility of rhamnolipids as non-cytotoxic, natural antimicrobial and antioxidant agent for various industrial and biomedical application.
机译:目的:在本研究中,通过Marinobacter Literalis MB15的有效的非致病细菌菌株产生的生物活性剂用于抗微生物,抗氧化剂和生物相容性测定。材料和方法:通过油位移测定和乳化测定筛选生物表面活性剂产生的细菌。最佳的分离物被16S rRNA基因分析测序。在改性Zobel海洋肉汤培养基中制备生物活性剂,然后通过溶剂萃取方法回收生物活性剂。通过使用薄层色谱法,傅里叶变换红外光谱和电子喷雾电离质谱分析来表征生产的生物活性剂。进行1-二苯基-2-纤维酰肼(DPPH)测定,进行晶体紫度测定,3-(4,5-二甲基唑-2-基)-2,5-二苯基四唑溴化物)(MTT)测定以确定抗氧化活性分别是生物活性剂的抗微生物活性和生物相容性。结果:发现最佳的生物表面活性剂生成孤立物是Littoralis的Marinobacter。所生产的生物活性剂的特征为耳蜗分子,其包含单声道(M / Z = 358.98 [MH],RHA-C 12:2; M / Z = 543.29 [MK],RHA-C 10 -C 10)和DI rhamnolipid同源物(m / z = 689.34 [mk],rha-rha-c10-c10)。该产品展示了针对几种病原体的优异的抗菌性质,包括念珠菌阿尔伯格,大肠杆菌,Vibrio Parahaemolyticus,枯草芽孢杆菌,黄绿骨,Klebsiella肺炎和链球菌性活塞。生物表面活性剂的抗氧化活性为5mg / ml。生物表面活性剂对小鼠L292纤维细胞系显示出可忽略的细胞毒性作用。最重要的是,在48小时孵育后,在250μg/ ml的生物活性剂中观察到L292上的84.7%的细胞活力。结论:因此,本研究表明,rhamnolipids作为各种工业和生物医学应用的非细胞毒性,天然抗微生物和抗氧化剂的效用。

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