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Enhanced antibacterial metabolite production through the application of statistical methodologies by a Streptomyces nogalater NIIST A30 isolated from Western Ghats forest soil

机译:通过应用统计方法从西高止山脉森林土壤中分离出的No. Streptomyces nogalater NIIST A30增强抗菌代谢物的产生

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摘要

Streptomyces strains isolated from Nelliyampathy forest soil of Western Ghats, Kerala, India were evaluated for their antibacterial efficacy against two indicator pathogenic bacteria (Escherichia coli and Staphylococcus aureus). Among 140 strains tested, sixteen recorded potent antibacterial properties and were further screened against eleven bacterial pathogens. A strain identified as Streptomyces nogalater and designated as NIIST A30 exhibited maximum inhibition against all the test pathogens. Among the eight fermentation media tested, inorganic salts starch broth recorded the best for antibacterial production. The ethyl acetate crude extract exhibited antioxidant properties with IC50 value of 30 μg/mL and had no cytotoxicity towards L6, H9c2 and RAW 264.7 cell lines up to a concentration of 50 μg/mL. Maximum metabolite production was achieved in pH 7.0 at 35°C after 7 days incubation. The significant media components for maximum metabolite production were optimized through response surface methodology employing Plackett-Burman and Box-Behnken designs. The composition of the final optimized medium was soluble starch, 14.97g; (NH4)2SO4, 2.89g; K2HPO4, 2.07g; MgSO4.7H2O, 1g; NaCl, 1g, CaCO3, 2g; FeSO4.7H2O, 1mg; MnCl2.7H2O, 1mg; and ZnSO4.7H2O, 1mg per litre of distilled water. The optimization resulted an antibacterial activity of 28±1.5mm against S. epidermidis which was in close accordance with the predicted value of 30 mm. It is also evident from the result that an increase of 86.66% antibacterial production was recorded in optimized media. The chosen method was economical, efficient and useful for future antibacterial drug discovery from a broad spectrum metabolite producer like Streptomyces nogalater NIIST A30.
机译:从印度喀拉拉邦西高止山脉的Nelliyampathy森林土壤中分离出的链霉菌菌株对两种指示性病原菌(大肠杆菌和金黄色葡萄球菌)具有抗菌作用。在140株测试菌株中,有16株记录了有效的抗菌特性,并针对11种细菌病原体进行了进一步筛选。命名为链霉菌的菌株并命名为NIIST A30的菌株表现出对所有测试病原体的最大抑制作用。在所测试的八种发酵培养基中,无机盐淀粉肉汤的抗菌效果最佳。乙酸乙酯粗提物显示出抗氧化特性,IC50值为30μg/ mL,并且对浓度达50μg/ mL的L6,H9c2和RAW 264.7细胞系均无细胞毒性。孵育7天后,在35°C的pH 7.0下达到最大的代谢产物产量。通过采用Plackett-Burman和Box-Behnken设计的响应面方法优化了用于最大代谢物生产的重要培养基成分。最终优化培养基的组成是可溶性淀粉14.97g; (NH4)2SO4,2.89g; K2HPO4,2.07g; MgSO4.7H2O,1g;氯化钠1g,碳酸钙2g; FeSO4.7H2O,1毫克; MnCl2.7H2O,1mg;每升蒸馏水1mg和ZnSO4.7H2O。优化后,对表皮葡萄球菌的抗菌活性为28±1.5mm,与预期的30mm值非常接近。从结果还可以看出,在优化的培养基中记录了86.66%的抗菌素生成量。选择的方法经济,有效并且可用于将来从宽谱代谢产物生产商(如链霉菌NIIST A30)发现抗菌药物。

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