Streptomyces: isolation, optimization of culture conditions and extraction of secondary metabolites

A. Khattab, Eltahir H. Babiker, H. A. S. Supervisor
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引用次数: 48

Abstract

The objectives of this study were to isolate and identify Streptomyces from soil sediments as well as to optimize cultural growth conditions for maximum antibacterial productivity. A total of fifty soil sediments were collected from Red Sea, Sudan. The soil sediments were pretreated and cultivated on agar medium. Promising Streptomyces spp. were isolated by agar overlay method using indicator organisms. Optimization of chemical and physical culture conditions was carried out. The later was judged by assessment of antibacterial activity. Ethyl acetate was used to extract the secondary metabolite compounds. The separation of the active ingredients was performed using both thin layer chromatography (TLC) and gas chromatography-mass spectrometer (GC-MS). The results revealed nine strains of Streptomyces . Of them two (PS1 and PS28) isolates exhibited high activity against pathogenic bacteria. The optimum growth conditions were pH 7.5, temperature at 30°C, soyabean concentration 2.5 g/l, incubation period in 7 days, MgSO 4 .7H 2 O conc. 1g/l and K 2 HPO 4  conc. 2.5g/l. TLC test showed three and two fragments from metabolites of PS1 and PS28 respectively, while the GC-MS analysis revealed eight and eleven compounds with antibacterial activity of PS1 and PS28 respectively. It is concluded that marine is promising source of secondary metabolites. Khattab et al., International Current Pharmaceutical Journal, February 2016, 5(3): 27-32
链霉菌的分离、培养条件优化及次生代谢产物的提取
本研究的目的是从土壤沉积物中分离和鉴定链霉菌,并优化培养生长条件以获得最大的抗菌效率。在苏丹红海共收集了50个土壤沉积物。对土壤沉积物进行预处理,并在琼脂培养基上培养。利用指示剂琼脂覆盖法对链霉菌进行了分离。对化学和物理培养条件进行了优化。后者通过抗菌活性评价来判断。用乙酸乙酯提取次生代谢产物。采用薄层色谱(TLC)和气相色谱-质谱联用(GC-MS)对有效成分进行分离。结果发现9株链霉菌。其中2株PS1和PS28对病原菌具有较高的抑菌活性。最适生长条件为pH 7.5、温度30℃、大豆浓度2.5 g/l、培养7 d、mgso4.7 h 2 O浓度。1g/l和k2hpo4conc。2.5 g / l。薄层色谱分析显示PS1和PS28代谢产物分别有3个和2个片段,GC-MS分析显示PS1和PS28分别有8个和11个化合物具有抗菌活性。因此,海洋是次生代谢物的良好来源。Khattab等,国际药学杂志,2016,5(3):27-32
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