Djinni Ibtissem, Defant Andrea, Djoudi Warda, Chaabane Chaouch Faouzia, Souagui Samiha, Kecha Mouloud, Mancini Ines
Laboratoire de Microbiologie Appliquée, Faculté des Sciences de la Nature et de la Vie, Département de Microbiologie, Université de Bejaia, 06000, Algeria.
Bioorganic Chemistry Laboratory, Department of Physics, University of Trento, via Sommarive 14, Povo, I-38123, Trento, Italy.
Heliyon. 2019 May 27;5(5):e01695. doi: 10.1016/j.heliyon.2019.e01695. eCollection 2019 May.
The novel bioactive actinobacterial strain GSBNT10 obtained from a Saharan soil, was taxonomically characterized using a polyphasic approach. 16S rRNA gene sequence analysis supported the classification of the isolate within the genus indicating it as a novel species. The major metabolite responsible of the bioactivity was purified and structurally characterized as actinomycin D (act-D) by mass spectrometric and nuclear magnetic resonance analyses Plackett-Burman design (PBD) and response surface methodology (RSM) were applied in order to optimize the medium formulation for the production of this bioactive metabolite. By PBD experiments, NaNO, KHPO and initial pH value were selected as significant variables affecting the metabolite production. Central Composite Design (CCD) showed that adjustment of the fermentative medium at pH 8.25, KHPO at 0.2 gL and NaNO at 3.76 gL were the values suiting the production of act-D. Moreover, the results obtained by the statistical approach were confirmed by act-D detection using the HPLC equipped with a diode array detector and coupled online with electrospray-mass spectrometry (ESIMS) technique. act-D production was highly stimulated, obtaining a good yield (656.46 mgL) which corresponds to a 58.56% increase compared with the non-optimized conditions and data from LC-ESIMS technique efficiently confirmed the forecast from RSM.
从撒哈拉沙漠土壤中分离得到的新型生物活性放线菌菌株GSBNT10,采用多相分类法进行了分类鉴定。16S rRNA基因序列分析支持将该分离株归类于该属,表明它是一个新物种。通过质谱和核磁共振分析,对具有生物活性的主要代谢产物进行了纯化,并将其结构鉴定为放线菌素D(act-D)。应用Plackett-Burman设计(PBD)和响应面方法(RSM)来优化用于生产这种生物活性代谢产物的培养基配方。通过PBD实验,选择NaNO₃、KH₂PO₄和初始pH值作为影响代谢产物产量的显著变量。中心复合设计(CCD)表明,将发酵培养基的pH值调节至8.25、KH₂PO₄调节至0.2 g/L和NaNO₃调节至3.76 g/L是适合act-D生产的值。此外,通过配备二极管阵列检测器并与电喷雾质谱(ESIMS)技术在线联用的HPLC检测act-D,证实了统计方法获得的结果。act-D的产量受到极大刺激,获得了良好的产量(656.46 mg/L),与未优化条件相比增加了58.56%,并且LC-ESIMS技术的数据有效地证实了RSM的预测。