Pournejati Roya, Karbalaei-Heidari Hamid Reza
Molecular Biotechnology Laboratory, Department of Biology, Faculty of Science, Shiraz University, Shiraz, Iran.
Avicenna J Med Biotechnol. 2020 Apr-Jun;12(2):116-123.
Isolation, introduction and producing bioactive compounds from bacteria, especially marine bacteria, is an attractive research area. One of the main challenges of using these metabolites as drug and their industrialization is the optimization of production conditions.
In the present study, the response surface methodology was applied to optimize the production of a cytotoxic extract (C-137-R) by () strain RP137. Initially, among the three carbon and three nitrogen sources, rice starch and potassium nitrate were selected as the best, with cell toxicity equal to IC50=54.4 and 45.1 in human lung and liver cancer cell lines, respectively (A549 and HepG2). In the next step, fractional factorial design was performed to survey effect of seven physical and chemical factors on the amount of production, and the most important factors including carbon and nitrogen sources with the positive effect and the sea salt with negative effect were determined. Finally, using the central composite design with 20 experiments, the best concentrations of rice starch and potassium nitrate (1.5%) and sea salt (1%) were obtained.
The average amount of dried extract produced in the optimum conditions was 131.1 and the best response was 71.45%, which is more than 28-fold better than the pre-optimized conditions.
In general, it can be suggested that the use of modern statistical methods to optimize environmental conditions affecting the growth and metabolism of bacteria can be a highly valuable tool in industrializing the production of bioactive compounds.
从细菌尤其是海洋细菌中分离、引入并生产生物活性化合物是一个具有吸引力的研究领域。将这些代谢产物用作药物及其产业化的主要挑战之一是生产条件的优化。
在本研究中，应用响应面法优化()菌株RP137产生细胞毒性提取物（C - 137 - R）的条件。最初，在三种碳源和三种氮源中，选择大米淀粉和硝酸钾作为最佳碳源和氮源，其对人肺癌和肝癌细胞系（A549和HepG2）的细胞毒性分别等于IC50 = 54.4和45.1。下一步，进行析因设计以研究七个物理和化学因素对产量的影响，并确定了具有积极影响的碳源和氮源以及具有负面影响的海盐等最重要因素。最后，通过20次实验的中心复合设计，获得了大米淀粉、硝酸钾（1.5%）和海盐（1%）的最佳浓度。
在最佳条件下产生的干燥提取物平均量为131.1，最佳响应为71.45%，比优化前的条件提高了28倍多。
总体而言，可以认为使用现代统计方法优化影响细菌生长和代谢的环境条件可能是生物活性化合物生产工业化中非常有价值的工具。
