Microbiology & Fermentation Technology Department, CSIR-Central Food Technological Research Institute, Mysore, Karnataka, India.
Bioprocess Biosyst Eng. 2024 May;47(5):683-695. doi: 10.1007/s00449-024-02998-2. Epub 2024 Mar 23.
One of the significant challenges during the purification and characterization of antimicrobial peptides (AMPs) from Bacillus sp. is the interference of unutilized peptides from complex medium components during analytical procedures. In this study, a semi-synthetic medium was devised to overcome this challenge. Using a genetic algorithm, the production medium of AMP is optimized. The parent organism, Bacillus licheniformis MCC2514, produces AMP in very small quantities. This AMP is known to inhibit RNA biosynthesis. The findings revealed that lactose, NHCl and NaNO were crucial medium constituents for enhanced AMP synthesis. The potency of the AMP produced was studied using bacterium, Kocuria rhizophila ATCC 9341. The AMP produced from the optimized medium was eightfold higher than that produced from the unoptimized medium. Furthermore, activity was increased by 1.5-fold when cultivation conditions were standardized using the optimized medium. Later, AMP was produced in a 5 L bioreactor under controlled conditions, which led to similar results as those of shake-flask production. The mode of action of optimally produced AMP was confirmed to be inhibition of RNA biosynthesis. Here, we demonstrate that improved production of AMP is possible with the developed semi-synthetic medium recipe and could help further AMP production in an industrial setup.
从芽孢杆菌属中纯化和表征抗菌肽 (AMPs) 的过程中，面临的一个重大挑战是在分析过程中，复杂培养基成分中的未利用肽会产生干扰。在本研究中，设计了一种半合成培养基来克服这一挑战。使用遗传算法对 AMP 的生产培养基进行了优化。原始生物体地衣芽孢杆菌 MCC2514 少量产生 AMP。这种 AMP 已知会抑制 RNA 生物合成。研究结果表明，乳糖、NHCl 和 NaNO 是增强 AMP 合成的关键培养基成分。使用细菌节杆菌 ATCC 9341 研究了所产生的 AMP 的效力。与未优化的培养基相比，从优化的培养基中产生的 AMP 增加了 8 倍。此外，当使用优化的培养基标准化培养条件时，活性增加了 1.5 倍。随后，在控制条件下在 5 L 生物反应器中生产 AMP，得到的结果与摇瓶生产的结果相似。优化产生的 AMP 的作用模式被确认为抑制 RNA 生物合成。在这里，我们证明使用开发的半合成培养基配方可以实现 AMP 产量的提高，这有助于在工业设备中进一步生产 AMP。
