Mazaheri Tehrani Maryam, Erfani Mostafa, Amirmozafari Nour, Nejadsattari Taher
Department of Microbiology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Radiation Application Research School, Nuclear Science and Technology Research Institute (NSTRI), Tehran, Iran.
Iran J Pharm Res. 2019 Summer;18(3):1264-1276. doi: 10.22037/ijpr.2019.1100750.
MicrocinJ25 (MccJ25) is a small ribosomally synthesized antimicrobial peptide that is produced by family especially . The present study focuses on preparation and evaluation of antimicrobial and biological properties of a new peptide derived from MccJ25. We prepared a MccJ25-derived peptide containing 14 amino acids and a single intra-molecular disulfide bond according to solid phase synthesis strategy. The purified peptide was characterized by Liquid chromatography-mass spectrometry (LC-MS) and Fourier Transform Infrared (FTIR) spectroscopy. 96-well microdilution plate assay was exerted for determination of minimum inhibitory concentration (MIC) of peptide against different bacterial strains. Cytotoxicity of the peptide derivative on HT-29 cell line assayed using MTT test. The final peptide successfully was prepared with purity more than 99.8% as determined by analytical HPLC. The evaluation of antibacterial activity of the peptide against Gram-positive and Gram- negative bacteria revealed that the peptide was very effective against 35218 with minimum inhibitory concentration (MIC) at dose 3.9 µM. The hemolytic activity toward human erythrocytes was very minimal below 0.3%. The cell viability percentage of HT-29 cell line after 24 h of contact with the peptide was more than 83%. The high sensitivity of strain to this new peptide derived from MccJ25 and through minimal toxicity to cancerous cell, suggesting that above synthesized peptide could be considered as a bioactive compound for further investigations.
微小菌素J25(MccJ25)是一种由核糖体合成的小抗菌肽,特别是由 家族产生。本研究重点关注一种源自MccJ25的新肽的抗菌和生物学特性的制备与评估。我们根据固相合成策略制备了一种含有14个氨基酸和一个分子内二硫键的源自MccJ25的肽。通过液相色谱-质谱联用(LC-MS)和傅里叶变换红外(FTIR)光谱对纯化后的肽进行了表征。采用96孔微量稀释板法测定该肽对不同细菌菌株的最低抑菌浓度(MIC)。使用MTT试验检测该肽衍生物对HT-29细胞系的细胞毒性。经分析型高效液相色谱测定,最终成功制备的肽纯度超过99.8%。对该肽对革兰氏阳性菌和革兰氏阴性菌的抗菌活性评估表明,该肽对 35218非常有效,最低抑菌浓度(MIC)为3.9 μM。对人红细胞的溶血活性极低,低于0.3%。与该肽接触24小时后,HT-29细胞系的细胞活力百分比超过83%。 菌株对这种源自MccJ25的新肽具有高敏感性,且对癌细胞毒性极小,这表明上述合成肽可被视为一种生物活性化合物用于进一步研究。