Panahi Chegini Parvaneh, Nikokar Iraj, Tabarzad Maryam, Faezi Sobhan, Mahboubi Arash
Department of Medicinal Biotechnology, School of Paramedicine, Guilan University of Medical Sciences, Rasht, Iran.
Medical Biotechnology Research Center, School of Paramedicine, Guilan University of Medical Sciences, Rasht, Iran.
Iran J Pharm Res. 2019 Fall;18(Suppl1):157-168. doi: 10.22037/ijpr.2019.112397.13734.
Recently, antimicrobial peptides have been introduced as potent antibiotics with a wide range of antimicrobial activities. They have also exhibited other biological activities, including anti-inflammatory, growth stimulating, and anti-cancer activities. In this study, an analog of Magainin II was designed and produced as a recombinant fusion protein. The designed sequence contained 24 amino acid residues (P24), in which Lys, His, Ser residues were substituted with Arg and also, hydrophobic Phe was replaced with Trp. Recombinant production of P24 in BL21 using pTYB21, containing chitin binding domain and intein sequence at the N-terminus of the peptide gene, resulted in 1 μg mL product from culture. Chitin column chromatography, followed by online peptide cleavage with thiol reducing agent was applied to purify the peptide. Antimicrobial activity was evaluated using five bacteria strains including and . Designed AMP exhibited promising antimicrobial activities with low minimum inhibitory concentration, in the range of 64-256 µg/mL. P24 showed potent antimicrobial activity preferably against Gram-positive bacteria, and more potent than pexiganan as a successful Magainin II analog for topical infections. In general, further modification can be applied to improve its therapeutic index.
最近，抗菌肽已作为具有广泛抗菌活性的强效抗生素被引入。它们还表现出其他生物活性，包括抗炎、生长刺激和抗癌活性。在本研究中，设计并制备了一种麦加宁II类似物作为重组融合蛋白。设计的序列包含24个氨基酸残基（P24），其中赖氨酸、组氨酸、丝氨酸残基被精氨酸取代，并且疏水性苯丙氨酸被色氨酸取代。使用pTYB21在BL21中重组生产P24，该载体在肽基因的N端包含几丁质结合结构域和内含肽序列，培养物产生了1μg/mL的产物。采用几丁质柱色谱法，随后用硫醇还原剂进行在线肽切割以纯化该肽。使用包括……和……在内的五种细菌菌株评估抗菌活性。设计的抗菌肽表现出有前景的抗菌活性，最低抑菌浓度较低，范围为64 - 256μg/mL。P24显示出强效的抗菌活性，尤其对革兰氏阳性菌有效，并且比作为用于局部感染的成功的麦加宁II类似物的pexiganan更有效。一般而言，可以进行进一步修饰以提高其治疗指数。
