Xing Li-Wei, Tian Shi-Xun, Gao Wei, Yang Na, Qu Pei, Liu Di, Jiao Jian, Wang Jue, Feng Xing-Jun
Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
Exp Ther Med. 2016 Oct;12(4):2324-2330. doi: 10.3892/etm.2016.3578. Epub 2016 Aug 5.
Fowlicidins are a group of cathelicidin antimicrobial peptides that were initially identified in chickens. Fowlicidin-2, which is composed of 31 amino acids, is widely expressed in the majority of tissues in chickens and has an important role in innate immunity. In the present study, a recombinant expression system for fowlicidin-2 was successfully constructed using X-33 and the expression vector pPICZα-A. Under the optimized fermentation conditions, 85.6 mg fowlicidin-2 with >95% purity was obtained from 1 liter culture medium following purification by ion exchange chromatography and reversed phase high performance liquid chromatography. The recombinant fowlicidin-2 exhibited broad spectrum antimicrobial activity and had a minimum inhibitory concentration ranging from 1 to 4 µM. Furthermore, recombinant fowlicidin-2 exhibited hemolytic activity, promoting 50% human erythrocyte hemolysis in the concentration range of 128-256 µM, and anticancer activity, resulting in the death of 50% of A375 human malignant melanoma cells in the concentration range of 2-4 µM. The results of the present study suggest that recombinant fowlicidin-2 may be a promising candidate for therapeutic applications.
禽杀菌肽是最初在鸡中发现的一组组织蛋白酶抗菌肽。由31个氨基酸组成的禽杀菌肽-2在鸡的大多数组织中广泛表达，在先天免疫中起重要作用。在本研究中，使用X-33和表达载体pPICZα-A成功构建了禽杀菌肽-2的重组表达系统。在优化的发酵条件下，通过离子交换色谱和反相高效液相色谱纯化后，从1升培养基中获得了纯度>95%的85.6毫克禽杀菌肽-2。重组禽杀菌肽-2表现出广谱抗菌活性，最低抑菌浓度为1至4μM。此外，重组禽杀菌肽-2表现出溶血活性，在128-256μM浓度范围内可促进50%的人红细胞溶血，还具有抗癌活性，在2-4μM浓度范围内可导致50%的A375人恶性黑色素瘤细胞死亡。本研究结果表明，重组禽杀菌肽-2可能是一种有前途的治疗应用候选物。
