Journal
EXPERIMENTAL AND THERAPEUTIC MEDICINE
Volume 12, Issue 4, Pages 2324-2330Publisher
SPANDIDOS PUBL LTD
DOI: 10.3892/etm.2016.3578
Keywords
antimicrobial peptides; fowlicidin-2; Pichia pastoris; recombinant expression; antibacterial activity; anticancer activity
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Funding
- Program for Young Aged Academic Staff in the Heilongjiang Province Ordinary College [1252G010)U]
- Research Fund for Innovation Talents of Science and Technology in Harbin City [2012RFQXN022]
- 'Academic Backbone' Project of Northeast Agricultural University [15XG15]
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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 Pichia pastoris X-33 and the expression vector pPICZ-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 mu M. Furthermore, recombinant fowlicidin-2 exhibited hemolytic activity, promoting 50% human erythrocyte hemolysis in the concentration range of 128-256 mu M, and anticancer activity, resulting in the death of 50% of A375 human malignant melanoma cells in the concentration range of 2-4 mu M. The results of the present study suggest that recombinant fowlicidin-2 may be a promising candidate for therapeutic applications.
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