Abstract
The abuse of antibiotics has led to the emergence of many resistant bacteria, which threaten public health on a global scale. Conventional pipelines for antibiotic discovery are gradually exhausted. Here a combinatorial yeast overlay method is presented for the isolation of antibacterial oligopeptides. Three antibacterial dipeptide and tripeptides were discovered from 20 attempts, two of which possess glycosylation sites. Although the antibacterial effects were modest against several tested bacteria including a multi-resistant Staphylococcus aureus strain Y5, they are amenable to chemical modifications to enhance potency. This cost-effective method can circumvent the expensive chemical peptide synthesis process. The eukaryotic host Saccharomyces cerevisiae adopted for peptide expression could serve as a surrogate biosafety assay system for antibacterial peptides. The small size of the oligopeptides render them attractive candidates for drug development. Given the vast combinatorial diversity of peptides, the reservoir may serve as a rich source of drug leads.
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Acknowledgments
This work was supported by Guangdong Natural Science Foundation of China (S2011010004264, 05003328), Guangdong Science and Technology Program (No. 2008B020100001), National Natural Science Foundation of China (No. 30370799), Open Fund of MOE Key Laboratory of Aquatic Product Safety, Open Fund of Laboratory at Sun Yat-sen University (KF201231), Wen’s Group (wens-kjhz-2007-03-09-01), and The Genetics Teaching Group Project of Guangdong Province. Valuable suggestions from Dr. Wenqing Zhang and Dr. Yuncan Ai are appreciated. The authors acknowledge the contributions from Li Yu, Li Xu, Chujing Zhang, Jiashou Fang, Yuan Wang and Li Chen, and the authors are grateful to Mr. Hans E. Seidel and Ms. Yan Shi for proof reading and anonymous reviewers for valuable suggestions. The standard S. aureus strain is ATCC6538. P. aeruginosa strain 1.2464 was purchased from China General Microbiological Culture Collection Center (CGMCC, Beijing, China). E. coli strain MG1655 was a gift of the Coli Genetic Stock Center at Yale University. E. coli strain DH5α was obtained from Promega (Madison, WI, USA). The Y5 S. aureus strain was collected by Guangzhou Center for Disease Control and Prevention, which is resistant to ampicillin, penicillin, erythromycin and norfloxacin classified according to NCCLS2005 recommendations. It is also moderately sensitive to clindamycin and nitrofurantoin. E. tarda and A. hydrophila strains were the generous gift of Pearl River Fisheries Research Institute. S. strain S150-2B was a kind gift of Dr. Derek Jamieson of Heriot-Watt University, United Kingdom.
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Ran He, Liang Cui, Yan Cao, Zhixue Wang, Kai Wang and Min Fu contributed equally to this work.
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He, R., Cui, L., Cao, Y. et al. A Combinatorial Yeast Overlay Method for the Isolation of Antibacterial Oligopeptides. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 84, 1069–1075 (2014). https://doi.org/10.1007/s40011-013-0257-8
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DOI: https://doi.org/10.1007/s40011-013-0257-8