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Expression and Purification of an Antimicrobial Peptide by Fusion with Elastin-like Polypeptides in Escherichia coli

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Abstract

Different carrier molecules have been fused to antimicrobial polypeptides (AMPs) to facilitate recombinant protein expression and purification. Some of them have improved the stability of AMPs and reduced the toxicity to host cells, but most current strategies still have some problems to be solved such as poor yield, low purity, high expense, time-consumption, and difficulty in scaling-up. Here, we introduced the elastin-like polypeptides (ELPs) as a fusion partner to express an antimicrobial polypeptide halocidin18 (Hal18). By the reversible soluble–insoluble phase transition, 69 mg of the fusion protein were purified from 1 l of culture medium with the purity of nearly 95%. After cleavage with hydroxylamine, the ELP’s tag was easily separated from Hal18 in the next round of inverse transition cycle and Hal18 (1.7 mg, ∼1.9 kDa) was mainly found in the supernatant with a recovery of about 47% and purity of 60%. Antimicrobial activity showed that Hal18 had strong antimicrobial activity against Escherichia coli and Micrococcus luteus but weak activity against Pichia pastoris.

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Acknowledgments

This work was supported by the National Natural Science Fund of China (number 30760009), the grants from the China National Human Liver Proteomics Project (2004BA711A19) and the China National High-Tech 863 Program (2006AA02A310), the Fund of Nanyang Normal University (number nynu200748). The authors thank Prof. Xing Guo Wang (Department of Nuclear Medicine, Albert Einstein College of Medicine, United States of America) and Prof. W.J. Cram for their valuable suggestions and corrections to the manuscript.

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Authors and Affiliations

  1. College of Life Science, Hubei University, Wuhan, 430062, China

    Fan Hu, Xin Li, Xiang Dong Ma & Li Xin Ma

  2. College of Life Science and Technology, Nanyang Normal University, Nanyang, 473061, China

    Tao Ke & Feng Li Hui

  3. Institute of Ion Beam Biotechnology, College of Physics Science and Technology, Xinjiang University, Urumqi, Xinjiang, 830008, China

    Pei Hong Mao & Xiang Jin

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  1. Fan Hu
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  2. Tao Ke
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  3. Xin Li
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  5. Xiang Jin
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  6. Feng Li Hui
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Correspondence to Xiang Dong Ma.

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Fan Hu and Tao Ke contributed equally to this work.

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Hu, F., Ke, T., Li, X. et al. Expression and Purification of an Antimicrobial Peptide by Fusion with Elastin-like Polypeptides in Escherichia coli . Appl Biochem Biotechnol 160, 2377–2387 (2010). https://doi.org/10.1007/s12010-009-8850-2

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  • DOI: https://doi.org/10.1007/s12010-009-8850-2

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