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Expression and purification of antimicrobial peptide buforin IIb in Escherichia coli

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Abstract

A novel production method in Escherichia coli for an antimicrobial peptide of 21 amino acids, buforin IIb, which is a synthetic analog of buforin II, has been developed. The buforin IIb gene was cloned into the vector pET32a to construct an expression vector pET32a–buforin IIb. The fusion protein Trx-buforin IIb, purified by nickel nitrilo-triacetic acid (Ni-NTA) resin chromatography, was cleaved by hydroxylamine hydrochloride to release recombinant buforin IIb. Purification of recombinant buforin IIb was achieved by HPLC: about 3.1 mg/l active recombinant buforin IIb with purity >99% was obtained. The recombinant buforin IIb showed antimicrobial activities that were similar to the synthetic one.

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Acknowledgments

This work was supported by the special funding from NJNU for talent faculty and Jiangsu superior discipline.

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Authors and Affiliations

  1. Jiangsu Province Key Laboratory for Molecular and Medicine Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, No. 1 Wenyuan Road, Nanjing, 210046, Jiangsu, People’s Republic of China

    Qi Wang, Fenfen Zhu, Yinqiang Xin, Jia Liu & Zhimin Yin

  2. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, 210093, People’s Republic of China

    Lan Luo

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  1. Qi Wang
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  2. Fenfen Zhu
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  3. Yinqiang Xin
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  4. Jia Liu
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  5. Lan Luo
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  6. Zhimin Yin
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Correspondence to Lan Luo or Zhimin Yin.

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Wang, Q., Zhu, F., Xin, Y. et al. Expression and purification of antimicrobial peptide buforin IIb in Escherichia coli . Biotechnol Lett 33, 2121–2126 (2011). https://doi.org/10.1007/s10529-011-0687-4

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  • DOI: https://doi.org/10.1007/s10529-011-0687-4

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