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Expression and purification of lacticin Q by small ubiquitin-related modifier fusion in Escherichia coli

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Abstract

Lacticin Q is a broad-spectrum class II bacteriocin with potential as an alternative to conventional antibiotics. The objective of this study was to produce recombinant lacticin Q using a small ubiquitin-related modifier (SUMO) fusion protein expression system. The 168-bp lacticin Q gene was cloned into the expression vector pET SUMO and transformed into Escherichia coli BL21(DE3). The soluble fusion protein was recovered with a Ni-NTA Sepharose column (95% purity); 130 mg protein was obtained per liter of fermentation culture. The SUMO tag was then proteolytically cleaved from the protein, which was re-applied to the column. Finally, about 32 mg lacticin Q (≥96% purity) was obtained. The recombinant protein exhibited antimicrobial properties similar to that of the native protein, demonstrating that lacticin Q had been successfully expressed by the SUMO fusion system.

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References

  • Beaulieu, L., Groleau, D., Miguez, C.B., Jetté, J.F., Aomari, H., and Subirade, M. 2005. Production of pediocin PA-1 in the methylotrophic yeast Pichia pastoris reveals unexpected inhibition of its biological activity due to the presence of collagen-like material. Protein Expr. Purif. 43, 111–125.

    Article  PubMed  CAS  Google Scholar 

  • Bommarius, B., Jenssen, H., Elliott, M., Kindrachuk, J., Pasupuleti, M., Gieren, H., Jaeger, K.E., Hancock, R.E.W., and Kalman, D. 2010. Cost-effective expression and purification of antimicrobial and host defense peptides in Escherichia coli. Peptides 31, 1957–1965.

    Article  PubMed  CAS  Google Scholar 

  • Butt, T.R., Edavettal, S.C., Hall, J.P., and Mattern, M.R. 2005. SUMO fusion technology for difficult-to-express proteins. Protein Expr. Purif. 43, 1–9.

    Article  PubMed  CAS  Google Scholar 

  • Čipáková, I., Hostinová, E., Gašperík, J., and Velebný, V. 2004. High-level expression and purification of a recombinant hBD-1 fused to LMM protein in Escherichia coli. Protein Expr. Purif. 37, 207–212.

    Article  PubMed  Google Scholar 

  • Fujita, K., Ichimasa, S., Zendo, T., Koga, S., Yoneyama, F., Nakayama, J., and Sonomoto, K. 2007. Structural analysis and characterization of lacticin Q, a novel bacteriocin belonging to a new family of unmodified bacteriocins of Gram-positive bacteria. Appl. Environ. Microbiol. 73, 2871–2877.

    Article  PubMed  CAS  Google Scholar 

  • Gillor, O. and Ghazaryan, L. 2007. Recent advances in bacteriocin application as antimicrobials. Recent Pat. Anti-Infect. Drug Discovery 2, 115–122.

    Article  CAS  Google Scholar 

  • Jasniewski, J., Cailliez-Grimal, C., Gelhaye, E., and Revol-Junelles, A.M. 2008. Optimization of the production and purification processes of carnobacteriocins Cbn BM1 and Cbn B2 from Carnobacterium maltaromaticum CP5 by heterologous expression in Escherichia coli. J. Microbiol. Methods 73, 41–48.

    Article  PubMed  CAS  Google Scholar 

  • Lee, D.G., Park, Y., Kim, H.N., Kim, H.K., Kim, P.I., Choi, B.H., and Hahm, K.-S. 2002. Antifungal mechanism of an antimicrobial peptide, HP (2–20), derived from N-terminus of Helicobacter pylori ribosomal protein L1 against Candida albicans. Biochem. Biophys. Res. Commun. 291, 1006–1013.

    Article  PubMed  CAS  Google Scholar 

  • Li, Y. 2009. Carrier proteins for fusion expression of antimicrobial peptides in Escherichia coli. Biotechnol. Appl. Biochem. 54, 1–9.

    Article  PubMed  CAS  Google Scholar 

  • Li, J.F., Zhang, J., Song, R., Zhang, J.X., Shen, Y., and Zhang, S.Q. 2009. Production of a cytotoxic cationic antibacterial peptide in Escherichia coli using SUMO fusion partner. Appl. Microbiol. Biotechnol. 84, 383–388.

    Article  PubMed  CAS  Google Scholar 

  • Li, J.F., Zhang, J., Zhang, Z., Ma, H.W., Zhang, J.X., and Zhang, S.Q. 2010. Production of bioactive human beta-defensin-4 in Escherichia coli using SUMO fusion partner. Protein J. 29, 314–319.

    Article  PubMed  CAS  Google Scholar 

  • Morin, K.M., Arcidiacono, S., Beckwitt, R., and Mello, C.M. 2006. Recombinant expression of indolicidin concatamers in Escherichia coli. Appl. Microbiol. Biotechnol. 70, 698–704.

    Article  PubMed  CAS  Google Scholar 

  • Muller, S., Hoege, C., Pyrowolakis, G., and Jentsch, S. 2001. Sumo, ubiquitin’s mysterious cousin. Nat. Rev. Mol. Cell Biol. 2, 202–210.

    Article  PubMed  CAS  Google Scholar 

  • Rao, X.C., Li, S., Hu, J.C., Jin, X.L., Hu, X.M., Huang, J.J., Chen, Z.J., Zhu, J.M., and Hu, F.Q. 2004. A novel carrier molecule for high-level expression of peptide antibiotics in Escherichia coli. Protein Expr. Purif. 36, 11–18.

    Article  PubMed  CAS  Google Scholar 

  • Riley, M.A. and Wertz, J.E. 2002. Bacteriocin diversity: Ecological and evolutionary perspectives. Biochimie 84, 357–364.

    Article  PubMed  CAS  Google Scholar 

  • Schagger, H. 2006. Tricine-SDS-PAGE. Nat. Protoc. 1, 16–22.

    Article  PubMed  Google Scholar 

  • Shen, Y., Ai, H.X., Song, R., Liang, Z.N., Li, J.F., and Zhang, S.Q. 2010. Expression and purification of moricin CM4 and human β-defensins 4 in Escherichia coli using a new technology. Microbiol. Res. 165, 713–718.

    Article  PubMed  CAS  Google Scholar 

  • Sun, Z., Xia, Z., Bi, F., and Liu, J.N. 2008. Expression and purification of human urodilatin by small ubiquitin-related modifier fusion in Escherichia coli. Appl. Microbiol. Biotechnol. 78, 495–502.

    Article  PubMed  CAS  Google Scholar 

  • Torkar, K.G. and Bogovi, M.B. 2003. Partial characterisation of bacteriocins produced by Bacillus cereus isolates from milk and milk products. Food Technol. Biotechnol. 41, 121–129.

    CAS  Google Scholar 

  • Wei, Q., Kim, Y.S., Seo, J.H., Jang, W.S., Lee, I.H., and Cha, H.J. 2005. Facilitation of expression and purification of an antimicrobial peptide by fusion with baculoviral polyhedrin in Escherichia coli. Appl. Environ. Microbiol. 71, 5038–5043.

    Article  PubMed  CAS  Google Scholar 

  • Xie, J., Zhang, R., Shang, C., and Guo, Y. 2009. Isolation and characterization of a bacteriocin produced by an isolated Bacillus subtilis LFB112 that exhibits antimicrobial activity against domestic animal pathogens. Afr. J. Biotechnol. 8, 5611–5619.

    Google Scholar 

  • Yu, F., Wang, J., Zhang, P., Hong, Y., and Liu, W. 2010. Fusion expression of cecropin B-like antibacterial peptide in Escherichia coli and preparation of its antiserum. Biotechnol. Lett. 32, 669–673.

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Rijun Zhang.

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These authors contributed equally to this work.

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Ma, Q., Yu, Z., Han, B. et al. Expression and purification of lacticin Q by small ubiquitin-related modifier fusion in Escherichia coli . J Microbiol. 50, 326–331 (2012). https://doi.org/10.1007/s12275-012-1425-x

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  • DOI: https://doi.org/10.1007/s12275-012-1425-x

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