Antimicrobial Peptides from Musca Domestica Expressed in Escherichia coli

Abstract:

Cecropins are cationic molecules with a wide range of antimicrobial activities. The native peptide cecropins from Musca domestica (Md-Cec) have antimicrobial activity against both Gram-positive and Gram-negative bacteria. In the present study, cDNA fragments encoding both the Md-Cec-L (62aa) and Md-Cec-S (40 aa) peptides of Md-Cec were respectively expressed using the pMAL-c4x expression vector. High level expression of Md-Cec-L was achieved in Escherichia coli, while expression of Md-Cec-S failed to reach a decent level due to its high level of toxicity to the host cells. Md-Cec-L was expressed as a soluble form using a maltose binding protein (MBP) system, whose product is a MBP-tagged fusion protein, and separated with the carrier amyrose resin. Heterologous expression in E. coli and antimicrobial activity assays showed that both the recombinant fusion protein Md-Cec-L and Md-Cec-S have exhibited antimicrobial activity in vivo; and Md-Cec-L also exhibited antimicrobial activity in vitro. Md-Cec has the potential to be developed as a novel type of antimicrobial drug or food preservative.

• Periodical: Advanced Materials Research (Volumes 535-537)
• Pages: 2404-2408
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.535-537.2404
• Online since: June 2012
• Authors: Bin Zeng
• Keywords: Antimicrobial Activity, Fusion Protein Expression, Musca domestica Cecropins

References

[1] Jiang SJ. Chinese Pharmaceutical Insects. Beijing, China: Chinese Forestry Publishing House (1999).

[2] Hou L, Shi Y, Zhai P and Le G. Antibacterial activity and in vitro antitumor activity of the extract of the larvae of the housefly (Musca domestica). J Ethnopharmacol 111 ( 2007), p.227–231.

DOI: 10.1016/j.jep.2006.11.015

[3] Boman HG. Peptide antibiotics and their role in innate immunity. Ann Rev Immunol 13 (1995), p.61–92.

DOI: 10.1146/annurev.iy.13.040195.000425

[4] Bulet P, Hetru C, Dimarcq JL and Hoffmann D. Antimicrobial peptides in insects; structure and function. Dev Comp Immunol 23 (1999), p.329–344.

DOI: 10.1016/s0145-305x(99)00015-4

[5] Liang Y, Wang JX, Zhao XF, Du XJ and Xue JF. Molecular cloning and characterization of cecropin from the housefly (Musca domestica), and its expression in Escherichia coli. Dev Comp Immunol 30 (2006), p.249–257.

DOI: 10.1016/j.dci.2005.04.005

[6] Kisich KO,Howell M D,Boguniewicz M, Heizer H R, Watson N U, Leung DY: The constitutive capacity of human keratinocytes to ill Staphylococcus aureus is dependent on beta-defensin3. J. Invest Dermatol 127 (2007), pp.2368-2380

DOI: 10.1038/sj.jid.5700861

[7] Liu PT,Modlin RL Human macrophage host defense against Mycobacterium tuberculosis. Curr Opin Immunol 20 (2008), pp.371-376

DOI: 10.1016/j.coi.2008.05.014

[8] Lu XM, Jin XB, Zhu JY, Mei HF, Ma Y, Chu FJ, Wang Y, Li XB. Expression of the antimicrobial peptide cecropin fused with human lysozyme in Escherichia coli. Appl Microbiol Biotechnol. 87 (2010), pp.2169-76

DOI: 10.1007/s00253-010-2606-3