Applied Microbiology and Biotechnology

, Volume 98, Issue 8, pp 3651–3658 | Cite as

Expressing antimicrobial peptide cathelicidin-BF in Bacillus subtilis using SUMO technology

  • Chao Luan
  • Hai Wen Zhang
  • De Guang Song
  • Yong Gang Xie
  • Jie Feng
  • Yi Zhen WangEmail author
Biotechnologically relevant enzymes and proteins


Small ubiquitin-related modifier (SUMO) technology has been widely used in Escherichia coli expression systems to produce antimicrobial peptides. However, E. coli is a pathogenic bacterium that produces endotoxins and can secrete proteins into the periplasm, forming inclusion bodies. In our work, cathelicidin-BF (CBF), an antimicrobial peptide purified from Bungarus fasciatus venom, was produced in a Bacillus subtilis expression system using SUMO technology. The chimeric genes his-SUMO-CBF and his-SUMO protease 1 were ligated into vector pHT43 and expressed in B. subtilis WB800N. Approximately 22 mg of recombinant fusion protein SUMO-CBF and 1 mg of SUMO protease 1 were purified per liter of culture supernatant. Purified SUMO protease 1 was highly active and cleaved his-SUMO-CBF with an enzyme-to-substrate ratio of 1:40. Following cleavage, recombinant CBF was further purified by affinity and cation exchange chromatography. Peptide yields of ~3 mg/l endotoxin-free CBF were achieved, and the peptide demonstrated antimicrobial activity. This is the first report of the production of an endotoxin-free antimicrobial peptide, CBF, by recombinant DNA technology, as well as the first time purified SUMO protease 1 with high activity has been produced from B. subtilis. This work has expanded the application of SUMO fusion technology and may represent a safe and efficient way to generate peptides and proteins in B. subtilis.


Small ubiquitin-related modifier SUMO protease 1 Bacillus subtilis Cathelicidin-BF Antimicrobial peptide 



This work was financially supported by the Modern Agro-Industry Technology Research System (no. CARS-36) and 12th Five-year Plan-National Science and Technology Programs in Agricultural Areas (2011BAD26B002-5).


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Chao Luan
    • 1
  • Hai Wen Zhang
    • 1
  • De Guang Song
    • 1
  • Yong Gang Xie
    • 1
  • Jie Feng
    • 1
  • Yi Zhen Wang
    • 1
    Email author
  1. 1.Key Laboratory of Animal Nutrition and Feed Science, Ministry of Agriculture (East China), Zhejiang Provincial Laboratory of Feed and Animal Nutrition, Institute of Feed ScienceZhejiang UniversityHangzhouPeople’s Republic of China

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