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Article

Expression of cecropin B in Pichia pastoris and its bioactivity in vitro

  • Authors:
    • Xiuqing Wang
    • Mingxing Zhu
    • Guimao Yang
    • Chunxia Su
    • Aijun Zhang
    • Ruibing Cao
    • Puyan Chen
  • View Affiliations / Copyright

    Affiliations: School of Clinical Laboratory of Medicine, NingXia Medical University, Yinchuan 750004, P.R. China, Laboratory Animal Centre, NingXia Medical University, Yinchuan 750004, P.R. China, School of Basic Medical Science, NingXia Medical University, Yinchuan 750004, P.R. China, Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, P.R. China
  • Pages: 655-660
    |
    Published online on: May 9, 2011
       https://doi.org/10.3892/etm.2011.262
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Abstract

Natural cecropin B (CB), consisting of 35 amino acids, is a member of the cecropin family with the highest antibacterial activity. Here, a novel approach for the generation of recombinant CB in the methylotrophic yeast Pichia pastoris was explored. For this purpose, the CB gene was amplified by recursive PCR (rPCR) and cloned into the pPICZαA vector. The SacI-linearized plasmid pPICZαA-CB was transformed into P. pastoris SMD1168 by electroporation. The expression of recombinant CB was induced with 1.0% methanol at pH 5.0 for 60 h at 28˚C. Recombinant CB was purified using cationic exchange chromatography; 5.0 mg of pure active CB was obtained from 100 ml of culture broth supernatant. Antimicrobial assays demonstrated that CB has a broad spectrum of antimicrobial activity against both Gram-positive and Gram-negative bacteria. Our results suggest that the P. pastoris expression system can be used to produce large quantities of fully functional CB for both research and industrial purposes.
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1. 

Devine DA and Hancock RE: Cationic peptides: distribution and mechanisms of resistance. Curr Pharm Des. 8:703–714. 2002. View Article : Google Scholar : PubMed/NCBI

2. 

Ganz T: Defensins: antimicrobial peptides of innate immunity. Nat Rev Immunol. 3:710–720. 2003. View Article : Google Scholar : PubMed/NCBI

3. 

Segrest JP, De Loof H, Dohlman JG, Brouillette CG and Anantharamaiah GM: Amphipathic helix motif: classes and properties. Proteins. 8:103–117. 1990. View Article : Google Scholar : PubMed/NCBI

4. 

Zasloff M: Antimicrobial peptides of multicellular organisms. Nature. 415:389–395. 2002. View Article : Google Scholar : PubMed/NCBI

5. 

Boman HG: Peptide antibiotics and their role in innate immunity. Annu Rev Immunol. 13:61–92. 1995. View Article : Google Scholar : PubMed/NCBI

6. 

Noga EJ and Silphaduang U: Piscidins: a novel family of peptide antibiotics from fish. Drug News Perspect. 16:87–92. 2003. View Article : Google Scholar : PubMed/NCBI

7. 

Steiner H, Hultmark D, Engstrom A, Bennich H and Boman HG: Sequence and specificity of two antibacterial proteins involved in insect immunity. Nature. 292:246–248. 1981. View Article : Google Scholar

8. 

Boman HG and Hultmark D: Cell-free immunity in insects. Annu Rev Microbiol. 41:103–126. 1987. View Article : Google Scholar : PubMed/NCBI

9. 

Lee JY, Boman A, Sun CX, Andersson M, Jornvall H, Mutt V and Boman HG: Antibacterial peptides from pig intestine: isolation of a mammalian cecropin. Proc Natl Acad Sci USA. 86:9159–9162. 1989. View Article : Google Scholar : PubMed/NCBI

10. 

Hultmark D, Engstrom A, Bennich H, Kapur R and Boman HG: Insect immunity: isolation and structure of cecropin D and four minor antibacterial components from Cecropia pupae. Eur J Biochem. 127:207–217. 1982. View Article : Google Scholar : PubMed/NCBI

11. 

Boman HG, Wade D, Boman IA, Wahlin B and Merrifield RB: Antibacterial and antimalarial properties of peptides that are cecropin-melittin hybrids. FEBS Lett. 259:103–106. 1989. View Article : Google Scholar : PubMed/NCBI

12. 

Moore AJ, Devine DA and Bibby MC: Preliminary experimental anticancer activity of cecropins. Pept Res. 7:265–269. 1994.PubMed/NCBI

13. 

Hancock RE and Lehrer R: Cationic peptides: a new source of antibiotics. Trends Biotechnol. 16:82–88. 1998.PubMed/NCBI

14. 

Hancock RE and Patrzykat A: Clinical development of cationic antimicrobial peptides: from natural to novel antibiotics. Curr Drug Targets Infect Disord. 2:79–83. 2002. View Article : Google Scholar : PubMed/NCBI

15. 

Diamond G: Natures antibiotics: the potential of antimicrobial peptides as new drugs. Biologist. 48:209–212. 2001.PubMed/NCBI

16. 

Andres E and Dimarcq JL: Cationic antimicrobial peptides: from innate immunity study to drug development. Med Mal Infect. 37:194–199. 2007.PubMed/NCBI

17. 

Cereghino JL and Cregg JM: Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev. 24:45–66. 2000. View Article : Google Scholar

18. 

Sreekrishna K, Brankamp RG, Kropp KE, Blankenship DT, Tsay JT, Smith PL, Wierschke JD, Subramaniam A and Birkenberger LA: Strategies for optimal synthesis and secretion of heterologous proteins in the methylotrophic yeast Pichia pastoris. Gene. 190:55–62. 1997. View Article : Google Scholar : PubMed/NCBI

19. 

Cregg JM, Cereghino JL, Shi J and Higgins DR: Recombinant protein expression in Pichia pastoris. Mol Biotechnol. 16:23–52. 2000. View Article : Google Scholar : PubMed/NCBI

20. 

Boze H, Celine L, Patrick C, Fabien R, Christine V, Yves C and Guy M: High-level secretory production of recombinant porcine follicle-stimulating hormone by Pichia pastoris. Process Biochem. 36:907–913. 2001. View Article : Google Scholar

21. 

Lee CY, Lee SJ, Jung KH, Katoh S and Lee K: High dissolved oxygen tension enhances heterologous protein expression by recombinant Pichia pastoris. Process Biochem. 38:1147–1154. 2003. View Article : Google Scholar

22. 

Almeida MS, Cabral KS, de Medeiros LN, Valente AP, Almeida FC and Kurtenbach E: cDNA cloning and heterologous expression of functional cysteine-rich antifungal protein Psd1 in the yeast Pichia pastoris. Arch Biochem Biophys. 395:199–207. 2001. View Article : Google Scholar : PubMed/NCBI

23. 

Ausubel FM, Brent R and Kingston RE: Protocols in molecular biology. Trends Cell Biol. 6:366–367. 1996.

24. 

Lehrer RI, Rosenman M, Harwig SS, Jackson R and Eisenhauer P: Ultrasensitive assays for endogenous peptides. J Immunol Methods. 137:167–173. 1991. View Article : Google Scholar

25. 

Cipakova I, Hostinova E, Gasperik J and Velebny V: High-level expression and purification of a recombinant hBD-1 fused to LMM protein in Escherichia coli. Protein Expr Purif. 37:207–212. 2004. View Article : Google Scholar : PubMed/NCBI

26. 

Shi X, Karkut T, Chamankhah M, Alting-Mees M, Hemmingsen SM and Hegedus D: Optimal conditions for the expression of a single-chain antibody (scFv) gene in Pichia pastoris. Protein Express Purif. 28:321–330. 2003. View Article : Google Scholar : PubMed/NCBI

27. 

Jahic M, Gustavsson M, Jansen AK, Martinelle M and Enfors SO: Analysis and control of proteolysis of a fusion protein in Pichia pastoris fed-batch processes. J Biotechnol. 102:45–53. 2003. View Article : Google Scholar : PubMed/NCBI

28. 

Li SZ, Xiong F and Lin Q: Low-temperature increases the yield of biologically active herring antifreeze protein in Pichia pastoris. Protein Express Purif. 21:438–445. 2001. View Article : Google Scholar : PubMed/NCBI

29. 

Bencurova M, Rendic D, Fabini G, Kopecky EM, Altmann F and Wilson IB: Expression of eukaryotic glycosyltransferases in the yeast Pichia pastoris. Biochimie. 85:413–422. 2003. View Article : Google Scholar : PubMed/NCBI

30. 

Skosyrev VS, Kulesskiy EA, Yakhnin AV, Temirov YV and Vinokurov LM: Expression of the recombinant antibacterial peptide sarcotoxin IA in Escherichia coli cells. Protein Expr Purif. 28:350–356. 2003. View Article : Google Scholar : PubMed/NCBI

31. 

Aly R, Granot D, Mahler-Slasky Y, Halpern N, Nir D and Galun E: Saccharomyces cerevisiae cells harboring the gene encoding sarcotoxin IA secrete a peptide that is toxic to plant pathogenic bacteria. Protein Expr Purif. 16:120–124. 1999. View Article : Google Scholar

32. 

Yamada K, Nakajima Y and Nator S: Production of recombinant sarcotoxin IA in Bombyx mori cells. Biochem J. 272:633–636. 1990.PubMed/NCBI

33. 

Rao XC, Li S, Hu JC, Jin XL, Hu XM, Huang JJ, Chen ZJ, Zhu JM and Hu FQ: A novel carrier molecule for high-level expression of peptide antibiotics in Escherichia coli. Protein Expr Purif. 36:11–18. 2004. View Article : Google Scholar : PubMed/NCBI

34. 

Cabral KM, Almeida MS, Valente AP, Almeida FC and Kurtenbach E: Production of the active antifungal Pisum sativum defensin 1 (Psd1) in Pichia pastoris: overcoming the inefficiency of the STE13 protease. Protein Expr Purif. 31:115–122. 2003.PubMed/NCBI

35. 

Sarmasik A and Chen TT: Bactericidal activity of cecropin B and cecropin P1 expressed in fish cells (CHSE-214): application in controlling fish bacterial pathogens. Aquaculture. 220:183–194. 2003. View Article : Google Scholar

36. 

Sessitsch A, Kan FY and Pfeifer U: Diversity and community structure of culturable Bacillus spp. populations in the rhizospheres of transgenic potatoes expressing the lytic peptide cecropin B. Appl Soil Ecol. 22:149–158. 2003.

37. 

Sharma A, Sharma R, Imamura M, Yamakawa M and Machii H: Transgenic expression of cecropin B, an antibacterial peptide from Bombyx mori, confers enhanced resistance to bacterial leaf blight in rice. FEBS Lett. 484:7–11. 2000. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Wang X, Zhu M, Yang G, Su C, Zhang A, Cao R and Chen P: Expression of cecropin B in Pichia pastoris and its bioactivity in vitro. Exp Ther Med 2: 655-660, 2011.
APA
Wang, X., Zhu, M., Yang, G., Su, C., Zhang, A., Cao, R., & Chen, P. (2011). Expression of cecropin B in Pichia pastoris and its bioactivity in vitro. Experimental and Therapeutic Medicine, 2, 655-660. https://doi.org/10.3892/etm.2011.262
MLA
Wang, X., Zhu, M., Yang, G., Su, C., Zhang, A., Cao, R., Chen, P."Expression of cecropin B in Pichia pastoris and its bioactivity in vitro". Experimental and Therapeutic Medicine 2.4 (2011): 655-660.
Chicago
Wang, X., Zhu, M., Yang, G., Su, C., Zhang, A., Cao, R., Chen, P."Expression of cecropin B in Pichia pastoris and its bioactivity in vitro". Experimental and Therapeutic Medicine 2, no. 4 (2011): 655-660. https://doi.org/10.3892/etm.2011.262
Copy and paste a formatted citation
x
Spandidos Publications style
Wang X, Zhu M, Yang G, Su C, Zhang A, Cao R and Chen P: Expression of cecropin B in Pichia pastoris and its bioactivity in vitro. Exp Ther Med 2: 655-660, 2011.
APA
Wang, X., Zhu, M., Yang, G., Su, C., Zhang, A., Cao, R., & Chen, P. (2011). Expression of cecropin B in Pichia pastoris and its bioactivity in vitro. Experimental and Therapeutic Medicine, 2, 655-660. https://doi.org/10.3892/etm.2011.262
MLA
Wang, X., Zhu, M., Yang, G., Su, C., Zhang, A., Cao, R., Chen, P."Expression of cecropin B in Pichia pastoris and its bioactivity in vitro". Experimental and Therapeutic Medicine 2.4 (2011): 655-660.
Chicago
Wang, X., Zhu, M., Yang, G., Su, C., Zhang, A., Cao, R., Chen, P."Expression of cecropin B in Pichia pastoris and its bioactivity in vitro". Experimental and Therapeutic Medicine 2, no. 4 (2011): 655-660. https://doi.org/10.3892/etm.2011.262
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