Biotechnology Letters

, Volume 35, Issue 2, pp 233–238 | Cite as

Cloning and in silico characterization of two signal peptides from Pediococcus pentosaceus and their function for the secretion of heterologous protein in Lactococcus lactis

  • Ali Baradaran
  • Chin Chin Sieo
  • Hooi Ling Foo
  • Rosli Md. Illias
  • Khatijah Yusoff
  • Raha Abdul Rahim
Original Research Paper

Abstract

Fifty signal peptides of Pediococcus pentosaceus were characterized by in silico analysis and, based on the physicochemical analysis, (two potential signal peptides Spk1 and Spk3 were identified). The coding sequences of SP were amplified and fused to the gene coding for green fluorescent protein (GFP) and cloned into Lactococcus lactis pNZ8048 and pMG36e vectors, respectively. Western blot analysis indicated that the GFP proteins were secreted using both heterologous SPs. ELISA showed that the secretion efficiency of GFP using Spk1 (0.64 μg/ml) was similar to using Usp45 (0.62 μg/ml) and Spk3 (0.58 μg/ml).

Keywords

Green fluorescent protein Lactococcus lactis Secretion system Signal peptide 

Supplementary material

10529_2012_1059_MOESM1_ESM.doc (32 kb)
Supplementary material 1 (DOC 31 kb)
10529_2012_1059_MOESM2_ESM.doc (32 kb)
Supplementary material 2 (DOC 32 kb)
10529_2012_1059_MOESM3_ESM.doc (108 kb)
Supplementary material 3 (DOC 107 kb)

References

  1. Baradaran A, Foo HL, Sieo CC, Raha AR (2012) Isolation, identification and characterization of lactic acid bacteria from Polygonum minus. Rom Biotechnol Lett 17(2):7245–7252Google Scholar
  2. Bendtsen D (2004) Improved prediction of signal peptides: Signalp 3.0. J Mol Biol 340(4):783–795PubMedCrossRefGoogle Scholar
  3. Bolotin A, Wincker P, Mauger S, Jaillon O, Malarme K, Weissenbach J, Ehrlich SD, Sorokin A (2001) The complete genome sequence of the lactic acid bacterium Lactococcus lactis ssp. lactis IL1403. Gen Res 11(5):731–753CrossRefGoogle Scholar
  4. Brockmeier U, Caspers M, Freudl R, Jockwer A, Noll T, Eggert T (2006) Systematic screening of all signal peptides from Bacillus subtilis: a powerful strategy in optimizing heterologous protein secretion in Gram-positive bacteria. J Mol Biol 362(3):393–402PubMedCrossRefGoogle Scholar
  5. Choo K, Ranganathan S (2008) Flanking signal and mature peptide residues influence signal peptide cleavage. BMC Bioinformatics 9:S15. doi:10.1186/1471-2105-9-S12-S15 Google Scholar
  6. Desvaux M, Hébraud M, Talon R, Henderson I (2009) Secretion and subcellular localizations of bacterial proteins: a semantic awareness issue. Trends Microbiol 17(4):139–145PubMedCrossRefGoogle Scholar
  7. Gomi M, Sonoyama M, Mitaku S (2004) High performance system for signal peptide prediction: SOSUIsignal. Chem-Bio Inform J 4(4):142–147CrossRefGoogle Scholar
  8. Hiller K, Grote A, Scheer M, Munch R, Jahn D (2004) PrediSi: prediction of signal peptides and their cleavage positions. Nucleic Acid Res 32(Web Server Issue):W375–W379Google Scholar
  9. Le Loir Y, Nouaille S, Commissaire J, Bretigny L, Gruss A, Langella P (2001) Signal peptide and propeptide optimization for heterologous protein secretion in Lactococcus lactis. Appl Environ Microbiol 67(9):4119–4127PubMedCrossRefGoogle Scholar
  10. Le Loir Y, Azevedo V, Oliveira SC, Freitas DA, Miyoshi A, Bermudez-Humaran LG, Nouaille S, Ribeiro LA, Leclercq S, Gabriel JE, Guimaraes VD, Oliveira MN, Charlier C, Gautier M, Langella P (2005) Protein secretion in Lactococcus lactis: an efficient way to increase the overall heterologous protein production. Microb Cell Fact 4(1):2PubMedCrossRefGoogle Scholar
  11. Lindholm A, Smeds A, Palva A (2004) Receptor binding domain of Escherichia coli F18 fimbrial adhesin FedF can be both efficiently secreted and surface displayed in a functional form in Lactococcus lactis. Appl Environ Microbiol 70:2061–2071PubMedCrossRefGoogle Scholar
  12. Morello E, Bermdez-Humarn L, Llull D, Solé V, Miraglio N, Langella P, Poquet I (2008) Lactococcus lactis, an efficient cell factory for recombinant protein production and secretion. J Mol Microbiol Biotechnol 14(1–3):48–58PubMedCrossRefGoogle Scholar
  13. Sriraman K, Jayaraman G (2006) Enhancement of recombinant streptokinase production in Lactococcus lactis by suppression of acid tolerance response. Appl Microbiol Biotechnol 72(6):1202–1209PubMedCrossRefGoogle Scholar
  14. Tremillon N, Issaly N, Mozo J, Duvignau T, Ginisty H, Devic E, Poquet E (2010) Production and purification of staphylococcal nuclease in Lactococcus lactis using a new expression-secretion system and a pH-regulated mini-reactor. Microb Cell Fact 9(1):37. doi:10.1186/1475-2859-9-37 PubMedCrossRefGoogle Scholar
  15. Villatoro-Hernandez J, Loera-Arias M, Gamez-Escobedo A, Franco-Molina M et al (2008) Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis. Microb Cell Fact 7:22–32PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Ali Baradaran
    • 1
  • Chin Chin Sieo
    • 2
    • 4
  • Hooi Ling Foo
    • 3
    • 4
  • Rosli Md. Illias
    • 5
  • Khatijah Yusoff
    • 2
    • 4
  • Raha Abdul Rahim
    • 1
    • 4
  1. 1.Department of Cell and Molecular BiologyFaculty of Biotechnology and Biomolecular Sciences, Universiti Putra MalaysiaSerdangMalaysia
  2. 2.Department of MicrobiologyFaculty of Biotechnology and Biomolecular Sciences, Universiti Putra MalaysiaSerdangMalaysia
  3. 3.Department of Bioprocess TechnologyFaculty of Biotechnology & Biomolecular Sciences, Universiti Putra MalaysiaUPM, SerdangMalaysia
  4. 4.Institute of BioscienceUPM, SerdangMalaysia
  5. 5.Department of Bioprocess EngineeringFaculty of Chemical Engineering, Universiti Teknologi MalaysiaSkudaiMalaysia

Personalised recommendations