Skip to main content
SpringerLink
Log in

Expression of Stichopus japonicus Lysozyme Gene in Bacillus subtilis WB600

  • Conference paper
  • First Online:
Advances in Applied Biotechnology

Part of the book series: Lecture Notes in Electrical Engineering ((LNEE,volume 332))

  • 1804 Accesses

Abstract

In this study, a genetic engineering bacteria Bacillus subtilis pHT43-SjLys/WB600 was successfully constructed for the expression of the lysozyme gene from sea cucumber (Stichopus japonicus) by the method of recombinant DNA technique. The growth trend of engineering bacteria was consistent with the wild-type strain WB600, and the results demonstrated that insertion of foreign gene did not affect its physiological and biochemical metabolism. In the absence of selection pressure, the analysis of the stability revealed that there was no gene rearrangement and lost of the recombinant plasmid in the bacteria which showed that it has high genetic stability. The SDS-PAGE results demonstrated that pHT43-SjLys/WB600 successfully expressed soluble SjLys protein after incubated for 48 h induced by ITPG. The heterologous expression protein of pHT43-SjLys/WB600 displayed remarkable inhibitive effect on the growth of the Vibrio parahaemolyticus. To our knowledge, this is the first report about the SjLys gene authentic heterologous expression in B. subtilis. It should provide a robust secretion expression system for genetic engineering of B. subtilis and was thus proposed a potentially new way for producing recombinant SjLys protein.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Alma BP, Adriana TM, Aldo AA et al (2012) Shrimp invertebrate lysozyme i-lyz: Gene structure, molecular model and response of c and i lysozymes to lipopolysaccharide (LPS). Fish Shellfish Immun 32:230–236

    Article  Google Scholar 

  2. Avsaroglu MD, Buzrul S, Sanlibaba P et al (2007) A kinetic study on the plasmid stability of three Lactococcus lactis strains. J Ind Microbiol Biotechnol 34(11):729–737

    Article  CAS  Google Scholar 

  3. Bathige SD, Umasuthan N, Kasthuri SR et al (2013) A bifunctional invertebrate-type lysozyme from the disk abalone, Haliotis discus: Genome organization, transcriptional profiling and biological activities of recombinant protein. Dev Comp Immunol 41:282–294

    Article  CAS  Google Scholar 

  4. Chittibabu G, Ma C, Netter HJ et al (2014) Production, characterization, and immunogenicity of a secreted form of Plasmodium falciparum merozoite surface protein 4 produced in Bacillus subtilis. Appl Microbiol Biotechnol 98:3669–3678

    Article  CAS  Google Scholar 

  5. Cong LN, Ruan QK, Liu ZW et al (2013) Isolation of an antimicrobial marine strain HS-A38 and purification of its bioactive substances. J Pure Appl Microbio 7(1):379–385

    CAS  Google Scholar 

  6. Cong LN, Yang XJ, Wang XX et al (2009) Characterization of an i-type lysozyme gene from the sea cucumber Stichopus japonicus, and enzymatic and nonenzymatic antimicrobial activities of its recombinant protein. J Biosci Bioeng 107(6):583–588

    Article  CAS  Google Scholar 

  7. Dong XP, Zhu BW, Sun LM et al (2011) Changes of collagen in sea cucumber (Stichopus japonicas) during cooking. Food Sci. Biotechnol. 20(4):1137–1141

    Article  CAS  Google Scholar 

  8. Fatma MM, Carlos DD, Marc B (2012) High transformation efficiency of Bacillus subtilis with integrative DNA using glycine betaine as osmoprotectant. Anal Biochem 424:127–129

    Article  Google Scholar 

  9. Gao FY, Qu L, Yu SG et al (2012) Identification and expression analysis of three c-type lysozymes in Oreochromis aureus. Fish Shellfish Immun 32:779–788

    Article  CAS  Google Scholar 

  10. Leen DG, Jos MP, Paul J et al (2007) Stability of a promiscuous plasmid in different hosts: no guarantee for a long-term relationship. Microbiology 153:452–463

    Article  Google Scholar 

  11. Li SS, Wen JP, Jia XQ (2011) Engineering Bacillus subtilis for isobutanol production by heterologous Ehrlich pathway construction and the biosynthetic 2-ketoisovalerate precursor pathway overexpression. Appl Microbiol Biotechnol 91:577–589

    Article  CAS  Google Scholar 

  12. Liu HZ, Zheng FR, Sun XQ et al (2012) Construction of cDNA library from intestine, Mesentery and Coelomocyte of Apostichopus japonicus selenka infected with Vibrio sp. and a preliminary analysis of immunity-related genes. J. Ocean Univ. China 11(2):187–196

    Article  CAS  Google Scholar 

  13. Liu YH, Lin S, Zhang XQ et al (2014) A novel approach for improving the yield of Bacillus subtilis transglutaminase in heterologous strains. J Ind Microbiol Biotechnol 41:1227–1235

    Article  CAS  Google Scholar 

  14. Mu L, Wen JP (2013) Engineered Bacillus subtilis 168 produces L-malate by heterologous biosynthesis pathway construction and lactate dehydrogenase deletion. World J Microbiol Biotechnol 29:33–41

    Article  CAS  Google Scholar 

  15. Ranjita B, Masaru Y, Hideki N et al (2012) Enhanced production of 2,3-butanediol by engineered Bacillus subtilis. Appl Microbiol Biotechnol 94:651–658

    Article  Google Scholar 

  16. Reza P, Ebrahim VF, Seyed AS et al (2014) Induction of Bacillus subtilis expression system using environmental stresses and glucose starvation. Ann Microbiol l64:879−882

    Google Scholar 

  17. Shoham Y, Israeli E, Sonensheim AL et al (1991) Inhibition of growth of Bacillus subtilis by recombinant plasmid pCED3. Arch Microbiol 156:204–212

    Article  CAS  Google Scholar 

  18. Sun LM, Zhu BW, Wu HT et al (2011) Purification and characterization of Cathepsin B from the Gut of the Sea Cucumber (Stichopus japonicas). Food Sci Biotechnol 20(4):919–925

    Article  CAS  Google Scholar 

  19. Wang XY, Chen WJ, Tian YL et al (2014) Surface display of Clonorchis sinensis enolase on Bacillus subtilis spores potentializes an oral vaccine candidate. Vaccine 32:1338–1345

    Article  CAS  Google Scholar 

  20. Wu HT, Li DM, Zhu BW et al (2013) Characterization of acetylcholinesterase from the gut of sea cucumber Stichopus japonicus. Fish Sci 79:303–311

    Article  CAS  Google Scholar 

  21. Yang AF, Zhou ZC, Dong Y et al (2010) Expression of immune-related genes in embryos and larvae of sea cucumber Apostichopus japonicus. Fish Shellfish Immunol 29:839–845

    Article  CAS  Google Scholar 

  22. Yuan Y, Feng F, Chen L et al (2014) Surface display of Acetobacter pasteurianus AdhA on Bacillus subtilis spores to enhance ethanol tolerance for liquor industrial potential. Eur Food Res Technol 238:285–293

    Article  CAS  Google Scholar 

  23. Yue X, Wang HX, Huang XH et al (2012) Single nucleotide polymorphisms in i-type lysozyme gene and their correlation with vibrio-resistance and growth of clam Meretrix meretrix based on the selected resistance stocks. Fish Shellfish Immunol 33:559–568

    Article  CAS  Google Scholar 

  24. Zhang HW, Sun C, Sun SS et al (2010) Functional analysis of two invertebrate-type lysozymes from red swamp crayfish, Procambarus clarkia. Fish Shellfish Immunol 29:1066–1072

    Article  CAS  Google Scholar 

  25. Zou MY, Li XZ, Shi WJ et al (2013) Improved production of alkaline polygalacturonate lyase by homologous overexpression pelA in Bacillus subtilis. Process Biochem 48:1143–1150

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the Natural Science Foundation of China (Grant no. 31072224) and Natural Science Foundation of Liaoning Province of China (Grant no. 20102009).

Author information

Authors and Affiliations

  1. College of Bioengineering, Dalian Polytechnic University, Dalian, 116034, Liaoning, People’s Republic of China

    Zhiwen Liu, Xingyu Liao, Lu Sun, Dan Zou, Dan Li & Lina Cong

Authors
  1. Zhiwen Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xingyu Liao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lu Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dan Zou
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Lina Cong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lina Cong .

Editor information

Editors and Affiliations

  1. Tianjin University of Science and Technology, Tianjin, China

    Tong-Cun Zhang

  2. SBI ALApromo, Tokyo, Japan

    Motowo Nakajima

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer-Verlag Berlin Heidelberg

About this paper

Cite this paper

Liu, Z., Liao, X., Sun, L., Zou, D., Li, D., Cong, L. (2015). Expression of Stichopus japonicus Lysozyme Gene in Bacillus subtilis WB600. In: Zhang, TC., Nakajima, M. (eds) Advances in Applied Biotechnology. Lecture Notes in Electrical Engineering, vol 332. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-45657-6_15

Download citation

Publish with us

Policies and ethics

Search

Navigation