Skip to main content
SpringerLink
Log in

Expression and secretion of a single-chain sweet protein, monellin, in Saccharomyces cerevisiae by an α-factor signal peptide

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

The sweet protein monellin gene was expressed in Saccharomyces cerevisiae under the control of the GAL1 promoter and α-factor signal peptide sequence of S. cerevisiae. The gene, which was obtained through mutation of the synthesized single-chain monellin gene, was cloned into an E. coli-yeast shuttle vector pYES2.0 which carries the galactose-inducible promoter GAL1. Then the α-factor signal peptide of S. cerevisiae was linked also, resulting in the secreting expression vector pYESMTA. The recombinant plasmid was subsequently transformed into strain S. cerevisiae INVsc1. The peptide efficiently directed the secretion of monellin from the recombinant yeast cell. A maximum yield of active monellin was 0.41 g l−1 of the supernatant from INVsc1 harboring pYESMTA.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Chen ZJ, Cai H, Lu FP, Du LX (2005) High-level expression of a synthetic gene encoding a sweet protein, monellin, in Escherichia coli. Biotechnol Lett 27:1745–1749

    Article  PubMed  CAS  Google Scholar 

  • Chen ZJ, Cai H, Li ZY, Liang XL, Shangguan XC (2007) Expression and secretion of a single-chain sweet protein monellin in Bacillus subtilis by sacB promoter and signal peptide. Appl Microbiol Biotechnol 73:1377–1381

    Article  PubMed  CAS  Google Scholar 

  • Douglas M, Maria JQ, Maria DL, Cruz MC, Sebastian C (2008) Systems for applied gene control in Saccharomyces cerevisiae. Biotechnol Lett 30:979–987

    Article  Google Scholar 

  • Gietz RD, Woods RA (2002) Transformation of yeast by the LiAc/ss carrier DNA/PEG method. Methods Enzymol 350:87–96

    Article  PubMed  CAS  Google Scholar 

  • Kim SH, Kang CH, Cho JM, Lee TK (1989) Redesigning a sweet protein: increased stability and renaturability. Protein Eng 2:571–575

    Article  PubMed  CAS  Google Scholar 

  • Kondo K, Miura Y, Song H, Kobayashi K (1997) High level expression of a sweet protein, monellin, in the food yeast Candida utilis. Nat Biotechnol 15:453–457

    Article  PubMed  CAS  Google Scholar 

  • Liu WF, Gao D, Bao XM (1998) The study of functional performance of inducing-promoter in S. cerevisiae. J Shandong Univ (Chin) 33:345–349

    Google Scholar 

  • Morris JA, Cagan RH (1972) Purification of monellin, the sweet principle of Discorephyllum cumminisii. Biochiem Biophys Acta 261:114–122

    CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Southgate VJ, Steyn AJ, Pretorius IS, Van Vuuren HJ (1993) Expression and secretion of Bacillus amyloliquefaciens alpha-amylase by using the yeast pheromone alpha-factor promoter and leader sequence in Saccharomyces cerevisiae. Appl Environ Microbiol 59:1253–1258

    PubMed  CAS  Google Scholar 

  • Zhang XL, Ito T, Kondo K, Kobayashi T, Honda H (2002) Production of single chain recombinant monellin by high cell density culture of genetically engineered Candida utilis using limited feeding of sodium ions. J Chem Eng Jpn 35:654–659

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. College of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot, 010018, People’s Republic of China

    Zhongjun Chen, Zhengying Li, Na Yu & Lili Yan

Authors
  1. Zhongjun Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhengying Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Na Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lili Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhongjun Chen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chen, Z., Li, Z., Yu, N. et al. Expression and secretion of a single-chain sweet protein, monellin, in Saccharomyces cerevisiae by an α-factor signal peptide. Biotechnol Lett 33, 721–725 (2011). https://doi.org/10.1007/s10529-010-0479-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-010-0479-2

Keywords

Search

Navigation