Enhancing the stability of xylanase from Cellulomonas fimi by cell-surface display on Escherichia coli

Y. P. Chen, I. E. Hwang, C. J. Lin, H. J. Wang, Ching-Ping Tseng*

*Corresponding author for this work

Research output: Contribution to journalArticle

22 Scopus citations

Abstract

Aims: The cell-surface display of Cex, which encodes xylanase and exoglucanase from Cellulomonas fimi, was constructed on Escherichia coli using PgsA as the anchor protein. Characterization of the cell-surface display of Cex was performed. Methods and Results: PgsA was fused to the N-terminus of Cex and six histidines were utilized as spacers between the targeting and anchor proteins. Successful cell-surface display of Cex was demonstrated by Western blot and immunofluorescence analyses on E. coli C41 (DE3). According to the time-course analysis, the xylanase activity of Cex was achieved at 49Ug -1 dry cell weight after 12h culture at 37°C. The optimal temperature and pH ranges of the cell-surface displayed protein with whole-cell were broader than the corresponding ranges of the purified form. Further determination of thermostability indicated that the half-life of cell-surface displayed Cex was 1·6 times longer than that of purified Cex at 60°C. Conclusions: We have successfully developed the cell-surface display of xylanase on E. coli. The cell-surface display can enhance the stability of xylanase against changes in temperature and has the potential of becoming a whole-cell biocatalyst for industrial applications, such as biobleaching of paper and production of renewable energy. Significance and Impact of the Study: The results demonstrated that the cell-surface display of xylanase embedded in the cell membrane is more stable than that of the purified enzyme. Thus, to improve the stability of heterologous proteins production, cell-surface display using the PgsA anchor protein as a tool can be considered in E. coli.

Original languageEnglish
Pages (from-to)455-463
Number of pages9
JournalJournal of Applied Microbiology
Volume112
Issue number3
DOIs
StatePublished - 1 Mar 2012

Keywords

  • Anchor protein
  • Cell-surface display
  • Thermostability
  • Xylanase

Fingerprint Dive into the research topics of 'Enhancing the stability of xylanase from Cellulomonas fimi by cell-surface display on Escherichia coli'. Together they form a unique fingerprint.

  • Cite this