標題: Enhancing the stability of xylanase from Cellulomonas fimi by cell-surface display on Escherichia coli
作者: Chen, Y-P
Hwang, I-E
Lin, C-J
Wang, H-J
Tseng, C-P
生物科技學系
Department of Biological Science and Technology
關鍵字: anchor protein;cell-surface display;thermostability;xylanase
公開日期: 1-三月-2012
摘要: 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 49 U g) 1 dry cell weight after 12 h culture at 37 degrees 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 degrees 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.
URI: http://dx.doi.org/10.1111/j.1365-2672.2012.05232.x
http://hdl.handle.net/11536/15613
ISSN: 1364-5072
DOI: 10.1111/j.1365-2672.2012.05232.x
期刊: JOURNAL OF APPLIED MICROBIOLOGY
Volume: 112
Issue: 3
起始頁: 455
結束頁: 463
顯示於類別:期刊論文


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