Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/41156
標題: A comparative study on the secretion of alkaline phosphatase in Escherichia coli
作者: Huang, K.C.
林松池
Huang, P.H.
Lin, S.C.
關鍵字: Translocation
Secretion
Twin-arginine
Tat
Alkaline phosphatase
TorD
arginine translocation pathway
tat pathway
protein-transport
export
pathway
expression
complex
purification
membrane
bacteria
genes
期刊/報告no:: Journal of the Taiwan Institute of Chemical Engineers, Volume 40, Issue 1, Page(s) 29-35.
摘要: Secretion of alkaline phosphatase (AP), a homodimeric and disulfide bond-containing protein, fused with the YfhG and the TorA signal peptides via the Sec and the Tat pathways, respectively, in Escherichia coli were investigated. The efficiency of AP translocation via the Sec pathway in MC4100, 75.5 +/- 0.7%, was higher than that via the Tat pathway, 66.5 +/- 0.7%. The amount of active, periplasmic AP, as suggested by the results of AP activity analysis, translocated via the Sec pathway was more than threefold that via the Tat pathway. The efficient secretion of AP via the Sec pathway alleviated protein aggregations, thus reducing the formation of alkaline phosphatase inclusion bodies in the cytoplasm. The translocation of AP in E. coli strain DR473 with relatively oxidative cytoplasm confirms the dual specificity of the YfhG signal peptide for the Sec and Tat machineries. Secretion of AP via the Tat pathway in MC4100, on the other hand, suggests that the formation of disulfide bonds may not be necessary for AP secretion via the Tat pathway. The failure of co-expressing TorD, a molecular chaperone, in enhancing the translocation of AP via the Tat pathway in DR473, and the results of in vitro on-column binding experiments suggest that proper binding to the signal peptide by TorD is a prerequisite for enhancement in protein translocation. (c) 2008 Taiwan Institute of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
URI: http://hdl.handle.net/11455/41156
ISSN: 1876-1070
文章連結: http://dx.doi.org/10.1016/j.jtice.2008.06.002
Appears in Collections:化學工程學系所

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