Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/60332
標題: A novel lysozyme from Xanthomonas oryzae phage phi Xo411 active against Xanthomonas and Stenotrophomonas
作者: Lee, C.N.
翁淑芬
Lin, J.W.
Chow, T.Y.
Tseng, Y.H.
Weng, S.F.
關鍵字: Xanthomonas
Stenotrophomonas
bacteriophage
lysozyme
escherichia-coli
bacteriophage-lambda
t4 lysozyme
egg-white
purification
gene
endolysin
pathogens
virulence
release
期刊/報告no:: Protein Expression and Purification, Volume 50, Issue 2, Page(s) 229-237.
摘要: In this study, a bacteriophage of Xanthomonas oryzae pv. oryzae designated as phi Xo411 was isolated. Random sequencing of its genome revealed that it is closely related to another X. oryzae phage, Xp10. A cloned fragment carries the lysozyme gene, lys411. The deduced protein, Lys411, shares 92% identity with Xp10 lysozyme, which contains an extra 46 aa at the N-terminus. Lys411 shows over 40% identities to several other phage lysozymes. The His-tagged protein, Lys411H, expressed in Escherichia coli largely formed as inclusion bodies. The insoluble protein was solubilized in urea and purified by passing through a His-bind column, and the lytic activity was then restored by a refolding process. The optimal assay conditions determined for Lys411H are in 0.1 M potassium phosphate buffer, pH 6.6 containing 1 mM CuCl2 at 25 degrees C. Lysis assays using different bacterial cells as the substrates indicate that Lys411H is the first lysozyme active against both Xanthomonas and Stenotrophomonas maltophilia. This suggests that Lys411 can be a candidate to be developed into a therapeutic agent for treating S. maltophilia infections, in addition to the potential use in control of the plant diseases caused by Xanthomonas. By analogy to the situation in Xp10, we predict that phi Xo411 has no holin, the protein required for lysozyme export, and the N-terminal signal-arrest-release sequence of Lys411 can accommodate its own export to the periplasm. (c) 2006 Elsevier Inc. All rights reserved.
URI: http://hdl.handle.net/11455/60332
ISSN: 1046-5928
文章連結: http://dx.doi.org/10.1016/j.pep.2006.06.013
Appears in Collections:分子生物學研究所

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