Novel Osmotically Induced Antifungal Chitinases and Bacterial Expression of an Active Recombinant Isoform
Novel Osmotically Induced Antifungal Chitinases and Bacterial Expression of an Active Recombinant Isoform
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DOI：
10.1104/pp.111.4.1219
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发表时间：
1996-08
期刊：
影响因子：
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通讯作者：
Dae-Jin Yun;M. P. d’Urzo;Laura R. Abad;S. Takeda;R. Salzman;Z. Chen;Hyeseung Lee;P. Hasegawa;R. Bressan
中科院分区：
文献类型：
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作者：
Dae-Jin Yun;M. P. d’Urzo;Laura R. Abad;S. Takeda;R. Salzman;Z. Chen;Hyeseung Lee;P. Hasegawa;R. Bressan
NaCl (428 mM)-adapted tobacco (Nicotiana tabacum L. var Wisconsin 38) cells accumulate and secrete several antifungal chitinases. The predominant protein secreted to the culture medium was a 29-kD peptide that, based on internal amino acid sequence, was determined to be a class II acidic chitinase with similarity to PR-Q. The four predominant chitinases (T1, T2, T3, and T4) that accumulated intracellularly in 428 mM NaCl-adapted cells were purified. Based on N-terminal sequence analyses, two of these were identified as class I chitinase isoforms, one similar to the N. tomentosiformis (H. Shinshi, J.M. Neuhaus, J. Ryals, F. Meins [1990] Plant Mol Biol 14: 357–368) protein (T1) and the other homologous to the N. sylvestris (Y. Fukuda, M. Ohme, H. Shinshi [1991] Plant Mol Biol 16: 1–10) protein (T2). The other two proteins (T3 and T4) were determined to be novel chitinases that have sequence similarity with class I chitinases, but each lacks a chitin-binding domain. All four chitinases inhibited Fusarium oxysporum f. sp. lycopersici and Trichoderma longibrachiatum hyphal growth in vitro, although the isoforms containing a chitin-binding domain were somewhat more active. Conditions were established for the successful expression of soluble and active bacterial recombinant T2. Expression of soluble recombinant T2 was achieved when isopropyl [beta]-D-thiogalactopyranoside induction occurred at 18[deg]C but not at 25 or 37[deg]C. The purified recombinant protein exhibited antifungal activity comparable to a class I chitinase purified from NaCl-adapted tobacco cells.
