Purification and Characterization of Elicitor-induced Chitinase from Grape Berries
Purification and Characterization of Elicitor-induced Chitinase from Grape Berries
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葡萄浆果中诱导子诱导的几丁质酶的纯化和表征
DOI：
10.5458/jag.47.163
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发表时间：
2000
影响因子：
1.1
通讯作者：
T. Takayanagi
中科院分区：
文献类型：
--
作者：
T. Uchibori;K. Yokotsuka;T. Takayanagi
从用甘醇几丁质诱导的甲州葡萄中纯化了诱导几丁质酶。通过使用 Chitopearl BL-3 和 Q-Sepharose HP (HiTrap Q) 柱进行连续色谱分离，从葡萄提取液中用硫酸铵盐析（80% 饱和度）获得蛋白质级分。诱导的几丁质酶在聚丙烯酰胺凝胶电泳上是均质的。诱导的几丁质酶的最适pH为4.0，并且该酶在pH 5.0-6.5之间的酸性条件下稳定。最适温度为40℃，酶在40℃以下稳定。诱导的几丁质酶的N端氨基酸序列为NH2-GTITVYXGQNGN，与葡萄III类几丁质酶高度同源。诱导的几丁质酶抑制灰霉病的生长，从而引起灰霉病，强烈表明它在成熟葡萄对抗植物病原体的防御机制中发挥着重要作用。
An induced chitinase was purified from Koshu grapes elicitated with glycolchitin. The protein fraction obtained by salting out with ammonium sulfate (80% saturation) from the extract solution of the grapes was separated by successive chromatographies on Chitopearl BL-3 and Q-Sepharose HP (HiTrap Q) columns. The induced chitinase was homogeneous on polyacrylamide gel electrophoresis. The optimum pH of the induced chitinase was 4.0, and the enzyme was stable under acidic conditions between pH 5.0-6.5. The optimum temperature was 40°C, and the enzyme was stable below 40°C. The N-terminal amino acid sequence of the induced chitinase was NH2-GTITVYXGQNGN, which is highly homologous with that of the class III chitinase of Vitis vinifera. The induced chitinase inhibited the growth of Botrytis cinerea, which causes grey mold disease, strongly suggesting that it plays an important role in the defense mechanism against phytopa-thogens in mature grape.