乙醇对角质酶性质的影响

Effect of Ethanol on the Property of Cutinase

DOI：10.3969/j.issn.1673-1689.2012.01.005

中文关键词: 角质酶 乙醇 酶学性质 构象

英文关键词: cutinase ethanol enzymatic property conformation

基金项目:国家自然科学基金项目(20776063);国家计划863项目(2011AA100905)

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中文摘要:

研究了乙醇溶液对角质酶性质的影响,同时用圆二色光谱和荧光光谱法对经过硫酸铵沉淀、镍柱亲和层析等方法获得的电泳纯角质酶进行了构象研究。结果表明,在体积分数40%、30%和20%乙醇溶液中角质酶催化反应的最适温度分别为50、55和65℃;在37℃下,酶在体积分数20%～50%乙醇中具有较好的稳定性,体积分数30%和40%乙醇对角质酶具有激活作用;角质酶在体积分数30%和50%乙醇中催化反应的最适pH分别为9.5和9.0;在体积分数30%乙醇溶液中角质酶的pH稳定范围为6.5～10.5。角质酶在50℃的乙醇溶液中的失活动力学研究表明,角质酶的失活属于一级动力学反应(R2>0.98)。从角质酶构象的研究结果可以看出,在乙醇体系中角质酶的二级结构和三级结构都发生了相应的变化。

英文摘要:

In this manuscript,effect of ethanol solutions on the biochemical property and conformation of cutinase were studied.The results showed that the optimum temperatures of cutinase were 65 ℃,55 ℃ and 50 ℃ respectively in 20%(V/V),30% and 40% ethanol solutions.In 20% and 50% ethanol solutions cutinase was stable at 37℃.In 30% and 50% ethanol solutions the optimum pH of cutinase were 9.5 and 9.0 respectively.In 30% ethanol solution the pH stability range of cutinase was 6.5-10.5.In 30% and 40% ethanol solutions activity of cutinase at 37 ℃was enhanced.The inactivation kinetic of cutinase was researched in ethanol solutions at 50 ℃.The results showed that the inactivation kinetic of cutinase in ethanol solutions followed the first order reaction(R2>0.98).And the secondary and the tertiary structure conformations greatly changed in ethanol solutiods.

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