胞壁质酶的性质研究

目的]以提取的胞壁质酶为材料,研究胞壁质酶的性质。方法]通过测定胞壁质酶在595nm处光吸收值的增量,测定该酶活力和蛋白质浓度。通过测定米氏常数Km,研究酶促反应的速度及影响速度的因素。在不同pH值缓冲液中测定胞壁质酶活力,同时测定该酶的最适pH值。酶促反应的速度在酶的最适温度时达到最大。利用SDS-PAGE电泳法,测定胞壁质酶的分子量及纯度。结果]胞壁质酶在219.00g/L光吸收下降最快,活力最高。Km为0.57。脲对胞壁质酶为抑制作用,其抑制类型为竞争性抑制。胞壁质酶在pH值为8.0时酶活力最高,属于弱碱性,在40℃时酶活力最高,其Kd为13。结论]该研究可为今后采用生物工程技术对胞壁质酶进行克隆、提取以及制取提供参考。

Study of Properties of Cell Wall Lysozyme

Objective]The properties of cell wall lysozyme extracted were studied.Method] The enzyme activity and protein concentration were measured by measuring the increment of the absorption of lysozyme under 595 nm light.Michaelis constant Km was determined by examining the rate of enzymatic reaction speed and the impact of various factors.Enzyme activity of buffers under conditions of different pH was measured to determine the optimum enzyme pH.The optimal enzyme temperature could be determined by measuring the effects of different temperature on enzymatic reaction.The purity and molecular weight of lysozyme could be determined by the method of SDS-PAGE electrophore.Result] The light absorption value of lysozyme declined under the fastest speed in 219.00 g/L protein.The value of Michaelis constant Km was 0.57.Urea inhibited the quality of lysozyme which was the type of competitive inhibition.The most optimum enzyme pH for lysozyme was 8.0 which was weakly alkaline.Lysozyme had the highest enzyme activity under 40 ℃.Kd of lysozyme was 13.Conclusion] The research could provide reference of extraction of biological engineering technology.