SDS-PAGE法测定蒜酶相对分子质量及含量

Determination of relative molecular weight and content of alliinase by SDS-PAGE method

目的采用还原型SDS-聚丙烯酰胺凝胶(SDS-PAGE)法测定蒜酶的相对分子质量及含量。方法建立测定蒜酶的凝胶电泳条件:分离胶12%,浓缩胶5%,初始电压80V,进入分离胶时电压调至100V,测定经提取纯化后的蒜酶和大蒜样品中蒜酶的分子质量及含量。结果通过SDS-PAGE法确定以磷酸盐缓冲液为溶剂,蒜酶供试品浓度27.2 mg/mL进行蒜酶含量和相对分子质量测定;蒜酶供试品中蒜酶平均相对分子质量50.84KDa,蒜酶供试品中蒜酶在总蛋白条带中平均相对含量为52.39%,大蒜中蒜酶在总蛋白条带中平均相对含量为6.28%。结论 SDS-PAGE法测定蒜酶含量及其相对分子质量,方法简便易行、结果准确可靠。

Objective The relative molecular mass and content of alliinase were determined by reducing SDS-polyacrylamide gel（SDS-PAGE）method.Methods The gel electrophoresis conditions for the determination of garlic enzyme were determined.The gel of separation was 12%,the gel of concentration was 5%,the initial voltage was 80 V,and the separation gel was adjusted to 100 V.The alliinase in the extracted garlic and garlic sample was determined.Results The SDS-PAGE method was used to determine the enzyme content and relative molecular mass of garlic with phosphate buffer solution as the solvent and the concentration of alliinase was 27.2 mg/mL.The average relative molecular mass of alliinase was 50.84 KDa,The relative content of alliinase in the total protein band of the enzyme test sample reached 52.39%.garlic was used as the sample,the relative content of garlic enzyme in the total protein band reached 6.28%.Conclusion The SDS-PAGE method was used to determine the alliinase content and molecular weight.The method was simple,and the result was accurate and reliable.