超声波和加热处理对三疣梭子蟹肝胰腺丝氨酸蛋白酶的影响

Effects of Ultrasound and Heat Treatments on the Activity of Serine Protease in Hepatopancreas from Swimming Crab(Portunus trituberculatus)

以三疣梭子蟹(Portunus trituberculatus)肝胰腺中丝氨酸蛋白酶为研究对象,对其进行超声波和加热处理,通过紫外光谱、荧光光谱分析和蛋白质变性凝胶电泳分析丝氨酸蛋白酶的变化。研究表明,超声功率和超声时间对丝氨酸蛋白酶的酶活有不同影响。低功率短时间超声处理会使酶活力升高,随着功率和超声时间的增加,酶活力逐渐降低。紫外光谱显示,超声处理使酶液紫外吸收出现蓝移现象,而加热对其影响不大。内源荧光显示,超声和加热处理均对蛋白酶有荧光猝灭作用。SDS-PAGE显示,超声和加热2种处理方法均能引起丝氨酸蛋白酶不同程度的降解。

Ultraviolet and fluorescence spectroscopies and SDS-PAGE were used to determine the change of serine protease activity in swimming crab hepatopancreas after ultrasound and heating treatment.The results showed that ultrasonic power and ultrasonic time had different impacts on the activity of serine protease.Low ultrasonic power and short ultrasonic time could activate enzyme activity,however when the ultrasonic power and time increased,the activity reduced gradually.Ultraviolet spectroscopy showed that ultraviolet absorption of serine protease after ultrasonic treatment appeared blue-shifted phenomenon and there was no difference on ultraviolet absorption of the enzyme after heat treatment.Fluorescence spectroscopy showed that the two treatments resulted in a fluorescence quenching phenomenon.SDS-PAGE showed that both ultrasonic and heat treatments can degrade the enzyme to various degrees.