苦瓜籽MAP30酶活性试验研究

Study on the Enzymatic Activity of MAP30 of Momordica Charantia

建立一种快速、高效检测MAP30酶活性的方法,并初步研究其部分生物活性。利用MAP30能以PET28a DNA为底物并脱去一些腺嘌呤碱基使超螺旋DNA解旋的特点,建立了测定MAP30酶活性的一种新方法,检测MAP30的酸碱和温度酶活性。MAP30灵敏度1μg,酶催化反应的时间是30 min。最适温度和p H值是37℃和8.0。不同p H值环境对MAP30特性的影响是不同的,酸性越强或碱性越强对MAP30的影响越大。不同温度值环境对MAP30特性的影响是不同的,温度越高MAP30活性损失。因此,合适的酸碱和温度为广泛和深入地研究MAP30酶活性提供了有利的条件。

To establish a rapid and efficient method for preparation of MAP30 enzymatic activity and to study preliminarily its biological activity,principle of the method is based on that MAP30 can remove some adenine bases from double stranded supercoiled PET28 a DNA molecules,subsequently,the deadenylated DNA was cleaved into nicked and linear form. The deadenylated DNA was degraded into many small fragments and run out of the gel. The optimum techniques of acid-base and temperature stability were studied,in order to further investigate enzyme characteristic and technique of resolution. The enzymatic activities of MAP30 were tested using this method. The limit of sensitivity is about 1 μg. The optimal p H and temperature of MAP30 was 8. 0 and 37 ℃,respectively.Different p H and temperature value conditions influence the characteristics of the MAP30 and,the higher acidity or alkalinity and temperature of the chemical solution,the more enzymatic activities influence on the intensity of the MAP30. Therefore,acidity or alkalinity is one of the main factors influencing the enzymatic activity of MAP30. It should be emphasized that the merit of this method is to avoid the preparation of ribosome.