花生种子萌发过程中子叶内肽酶活性研究

Study on endopeptidase activity during peanut seeds germination

探讨花生种子在萌发过程中子叶内肽酶的活性及其同工酶数量的变化,研究金属型蛋白酶抑制剂EDTA与不同温度对内肽酶的活性的影响。采用直立玻板法萌发花生种子,运用分光光度法测定内肽酶的活力,结果发现内肽酶活力在种子萌发6 d达到最大值;运用SDS-PAGE凝胶电泳法发现在种子萌发6 d出现5条内肽酶同工酶(其中2条是金属型内肽酶);在种子萌发8 d和10 d均出现7条内肽酶同工酶(其中4条是金属型内肽酶),这些内肽酶同工酶的最适温度约为40℃,EGTA对萌发6 d时存在的2种金属型内肽酶同工酶有明显抑制作用,但并不能完全抑制其活性。

The activity and isozyme of endopeptidases during peanut seeds germination were detected to research the activity of endopeptidase affected by different temperatures and inhibitors of metalloprotease EDTA. In this study, we used the standing boards to germinate peanut seeds, used spectrophotometry to determine the activity of endopeptidase during the germination of seeds. It was found that the ativity of endopeptidase reached the maximum at the sixth day. Using SDS-PAGE gel electrophoresis, we found 5 kinds of endopeptidase isoenzymes, in which there were two kind of metalloprotease of the 6 days of germination, and 7 kinds of endopeptidase isoenzymes, which there were 4 kind of metalloprotease after 8 days and 10 days of germination. The optimum temperature of these endopeptidase isozymes was about 40℃. Gelatin electrophoresis results showed that EGTA had inhibitory effect on two isozymes of metal type at 6 days of germination, but it did not completely inhibit the activity.