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- Title
甘蓝型油菜Kunitz蛋白酶抑制剂的异源表达与理化特征分析.
- Authors
冯玮; 李朋朋; 宋鹏
- Abstract
A Kunitz inhibitor gene (RTI;rti)from rapeseed was cloned and expressed in Pichia pastoris expression system for the first time. Its physicochemical characteristics were investigated after separation and purification. The results showed that the expression level of recombinant RTI reached 628 mg·L-1 and its inhibitory activity reached 69.6 TIU·mg-1 in shaking flask fermentation. The recombinant RTI maintained over 70% inhibitory activity between 30-90 ℃ and retained over 80% inhibitory activity between pH 2.0-11.0. The metal ions Cu2+ and Co2+, the organic reagents methanol, ethanol, acetone or chloroform inhibited the activity of RTI. The recombinant RTI reacted with trypsin in a non-competitive manner, and demonstrated a strong and specific inhibitory effect on trypsin. These results indicated that RTI was a typical plant Kunitz trypsin inhibitor. Combined with good physicochemical properties, RTI has the potential to be developed as insect resistant protein.
- Publication
Acta Agriculturae Zhejiangensis, 2022, Vol 34, Issue 1, p112
- ISSN
1004-1524
- Publication type
Article
- DOI
10.3969/j.issn.1004-1524.2022.01.14