Cloning and soluble expression of mature ${\alpha}$-luffin from Luffa cylindrica in E. coli using SUMO fusion protein

Turkish Journal of Biology

SHAGHAYEGH NAMVAR¹ 

关键词: E. coli expression system;    fed-batch culture;    ribosome inactivating proteins;    soluble expression;    ${\alpha}$-luffin;   

学科分类：生物科学（综合）

来源: Scientific and Technical Research Council of Turkey - TUBITAK

【 摘 要 】

${\alpha}$-Luffin, found in Luffa cylindrica seeds, is a type I ribosome inactivating proteins. Cytotoxic effects make it an appropriate candidate for the construction of immunotoxins and conjugates. Because of limited natural resources, recombinant technology is the best approach to achieve large-scale production of plant-based proteins. In the present study, ${\alpha}$-luffin protein was expressed in E. coli and the effects of different temperature conditions, SUMO fusion tag, and cultivation strategies on total expression and solubility were investigated. Protein expression was evaluated at different intervals (0, 4, 6, 8, 24 h) postinduction. Our results showed that EnBase had higher efficiency than LB, and maximum solubility and total protein expression were achieved 24 h after induction at 30 °C and 25 °C, respectively. It was shown that SUMO tag is an effective strategy to improve protein solubility.

【 授权许可】

Unknown