Namvar Shaghayegh, Barkhordari Farzaneh, Raigani Mozhgan, Jahandar Hoda, Nematollahi Leila, Davami Fatemeh
Biotechnology Research Center; Pasteur Institute of Iran , Tehran.
Pharmaceutical Sciences Research Center, Pharmaceutical Sciences Branch, Islamic Azad University , Tehran , Iran.
Turk J Biol. 2018 Feb 15;42(1):23-32. doi: 10.3906/biy-1708-12. eCollection 2018.
α-Lufin, found in Luaf 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, α-lufin 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 eficiency 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.
α-卢芬存在于圆柱叶卢菲种子中，是一种I型核糖体失活蛋白。细胞毒性作用使其成为构建免疫毒素和缀合物的合适候选物。由于自然资源有限，重组技术是实现大规模生产植物源蛋白的最佳方法。在本研究中，α-卢芬蛋白在大肠杆菌中表达，并研究了不同温度条件、SUMO融合标签和培养策略对总表达量和溶解度的影响。在诱导后的不同时间间隔（0、4、6、8、24小时）评估蛋白表达情况。我们的结果表明，EnBase比LB具有更高的效率，在30°C和25°C诱导24小时后分别实现了最大溶解度和总蛋白表达量。结果表明，SUMO标签是提高蛋白溶解度的有效策略。
