In the present study, α-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. | Turkish Journal of Biology Turk J Biol (2018) 42: 23-32 © TÜBİTAK doi: Research Article Cloning and soluble expression of mature α-luffin from Luffa cylindrica in E. coli using SUMO fusion protein 1,2, 2, 2 Shaghayegh NAMVAR *, Farzaneh BARKHORDARI *, Mozhgan RAIGANI , 1,3 2 2, Hoda JAHANDAR , Leila NEMATOLLAHI , Fatemeh DAVAMI ** 1 Pharmaceutical Sciences Research Center, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran 2 Biotechnology Research Center; Pasteur Institute of Iran, Tehran 3 Department of Biotechnology, Faculty of Advanced Sciences &Technology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran Received: Accepted/Published Online: Final Version: Abstract: α-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, α-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. Key words: E. coli expression system, fed-batch culture, ribosome inactivating proteins, soluble expression, α-luffin 1. Introduction α-Luffin is a single-chain type I ribosome-inactivating protein (RIP) found in Luffa cylindica seeds (Griffiths and Electricwala, 1987; Ng et
