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

Authors

SHAGHAYEGH NAMVAR
FARZANEH BARKHORDARI
MOZHGAN RAIGANI
HODA JAHANDAR
LEILA NEMATOLLAHI
FATEMEH DAVAMI

DOI

10.3906/biy-1708-12

Abstract

${\alpha}$-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.

Keywords

E. coli expression system, fed-batch culture, ribosome inactivating proteins, soluble expression, ${\alpha}$-luffin

First Page

23

Last Page

32

Recommended Citation

NAMVAR, SHAGHAYEGH; BARKHORDARI, FARZANEH; RAIGANI, MOZHGAN; JAHANDAR, HODA; NEMATOLLAHI, LEILA; and DAVAMI, FATEMEH (2018) "Cloning and soluble expression of mature ${\alpha}$-luffin from Luffa cylindrica in E. coli using SUMO fusion protein," Turkish Journal of Biology: Vol. 42: No. 1, Article 3. https://doi.org/10.3906/biy-1708-12
Available at: https://journals.tubitak.gov.tr/biology/vol42/iss1/3