Using Rice Flour for Purification of Maltose Binding Fusion Proteins Expressed in Escherichia coli

authors:

avatar Azadeh Neissi 1 , avatar Masoudreza Seyfi Abad Shapouri 1 , * , avatar Masoud Ghorbanpoor Najafabadi 1 , avatar Amin Jaydary 2

Department of Pathobiology, School of Veterinary Medicine, Shahid Chamran University, Ahvaz, IR Iran
Department of Pathobiology, School of Veterinary Medicine, University of Lorestan, Khorram Abad, IR Iran

how to cite: Neissi A, Seyfi Abad Shapouri M, Ghorbanpoor Najafabadi M, Jaydary A. Using Rice Flour for Purification of Maltose Binding Fusion Proteins Expressed in Escherichia coli. Jundishapur J Microbiol. 2013;6(3): 233-6. https://doi.org/10.5812/jjm.4926.

Abstract

Background:

One disadvantage of expressing heterologous proteins in Escherichia coli is that the proteins are frequently expressed as insoluble inclusion bodies. To avoid this problem, heterologous proteins are typically expressed as a fusion protein. Maltose binding protein (MBP) is one of the widely used partners for production of recombinant fusion proteins in E. coli. MBP is among the most effective solubility enhancers. In addition, MBP can be used as an affinity tag for purification of recombinant proteins on a column of amylose resin.

Objectives:

The purpose of this study was to investigate the use of rice flour, a natural source of amylose, for purification of MBP fusion proteins.

Materials and Methods:

MBP and a fusion protein of MBP and avian influenza virus nucleoprotein (MBP-NP) were expressed in E. coli and subjected to purification by rice flour and a commercial amylose resin. The purified proteins were analyzed by SDS-PAGE.

Results:

The results indicated that MBP and MBP-NP, both were successfully purified by rice flour.

Conclusions:

Rice flour can be used for purification of MBP fusion proteins. Although the efficiency of purification by rice flour is less than amylose resin, however, the yield is sufficient to obtain a quantity of protein required for research purposes.

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