Enhanced expression of cysteine-rich antimicrobial peptide snakin-1 in Escherichia coli using an aggregation-prone protein coexpression system

Authors

  • Md. Ruhul Kuddus,

    1. Graduate School of Life Science, Hokkaido University, Sapporo, Hokkaido, Japan
    2. Dept. of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Dhaka, Dhaka, Bangladesh
    Search for more papers by this author
    • Md. Ruhul Kuddus and Megumi Yamano contributed equally to this study

  • Megumi Yamano,

    1. Graduate School of Life Science, Hokkaido University, Sapporo, Hokkaido, Japan
    Search for more papers by this author
    • Md. Ruhul Kuddus and Megumi Yamano contributed equally to this study

  • Farhana Rumi,

    1. Graduate School of Life Science, Hokkaido University, Sapporo, Hokkaido, Japan
    Search for more papers by this author
  • Takashi Kikukawa,

    1. Graduate School of Life Science, Hokkaido University, Sapporo, Hokkaido, Japan
    2. Global Station for Soft Matter, Global Inst. for Collaborative Research and Education, Hokkaido University, Sapporo, Japan
    Search for more papers by this author
  • Makoto Demura,

    1. Graduate School of Life Science, Hokkaido University, Sapporo, Hokkaido, Japan
    2. Global Station for Soft Matter, Global Inst. for Collaborative Research and Education, Hokkaido University, Sapporo, Japan
    Search for more papers by this author
  • Tomoyasu Aizawa

    Corresponding author
    1. Graduate School of Life Science, Hokkaido University, Sapporo, Hokkaido, Japan
    2. Global Station for Soft Matter, Global Inst. for Collaborative Research and Education, Hokkaido University, Sapporo, Japan
    Search for more papers by this author

Abstract

Snakin-1 (SN-1) is a cysteine-rich plant antimicrobial peptide and the first purified member of the snakin family. SN-1 shows potent activity against a wide range of microorganisms, and thus has great biotechnological potential as an antimicrobial agent. Here, we produced recombinant SN-1 in Escherichia coli by a previously developed coexpression method using an aggregation-prone partner protein. Our goal was to increase the productivity of SN-1 via the enhanced formation of insoluble inclusion bodies in E. coli cells. The yield of SN-1 by the coexpression method was better than that by direct expression in E. coli cells. After refolding and purification, we obtained several milligrams of functionally active SN-1, the identity of which was verified by MALDI-TOF MS and NMR studies. The purified recombinant SN-1 showed effective antimicrobial activity against test organisms. Our studies indicate that the coexpression method using an aggregation-prone partner protein can serve as a suitable expression system for the efficient production of functionally active SN-1. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1520–1528, 2017

Ancillary