Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewA

Int J Mol Sci. 2021 Jul 22;22(15):7843. doi: 10.3390/ijms22157843.

Abstract

Hydrophobins are small proteins (<20 kDa) with an amphipathic tertiary structure that are secreted by various filamentous fungi. Their amphipathic properties provide surfactant-like activity, leading to the formation of robust amphipathic layers at hydrophilic-hydrophobic interfaces, which make them useful for a wide variety of industrial fields spanning protein immobilization to surface functionalization. However, the industrial use of recombinant hydrophobins has been hampered due to low yield from inclusion bodies owing to the complicated process, including an auxiliary refolding step. Herein, we report the soluble expression of a recombinant class I hydrophobin DewA originating from Aspergillus nidulans, and its efficient purification from recombinant Escherichia coli. Soluble expression of the recombinant hydrophobin DewA was achieved by a tagging strategy using a systematically designed expression tag (ramp tag) that was fused to the N-terminus of DewA lacking the innate signal sequence. Highly expressed recombinant hydrophobin DewA in a soluble form was efficiently purified by a modified aqueous two-phase separation technique using isopropyl alcohol. Our approach for expression and purification of the recombinant hydrophobin DewA in E. coli shed light on the industrial production of hydrophobins from prokaryotic hosts.

Keywords: aqueous two-phase separation (ATPS); isopropyl alcohol (IPA); ramp tag (RT); recombinant hydrophobin DewA; soluble expression.

MeSH terms

  • Aspergillus nidulans / growth & development
  • Aspergillus nidulans / metabolism*
  • Fungal Proteins / genetics
  • Fungal Proteins / isolation & purification*
  • Fungal Proteins / metabolism*
  • Hydrophobic and Hydrophilic Interactions
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification*
  • Recombinant Proteins / metabolism*
  • Surface Properties

Substances

  • DEWA protein, Asperigillus nidulans
  • Fungal Proteins
  • Recombinant Proteins