High-yield fermentation and a novel heat-precipitation purification method for hydrophobin HGFI from Grifola frondosa in Pichia pastoris

Dongmin Song, Zhendong Gao, Liqiang Zhao, Xiangxiang Wang, Haijin Xu, Yanling Bai, Xiuming Zhang, Markus B. Linder, Hui Feng, Mingqiang Qiao*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

10 Citations (Scopus)

Abstract

Hydrophobins are proteins produced by filamentous fungi with high natural-surfactant activities and that can self-assemble in interfaces of air–water or solid–water to form amphiphilic membranes. Here, we reported a high-yield fermentation method for hydrophobin HGFI from Grifola frondosa in Pichia pastoris, attaining production of 300 mg/L by keeping the dissolved oxygen level at 15%–25% by turning the methanol-feeding speed. We also developed a novel HGFI-purification method enabling large-scare purification of HGFI, with >90% recovery. Additionally, we observed that hydrophobin HGFI in fermentation broth precipitated at pH < 7.0 and temperatures >90 °C. We also identified the structure and properties of proteins purified by this method through atomic force microscopy, circular dichroism, X-ray photoelectron spectroscopy, and water-contact angle measurement, which is similar to protein purification by ultrafiltration without heating treatment that enables our method to maintain native HGFI structure and properties. Furthermore, the purification method presented here can be applied to large-scale purification of other type I hydrophobins.

Original languageEnglish
Pages (from-to)22-28
Number of pages7
JournalPROTEIN EXPRESSION AND PURIFICATION
Volume128
DOIs
Publication statusPublished - 1 Dec 2016
MoE publication typeA1 Journal article-refereed

Keywords

  • Characterization
  • Fed-batch fermentation
  • Hydrophobin HGFI
  • Purification

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