Amino Acids

, Volume 43, Issue 2, pp 763–771 | Cite as

Heterologous expression and characterization of the hydrophobin HFBI in Pichia pastoris and evaluation of its contribution to the food industry

  • Baolong Niu
  • Dandan Wang
  • Yanyan Yang
  • Haijin Xu
  • Mingqiang QiaoEmail author
Original Article


The class II hydrophobin HFBI from Trichoderma reesei was heterologously expressed by Pichia pastoris using pPIC9 vector under the control of the promoter AOX1. The recombinant HFBI (rHFBI) was purified by ultrafiltration and reverse-phase high performance liquid chromatography. Tricine-SDS-PAGE and Western blotting demonstrated that rHFBI with the expected molecular weight of 7.5 kDa was secreted into the culture medium. X-ray photoelectron spectroscopy and water contact angle measurements indicated that rHFBI could lead to the conversion of the wettability of the hydrophobic siliconized glass and hydrophilic mica surfaces relying on the self-assembly membrane on hydrophobic/hydrophilic interfaces. It was demonstrated that rHFBI had the ability to stabilize oil droplets, which was far excess of the class I hydrophobin HGFI heterologously expressed in P. pastoris (rHGFI) and the typical food emulsifier sodium caseinate. In gushing experiments, it was shown that rHFBI was a strong gushing inducer in beer, whereas rHGFI did not display any signs of gushing. This provided the potential of rHFBI to be used as a novel emulsifying agent and a predictor of gushing risk.


Hydrophobin Self-assembly Heterologous expression Emulsion Gushing 



This research was financially supported by the National Natural Science Foundation of China (Grant #31170066), 973 Program of China (Grant #2011CBA00802), Program for New Century Excellent Talents in University from the Ministry of Education of China (Grant #NCET-06-0212) and Sino-Finnish Scientific and Technological Cooperation Project from the Ministry of Science and Technology of China (Grant #2006DFA32360).

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Baolong Niu
    • 1
  • Dandan Wang
    • 1
  • Yanyan Yang
    • 1
  • Haijin Xu
    • 1
  • Mingqiang Qiao
    • 1
    Email author
  1. 1.The Key Laboratory of Bioactive MaterialsMinistry of Education, College of Life Sciences, Nankai UniversityTianjinPeople’s Republic of China

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