Heterologous expression and characterization of the hydrophobin HFBI in Pichia pastoris and evaluation of its contribution to the food industry
Heterologous expression and characterization of the hydrophobin HFBI in Pichia pastoris and evaluation of its contribution to the food industry
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毕赤酵母中疏水蛋白HFBI的异源表达和表征及其对食品工业的贡献评估
DOI：
10.1007/s00726-011-1126-5
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发表时间：
2011-10
期刊：
影响因子：
3.5
通讯作者：
Qiao, Mingqiang
中科院分区：
文献类型：
--
作者：
Wang, D;an;Yang, Yanyan;Xu, Haijin;Qiao, Mingqiang
利用pPIC 9载体，在AOX 1启动子的调控下，在毕赤酵母中异源表达里氏木霉II类疏水蛋白HFBI。重组HFBI（rHFBI）经超滤和反相高效液相色谱纯化。Tricine-SDS-PAGE和Western blotting结果表明，重组HFBI可分泌到培养液中，分子量约为7.5kDa。X射线光电子能谱和水接触角的测量表明，rHFBI可以导致依赖于疏水/亲水界面上的自组装膜的疏水硅化玻璃和亲水云母表面的润湿性的转换。结果表明，rHFBI具有稳定油滴的能力，其稳定油滴的能力远远超过在巴斯德毕赤酵母中异源表达的I类疏水蛋白HGFI（rHGFI）和典型的食品乳化剂酪蛋白酸钠。在喷涌实验中，它表明，rHFBI是一个强大的喷涌诱导剂在啤酒中，而rHGFI没有显示任何喷涌的迹象。这提供了潜在的rHFBI被用作一种新的乳化剂和预测喷涌的风险。
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.
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影响因子：
10.5
作者：
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通讯作者：
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影响因子：
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作者：
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作者：
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