Functional display of Rhizomucor miehei lipase on surface of Saccharomyces cerevisiae with higher activity and its practical properties

Functional display of Rhizomucor miehei lipase on surface of Saccharomyces cerevisiae with higher activity and its practical properties
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米赫根毛霉脂肪酶在酿酒酵母表面的高活性功能展示及其实用特性

DOI:
10.1002/jctb.1814
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发表时间:
2008-03
影响因子:
3.4
通讯作者:
Wei, Dong-Zhi
Wei, Dong-Zhi
中科院分区:
工程技术4区
文献类型:
--
作者:
Zhang, Wei-Guo;Wang, Xiao-Ning;Lin, Ying;Han, Shuang-Yan;Wei, Dong-Zhi

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背景:一种能够将DNA翻译成功能性多肽或蛋白质的展示系统被用作一种新的蛋白质表达系统。在这个系统中,多肽或蛋白质以与一些锚定蛋白融合的形式显示在细胞表面。酵母细胞表面有脂肪酶,可用作全细胞生物催化剂。本研究对米氏根霉脂肪酶(RML)在活性较高的酿酒酵母表面的功能展示进行了研究。 结果:米氏酵母3.4960产的脂肪酶为活性脂肪酶。表达的rm1-α-凝集素融合蛋白不分泌到培养上清液中,大部分固定在酵母细胞上。细胞表面脂肪酶在45℃、pH 8.0时酶活最高。 结论:利用糖基磷脂酰肌醇锚点可在酿酒酵母MT8-1细胞表面高效表达米氏酵母3.4960的rm1基因。版权所有(C)2007化学工业学会
BACKGROUND: A display system, which can translate DNA to functional peptides or proteins, is used as a new protein expression system. In this system, peptides or proteins are displayed on the cell surface as a fusion form with some anchoring proteins. Yeast cells displaying lipases on their cell-surface could be used as whole-cell biocatalysts. This research focuses on the functional display of Rhizomucor miehei lipase (RML) on the surface of Saccharomyces cerevisiae with higher activity. RESULTS: The lipases (RML) from R.miehei 3.4960 were of active form. The RML-α-agglutinin fusion proteins produced were not secreted into the culture media and were mostly immobilized on the yeast cells. Cell surface displayed lipase showed the highest activity at 45 °C and pH 8.0. CONCLUSION: The gene encoding RML from R.miehei 3.4960 can be functionally expressed on the cell surface of S. cerevisiae MT8-1 using a glycosylphosphatidylinositol (GPI) anchor with higher activity. Copyright © 2007 Society of Chemical Industry
DOI: 10.1002/bit.1077
发表时间: 2001-06-20
影响因子: 3.8
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