通过去除n -糖基化改善重组毕赤酵母抗冻蛋白的热滞活性

Eun Jae Kim, J. Lee, S. G. Lee, S. Han
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引用次数: 6

摘要

为了在零下环境中生存,极地生物产生冰结合蛋白(IBPs)。这些ibp可以防止细胞内形成大的冰晶,这对生物体可能是致命的。最近,在毕氏酵母中克隆并生产了一株重组FfIBP(来自frigoris Flavobacterium PS1)。在这项研究中,我们证明了P. pastoris产生的FfIBP具有糖基化位点,这降低了FfIBP的热滞后活性。pastoris表达的FfIBP在SDS-PAGE上呈现双链。糖苷酶反应结果表明,FfIBP具有复杂的n -连接低聚糖。这些结果表明糖基化位点的残基可能会干扰FfIBP与冰分子的结合。本研究结果可用于大规模生产高活性的抗冻蛋白。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Improving thermal hysteresis activity of antifreeze protein from recombinant Pichia pastoris by removal of N-glycosylation
ABSTRACT To survive in a subzero environment, polar organisms produce ice-binding proteins (IBPs). These IBPs prevent the formation of large intracellular ice crystals, which may be fatal to the organism. Recently, a recombinant FfIBP (an IBP from Flavobacterium frigoris PS1) was cloned and produced in Pichia pastoris using fed-batch fermentation with methanol feeding. In this study, we demonstrate that FfIBP produced by P. pastoris has a glycosylation site, which diminishes the thermal hysteresis activity of FfIBP. The FfIBP expressed by P. pastoris exhibited a doublet on SDS-PAGE. The results of a glycosidase reaction suggested that FfIBP possesses complex N-linked oligosaccharides. These results indicate that the residues of the glycosylated site could disturb the binding of FfIBP to ice molecules. The findings of this study could be utilized to produce highly active antifreeze proteins on a large scale.
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