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首页> 外文期刊>Journal of Biotechnology >KlPMR1 inactivation and calcium addition enhance secretion of non-hyperglycosylated heterologous proteins in Kluyveromyces lactis
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KlPMR1 inactivation and calcium addition enhance secretion of non-hyperglycosylated heterologous proteins in Kluyveromyces lactis

机译：KlPMR1失活和钙添加增强乳酸克鲁维酵母中非高糖基化异源蛋白的分泌

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摘要

The Kluyveromyces lactis K1PMR1 gene is the functional homologue of Saccharomyces cerevisiae PMR1 which encodes a Ca2+-ATPase localized in the Golgi apparatus. We studied the effects of K1PMR1 inactivation on the glycosylation and secretion of native and heterologous proteins in K. lactis. We used acid phosphatase, recombinant human serum albumin and alpha-glucoamylase from Arxula adeninivorans as reporter proteins. The Klpmrl Delta strain showed enhanced secretion of the heterologous proteins analyzed; the improved rHSA production did not result from enhanced transcription but rather involved increased translation and/or secretion efficiency. The growth rate of mutant cells resulted slower as compared to that of wild-type strain. The addition of 10 mM calcium to the culture medium, however, not only completely relieved the growth defect of the mutant cells but also improved the rate of heterologous proteins production. Moreover, the addition of this ion in the culture medium of K. lactis did not suppress the glycosylation defects; this is an important difference with respect to S. cerevisiae where the glycosylation is partially restored by Ca2+ addition. The Klpmrl Delta strain as a host offers thus an additional advantage for those cases requiring that the produced recombinant protein would not result hyperglycosylated
机译：乳酸克鲁维酵母K1PMR1基因是酿酒酵母PMR1的功能同源物，其编码位于高尔基体中的Ca2 + -ATPase。我们研究了K1PMR1失活对乳酸克鲁维酵母中糖基化和天然和异源蛋白分泌的影响。我们将酸性磷酸酶，重组人血清白蛋白和来自Arxula adeninivorans的α-葡糖淀粉酶用作报告蛋白。 Klpmrl Delta菌株显示出所分析异源蛋白质的分泌增强; rHSA产量的提高不是由于转录增强，而是翻译和/或分泌效率提高。与野生型菌株相比，突变细胞的生长速率较慢。但是，向培养基中添加10 mM钙不仅可以完全缓解突变细胞的生长缺陷，而且可以提高异源蛋白质的产生速度。此外，在乳酸克鲁维酵母的培养基中加入该离子不能抑制糖基化缺陷。对于酿酒酵母而言，这是重要的区别，在酿酒酵母中，通过添加Ca2 +可部分恢复糖基化。因此，Klpmrl Delta菌株作为宿主为那些需要产生的重组蛋白不会导致高糖基化的情况提供了额外的优势

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