PDF(Pubmed)
Abstract：
Komagataella phaffii (previously described as Pichia pastoris) is a yeast that produces high-level heterologous proteins with a wide range of applications in medicine and industry. The methanol-induced alcohol oxidase I promoter (PAOX1) is frequently used for protein expression in this yeast. However, limitations on the use of methanol have been observed in large-scale production, including its flammability, toxicity, and need for special handling. Here, we propose to develop a system using recombinant cells constitutively expressing pectinmethyl esterase for expression of two reporter proteins, GFP and azurin, under the control of PAOX1 using pectin in production medium. So, this system is coherent with yeast culture medium containing pectin and heterologous gene inserted downstream of PAOX1 can be successfully expressed without the addition of methanol. Therefore, this novel Self-inducibLe heterologous protein EXpression (SILEX) system, which does not require the addition of methanol, can be used for the production of any protein. It can also be adapted for large-scale production.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s13205-024-04039-x.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s13205-024-04039-x.
摘要：
Komagataellaphaffii（先前描述为巴斯德毕赤酵母）是一种酵母，可产生高水平的异源蛋白质，在医学和工业中具有广泛的应用。甲醇诱导的醇氧化酶I启动子（PAOX1）通常用于该酵母中的蛋白质表达。然而,在大规模生产中观察到使用甲醇的限制，包括它的可燃性,毒性,需要特殊处理。这里,我们建议开发使用重组细胞组成型表达果胶甲基酯酶的系统，用于表达两种报告蛋白，GFP和天青素，在PAOX1的控制下，在生产培养基中使用果胶。所以,该系统与含有果胶的酵母培养基相干，在PAOX1下游插入异源基因，无需添加甲醇即可成功表达。因此,这种新型的自诱导异源蛋白表达（SILEX）系统，不需要添加甲醇，可用于生产任何蛋白质。它也可以适用于大规模生产。
■在线版本包含补充材料，可在10.1007/s13205-024-04039-x获得。
■在线版本包含补充材料，可在10.1007/s13205-024-04039-x获得。
