The methylotrophic yeast Pichia pastoris is well-known for the production of a broad spectrum of functional types of heterologous proteins including enzymes, antigens, engineered antibody fragments, and next gen protein scaffolds and many transcription factors are utilized to address the burden caused by the high expression of heterologous proteins. In this article, a novel P. pastoris transcription factor currently annotated as Fhl1p, an activator of ribosome biosynthesis processing, was investigated for promoting the expression of the recombinant proteins. The function of Fhl1p of P. pastoris for improving the expression of recombinant proteins was verified in strains expressing phytase, pectinase and mRFP, showing that the productivity was increased by 20–35%. RNA-Seq was used to study the Fhl1p regulation mechanism in detail, confirming Fhl1p involved in the regulation of rRNA processing genes, ribosomal small/large subunit biogenesis genes, Golgi vesicle transport genes, etc., which contributed to boosting the expression of foreign proteins. The overexpressed Fhl1p strain exhibited increases in the polysome and monosome levels, showing improved translation activities. This study illustrated that the transcription factor Fhl1p could effectively enhance recombinant protein expression in P. pastoris. Furthermore, we provided the evidence that overexpressed Fhl1p was related to more active translation state.

中文翻译：

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巴斯德毕赤酵母中的正转录因子Fhl1p蛋白增强重组蛋白的表达

甲基营养酵母巴斯德毕赤酵母因产生多种功能类型的异源蛋白质而广为人知，这些异源蛋白质包括酶，抗原，工程抗体片段和下一代蛋白质支架，并利用许多转录因子来解决由高蛋白引起的负担异源蛋白的表达。在本文中，研究了一种新的巴斯德毕赤酵母转录因子，目前被称为Fhl1p（核糖体生物合成过程的激活剂），以促进重组蛋白的表达。在表达植酸酶，果胶酶和mRFP的菌株中证实了巴斯德毕赤酵母Fhl1p改善重组蛋白表达的功能，表明生产力提高了20-35％。RNA-Seq用于详细研究Fhl1p调控机制，证实Fhl1p参与了rRNA加工基因，核糖体小/大亚基生物发生基因，高尔基囊泡转运基因等的调控，从而促进了外源蛋白的表达。过表达的Fhl1p菌株在多核糖体和单核糖体水平上表现出增加，显示出改善的翻译活性。这项研究表明，转录因子Fhl1p可以有效地增强巴斯德毕赤酵母中的重组蛋白表达。此外，我们提供了证据，表明Fhl1p的过表达与更活跃的翻译状态有关。过表达的Fhl1p菌株在多核糖体和单核糖体水平上表现出增加，显示出改善的翻译活性。这项研究表明，转录因子Fhl1p可以有效地增强巴斯德毕赤酵母中的重组蛋白表达。此外，我们提供了证据，表明Fhl1p的过表达与更活跃的翻译状态有关。过表达的Fhl1p菌株在多核糖体和单核糖体水平上表现出增加，显示出改善的翻译活性。这项研究表明，转录因子Fhl1p可以有效地增强巴斯德毕赤酵母中的重组蛋白表达。此外，我们提供了证据，表明Fhl1p的过表达与更活跃的翻译状态有关。