Enhancing the Secretion Efficiency of Heterologous Proteins in Pichia pastoris through N-Glycosylation and SOD1-Mediated Redox Regulation

This project presents the RNA seq data generated from two engineered strains of the methylotrophic yeast Pichia pastoris GS115, which exhibit differential secretion efficiencies for enhanced green fluorescent protein. These strains, designated GS115-High and GS115-Low, were constructed through site-directed mutagenesis of glycosylation sites within the alpha-mating factor signal peptide. The primary objective of this study was to investigate the global transcriptional mechanisms underlying divergent protein secretion capacities. Comparative transcriptomic analysis between the high and low secretion strains revealed significant upregulation of the superoxide dismutase gene and enhanced oxidoreductase activity in the GS115-High strain, suggesting a critical link between efficient protein secretion and robust cellular antioxidant defense. Additionally, pathways related to amino acid metabolism were notably altered, indicating a synergistic reallocation of metabolic resources. This dataset provides valuable insights into the cooperative interplay between protein processing and redox homeostasis, serving as a key resource for understanding and engineering P. pastoris for improved heterologous protein production.

本项目呈现了两株工程化甲醇营养型酵母毕赤酵母（Pichia pastoris）GS115的RNA测序数据，二者对增强型绿色荧光蛋白（enhanced green fluorescent protein）的分泌效率存在显著差异。这两株菌株分别命名为GS115-High与GS115-Low，通过对α-交配因子信号肽（alpha-mating factor signal peptide）内的糖基化位点进行定点突变构建获得。本研究的核心目标为探究驱动蛋白质分泌能力分化的全局转录调控机制。通过对高、低分泌效率菌株开展比较转录组学分析，研究人员发现GS115-High菌株中超氧化物歧化酶（superoxide dismutase）基因显著上调，且氧化还原酶（oxidoreductase）活性增强，提示高效蛋白质分泌与强健的细胞抗氧化防御系统之间存在关键关联。此外，与氨基酸代谢（amino acid metabolism）相关的通路亦发生显著改变，表明代谢资源发生了协同性重分配。本数据集为解析蛋白质加工与氧化还原稳态（redox homeostasis）之间的协同互作提供了宝贵见解，同时可作为理解并工程化改造毕赤酵母以提升异源蛋白生产效率的关键资源。