Liu Qi, He Ziyu, Cai Menghao
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.
Shanghai Collaborative Innovation Center for Biomanufacturing, Shanghai, 200237, China.
Synth Syst Biotechnol. 2024 Sep 23;10(1):133-139. doi: 10.1016/j.synbio.2024.09.008. eCollection 2025.
Methanol is a promising substrate for sustainable biomanufacturing, and has become a commonly used yeast for methanol utilization due to its powerful methanol metabolic pathways and methanol inducible promoter. Previous reconstruction of gene circuits highly improved transcriptional activity, but excessive expression of chimeric transactivator damaged cell growth on methanol. Here we employed transcriptome analysis to investigate the effects of chimeric transactivator overexpression on cellular metabolism and regulatory networks. The results showed that strong expression of chimeric transactivator unexpectedly downregulated methanol metabolism, especially the (), but without remarkable changes in expression of transcriptional factors. Meanwhile, the synthesis of peroxisomes also varied with chimeric transactivator expression. In addition, the enrichment analysis of differentially expressed genes revealed their impact on cellular metabolism. The gene expression patterns caused by different expression levels of chimeric transactivators have also been clarified. This work provides useful information to understand the transcriptional regulation of the promoter and methanol signaling. It revealed the importance of balancing transcription factor expression for the host improvement.
甲醇是可持续生物制造中一种很有前景的底物，由于其强大的甲醇代谢途径和甲醇诱导型启动子，已成为一种常用的甲醇利用酵母。先前基因回路的重建极大地提高了转录活性，但嵌合反式激活因子的过度表达损害了细胞在甲醇上的生长。在这里，我们采用转录组分析来研究嵌合反式激活因子过表达对细胞代谢和调控网络的影响。结果表明，嵌合反式激活因子的强表达意外地下调了甲醇代谢，尤其是（此处原文缺失具体内容），但转录因子的表达没有显著变化。同时，过氧化物酶体的合成也随嵌合反式激活因子的表达而变化。此外，差异表达基因的富集分析揭示了它们对细胞代谢的影响。由不同表达水平的嵌合反式激活因子引起的基因表达模式也已得到阐明。这项工作为理解启动子的转录调控和甲醇信号传导提供了有用信息。它揭示了平衡转录因子表达对宿主改良的重要性。
