Liu Qi, Song Lili, Peng Qiangqiang, Zhu Qiaoyun, Shi Xiaona, Xu Mingqiang, Wang Qiyao, Zhang Yuanxing, 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, 130 Meilong Road, Shanghai 200237, China.
Sci Adv. 2022 Feb 11;8(6):eabl5166. doi: 10.1126/sciadv.abl5166.
Rapidly growing yeasts with appropriate posttranslational modifications are favored hosts for protein production in the biopharmaceutical industry. However, limited production capacity and intricate transcription regulation restrict their application and adaptability. Here, we describe a programmable high-expression yeast platform, SynPic-X, which responds to defined signals and is broadly applicable. We demonstrated that a synthetic improved transcriptional signal amplification device (iTSAD) with a bacterial-yeast transactivator and bacterial-yeast promoter markedly increased expression capacity in . CRISPR activation and interference devices were designed to strictly regulate iTSAD in response to defined signals. Engineered switches were then constructed to exemplify the response of SynPic-X to exogenous signals. Expression of α-amylase by SynPic-R, a specific SynPic-X, in a bioreactor proved a methanol-free high-production process of recombinant protein. Our SynPic-X platform provides opportunities for protein production in customizable yeast hosts with high expression and regulatory flexibility.
具有适当翻译后修饰的快速生长酵母是生物制药行业生产蛋白质的理想宿主。然而，有限的生产能力和复杂的转录调控限制了它们的应用和适应性。在此，我们描述了一个可编程的高表达酵母平台SynPic-X，它能响应特定信号且具有广泛适用性。我们证明，带有细菌-酵母反式激活因子和细菌-酵母启动子的合成改进转录信号放大装置（iTSAD）显著提高了在……中的表达能力。设计了CRISPR激活和干扰装置，以响应特定信号严格调控iTSAD。然后构建工程开关以例证SynPic-X对外源信号的响应。特定的SynPic-X即SynPic-R在生物反应器中表达α-淀粉酶，证明了重组蛋白的无甲醇高产过程。我们的SynPic-X平台为在具有高表达和调控灵活性的可定制酵母宿主中生产蛋白质提供了机会。
