次生菌是头孢菌素C(CPC)的主要工业生产者,用作生产重要的头孢菌素类抗生素的原料。由于缺乏多样化的启动子元件,代谢工程转化的发展相对缓慢,导致CPC生产的改进有限。在这项研究中,基于对转录组概况的分析,选择27个候选启动子来驱动报道基因的表达。该文库的启动子活性为对照启动子PAngpdA的0.0075至101倍。同时,开发了一种快速筛选潜在双向启动子的方法,并鉴定并验证了来自27个候选选项的4个强双向启动子.最后,使用金门方法将来自文库的启动子模块与各种靶基因组合。通过混合转化和筛选过程,鉴定了高产菌株AG-6、AG-18和AG-41,CPC产量增长30%,35%,29%,分别,与对照菌株Ac-Δaxl2::eGFP相比。因此,该启动子库的利用提供了更广泛的合成生物学工具包,用于黄A.从而为基因表达的精确调控奠定了坚实的基础。
Acremonium chrysogenum is the major industrial producer of cephalosporin C (CPC), which is used as raw material for the production of significant cephalosporin antibiotics. Due to the lack of diverse promoter elements, the development of metabolic engineering transformation is relatively slow, resulting in a limited improvement on CPC production. In this study, based on the analysis of the transcriptome profile, 27 candidate promoters were selected to drive the expression of the reporter genes. The promoter activities of this library ranged from 0.0075 to 101 times of the control promoter PAngpdA . Simultaneously, a rapid screening method for potential bidirectional promoters was developed and 4 strong bidirectional promoters from 27 candidate options were identified and validated. Finally, the Golden Gate method was employed to combine promoter modules from the library with various target genes. Through a mixed transformation and screening process, high-yielding strains AG-6, AG-18, and AG-41 were identified, exhibiting an increase in CPC production of 30%, 35%, and 29%, respectively, compared to the control strain Ac-∆axl2:: eGFP. Therefore, the utilization of this promoter library offers a broader range of synthetic biology toolkits for the genetic engineering transformation of A. chrysogenum, thus establishing a solid foundation for the precise regulation of gene expression.