Department of Biological Systems Engineering, Washington State University, Pullman, Washington 99164, United States.
ACS Synth Biol. 2020 Aug 21;9(8):2208-2213. doi: 10.1021/acssynbio.0c00243. Epub 2020 Jul 9.
Promoters are critical tools to precisely control gene expression for both synthetic biology and metabolic engineering. Although has demonstrated many industrially relevant advantages, promoter discovery efforts on this non-conventional yeast are limited due to the challenge in finding suitable inducible and repressible promoters. Six copper-inducible promoters and five repressible promoters were isolated in this work. Especially, Cu-repressible promoters showed relatively high activity under non-repressing conditions compared with a constitutive promoter, but the strength could be almost fully repressed by a supplement of a low content of Cu. The six Cu-inducible promoters were engineered to improve their dynamic regulation range with a tandem upstream activation sequence. An engineered promoter was successfully used to construct a more productive pathway for production of a novel bioproduct, wax ester, than that used for both Cu-inducible promoter and constitutive promoter. This study provides effective tools applicable to fine-tune the gene expression in this microbial host.
启动子是合成生物学和代谢工程中精确控制基因表达的关键工具。尽管 已经表现出许多工业相关的优势，但由于难以找到合适的诱导和抑制启动子，因此在这种非常规酵母上的启动子发现工作受到限制。本工作中分离了六个铜诱导启动子和五个抑制启动子。特别是，Cu 抑制启动子在非抑制条件下表现出相对较高的活性，与组成型启动子相比，但通过补充低含量的 Cu，强度几乎可以完全抑制。这六个 Cu 诱导启动子被工程改造，通过串联上游激活序列来改善其动态调控范围。一个工程化的启动子被成功地用于构建一个生产新型生物制品——蜡酯的更高效途径，比 Cu 诱导启动子和组成型启动子都更有效。这项研究提供了有效的工具，可用于微调该微生物宿主中的基因表达。
