Combinatorial optimization of gene expression through recombinase-mediated promoter and terminator shuffling in yeast

• 全文链接

重组酶介导的酵母启动子和终止子重组基因表达组合优化

关键词：

来源：

Nature Communications

全文链接：

//agri.nais.net.cn/topic/downloadFile/0905f67d-cd61-414c-92c1-bc361d23afdc

来源地址：

https://www.nature.com/articles/s41467-024-44997-7

资源所属：

饲料用酶工程

类型：

学术文献

语种：

英语

原文发布日期：

2024-02-07

摘要：

Microbes are increasingly employed as cell factories to produce biomolecules. This often involves the expression of complex heterologous biosynthesis pathways in host strains. Achieving maximal product yields and avoiding build-up of (toxic) intermediates requires balanced expression of every pathway gene. However, despite progress in metabolic modeling, the optimization of gene expression still heavily relies on trial-and-error. Here, we report an approach for in vivo, multiplexed Gene Expression Modification by LoxPsym-Cre Recombination (GEMbLeR). GEMbLeR exploits orthogonal LoxPsym sites to independently shuffle promoter and terminator modules at distinct genomic loci. This approach facilitates creation of large strain libraries, in which expression of every pathway gene ranges over 120-fold and each strain harbors a unique expression profile. When applied to the biosynthetic pathway of astaxanthin, an industrially relevant antioxidant, a single round of GEMbLeR improved pathway flux and doubled production titers. Together, this shows that GEMbLeR allows rapid and efficient gene expression optimization in heterologous biosynthetic pathways, offering possibilities for enhancing the performance of microbial cell factories.