Boosting 11-oxo-ß-amyrin and glycyrrhetinic acid synthesis in Saccharomyces cerevisiae via pairing novel oxidation and reduction system from legume plants

Glycyrrhetinic acid (GA) and its precursor, 11-oxo-ss-amyrin, are typical triterpenoids found in the roots of licorice, a traditional Chinese medicinal herb that exhibits diverse functions and physiological effects. In this study, we developed a novel and highly efficient pathway for the synthesis of GA and 11-oxo-ss-amyrin in Saccharomyces cerevisiae by introducing efficient cytochrome P450s (CYP450s: Uni25647 and CYP72A63) and pairing their reduction systems from legume plants through transcriptome and genome-wide screening and identification. By increasing the copy number of Uni25647 and pairing cytochrome P450 reductases (CPRs) from various plant sources, the titers of 11-oxo-ss-amyrin and GA were increased to 108.1 +/- 4.6 mg/L and 18.9 +/- 2.0 mg/L, which were nearly 1422-fold and 946.5-fold higher, respectively, compared with previously reported data. To the best of our knowledge, these are the highest titers reported for GA and 11-oxo-ss-amyrin from S. cerevisiae, indicating an encouraging and promising approach for obtaining increased GA and its related triterpenoids without destroying the licorice plant or the soil ecosystem.

甘草次酸（GA）及其前体11 - 氧 - β - 香树脂醇是在甘草根中发现的典型三萜类化合物，甘草是一种具有多种功能和生理效应的传统中药材。在本研究中，我们通过转录组和全基因组筛选与鉴定，引入高效的细胞色素P450（CYP450：Uni25647和CYP72A63）并搭配来自豆科植物的还原系统，在酿酒酵母中开发了一种合成GA和11 - 氧 - β - 香树脂醇的新型高效途径。通过增加Uni25647的拷贝数并搭配来自不同植物来源的细胞色素P450还原酶（CPR），11 - 氧 - β - 香树脂醇和GA的滴度分别提高到108.1 ± 4.6毫克/升和18.9 ± 2.0毫克/升，与先前报道的数据相比，分别提高了近1422倍和946.5倍。据我们所知，这是酿酒酵母中GA和11 - 氧 - β - 香树脂醇所报道的最高滴度，这表明了一种在不破坏甘草植物或土壤生态系统的情况下获取更多GA及其相关三萜类化合物的令人鼓舞且有前景的方法。