National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
J Agric Food Chem. 2020 Jun 24;68(25):6884-6891. doi: 10.1021/acs.jafc.0c01130. Epub 2020 Jun 11.
Pathway optimization plays an important role in fine-tuning metabolic pathways. In most conditions, more than three genes are involved in the biosynthesis pathway of a specific target product. To improve the titer of products, rational regulation of a group of genes by a series of promoters with different strengths is essential. On the basis of a series of RNA-Seq data, a set of 66 native promoters was chosen to fine-tune gene expression in . Promoter strength was characterized by measuring the fluorescence strength of the enhanced green fluorescent protein through fluorescence-activated cell sorting. The expressions of P, P, P, P, and P were stronger than that of P, whereas those of another 15 promoters were stronger than that of P. Then, 30 promoters were chosen to optimize the biosynthesis pathway of (2)-naringenin from -coumaric acid. With a high-throughput screening method, the highest titer of (2)-naringenin in a 5 L bioreactor reached 1.21 g/L from -coumaric acid, which is the highest titer according to the currently available reports.
通路优化在精细调控代谢通路方面起着重要作用。在大多数情况下，特定目标产物的生物合成途径涉及三个以上的基因。为了提高产物的产量，需要通过一系列强度不同的启动子对一组基因进行合理的调控。基于一系列 RNA-Seq 数据，选择了 66 个天然启动子来微调. 通过荧光激活细胞分选测量增强型绿色荧光蛋白的荧光强度来表征启动子强度。P、P、P、P 和 P 的表达强于 P，而另外 15 个启动子的表达强于 P。然后，选择 30 个启动子来优化从 -香豆酸到（2）-柚皮素的生物合成途径。采用高通量筛选方法，在 5 L 生物反应器中从 -香豆酸获得的（2）-柚皮素的最高产量达到 1.21 g/L，这是根据现有报道获得的最高产量。
