Promoter-Library-Based Pathway Optimization for Efficient (2S)-Naringenin Production from p-Coumaric Acid in Saccharomyces cerevisiae
Promoter-Library-Based Pathway Optimization for Efficient (2S)-Naringenin Production from p-Coumaric Acid in Saccharomyces cerevisiae
复制标题
基于启动子文库的途径优化，用于在酿酒酵母中从对香豆酸高效生产 (2S)-柚皮素
DOI：
10.1021/acs.jafc.0c01130
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发表时间：
2020-06-24
影响因子：
6.1
通讯作者：
Chen, Jian
中科院分区：
文献类型：
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作者：
Gao, Song;Zhou, Hengrui;Chen, Jian
途径优化在微调代谢途径中起着重要作用。在大多数情况下，超过三个基因参与特定靶产物的生物合成途径。为了提高产品的效价，通过一系列具有不同强度的启动子对一组基因进行合理调控是必不可少的。在一系列RNA-Seq数据的基础上，选择了一组66个天然启动子来微调酿酒酵母中的基因表达。通过荧光激活细胞分选测量增强的绿色荧光蛋白的荧光强度来表征启动子强度。P-TDH 1、P-PGK 1、P-INO 1、P-SED 1和P-CCW 12的表达强于P-TDH 3，而另外15个启动子的表达强于P-TEF 1。在此基础上，选择30个启动子对香豆酸生物合成（2S）-柚皮素的途径进行优化。采用高通量筛选方法，在5L生物反应器中，对香豆酸的（2S）-柚皮素的最高滴度达到1.21 g/L，这是目前报道的最高滴度。
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 Saccharomyces cerevisiae. Promoter strength was characterized by measuring the fluorescence strength of the enhanced green fluorescent protein through fluorescence-activated cell sorting. The expressions of P-TDH1, P-PGK1, P-INO1, P-SED1, and P-CCW12 were stronger than that of P-TDH3, whereas those of another 15 promoters were stronger than that of P-TEF1. Then, 30 promoters were chosen to optimize the biosynthesis pathway of (2S)-naringenin from p-coumaric acid. With a high-throughput screening method, the highest titer of (2S)-naringenin in a 5 L bioreactor reached 1.21 g/L from p-coumaric acid, which is the highest titer according to the currently available reports.