期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 68, 期 25, 页码 6884-6891出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.0c01130
关键词
promoter; fine-tuning; metabolic pathway; (2S)-naringenin; yeast
资金
- National Key Research and Development Program of China [2017YFC1600403]
- National Outstanding Youth Foundation [21822806]
- National Natural Science Foundation of China [31670095, 31770097]
- Distinguished Professor Project of Jiangsu Province
- Fundamental Research Funds for the Central Universities [JUSRP51701A]
- 111 Project [111-2-06]
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.
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