基于常压室温等离子体诱变的高通量筛选方法提高酿酒酵母中(2S)-柚皮素的产量
Enhancing (2S)-naringenin production in Saccharomyces cerevisiae by high-throughput screening method based on ARTP mutagenesis.
作者信息
Gao Qian, Gao Song, Zeng Weizhu, Li Jianghua, Zhou Jingwen
机构信息
Engineering Research Center of Ministry of Education On Food Synthetic Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122 Jiangsu China.
Science Center for Future Foods, Jiangnan University, 1800 Lihu Rd, Wuxi, 214122 China.
出版信息
3 Biotech. 2024 Mar;14(3):85. doi: 10.1007/s13205-023-03892-6. Epub 2024 Feb 18.
()-Naringenin, a dihydro-flavonoid, serves as a crucial precursor for flavonoid synthesis due to its extensive medicinal values and physiological functions. A pathway for the synthesis of ()-naringenin from glucose has previously been constructed in through metabolic engineering. However, this synthetic pathway of ()-naringenin is lengthy, and the genes involved in the competitive pathway remain unknown, posing challenges in significantly enhancing ()-naringenin production through metabolic modification. To address this issue, a novel high-throughput screening (HTS) method based on color reaction combined with a random mutagenesis method called atmospheric room temperature plasma (ARTP), was established in this study. Through this approach, a mutant (B7-D9) with a higher titer of ()-naringenin was obtained from 9600 mutants. Notably, the titer was enhanced by 52.3% and 19.8% in shake flask and 5 L bioreactor respectively. This study demonstrates the successful establishment of an efficient HTS method that can be applied to screen for high-titer producers of ()-naringenin, thereby greatly improving screening efficiency and providing new insights and solutions for similar product screenings.
()-柚皮素是一种二氢黄酮类化合物,由于其广泛的药用价值和生理功能,它是黄酮类化合物合成的关键前体。此前已通过代谢工程构建了一条从葡萄糖合成()-柚皮素的途径。然而,这条()-柚皮素的合成途径冗长,且参与竞争途径的基因尚不清楚,这给通过代谢修饰显著提高()-柚皮素产量带来了挑战。为了解决这个问题,本研究建立了一种基于颜色反应的新型高通量筛选(HTS)方法,并结合一种名为常压室温等离子体(ARTP)的随机诱变方法。通过这种方法,从9600个突变体中获得了一个()-柚皮素产量更高的突变体(B7-D9)。值得注意的是,在摇瓶和5 L生物反应器中,产量分别提高了52.3%和19.8%。本研究证明成功建立了一种高效的HTS方法,该方法可用于筛选高产()-柚皮素的菌株,从而大大提高筛选效率,并为类似产品的筛选提供新的见解和解决方案。