Engineering Saccharomyces boulardii for probiotic supplementation of L-ergothioneine
Chaoqun Tang1,2, Lu Zhang1, Junyi Wang1, Congjia Zou1, Yalin Zhang1* and Jifeng Yuan1,3*
1 State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University, Fujian 361102, China
2 Department of Basic Medical Sciences, Qinghai University Medical College, Xining, Qinghai 810016, China
3 Shenzhen Research Institute of Xiamen University, Shenzhen 518057, China
* Corresponding author address: State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University, Fujian, China 361102. Email address: zhangyalin@xmu.edu.cn or jfyuan@xmu.edu.cn
Abstract
Saccharomyces boulardii, as a probiotic yeast, has shown great potential in regulating gut health and treating gastrointestinal diseases. Due to its unique antimicrobial and immune-regulating functions, it has become a significant subject of research in the field of probiotics. In this study, we aim to enhance the antioxidant properties of S. boulardii by producing L-ergothioneine (EGT). We first constructed double knockout of ura3 and trp1 gene in S. boulardii, to facilitate plasmid-based expressions. To further enable effective genome editing of S. boulardii, we implemented the PiggyBac system to transpose the heterologous gene expression cassettes into the chromosomes of S. boulardii. By using enhanced green fluorescent protein (EGFP) as the reporter gene, we achieved random chromosomal integration of EGFP expression cassette. By using PiggyBac transposon system, a great variety of EGT-producing strains was obtained, which is not possible for the conventional single target genome editing, and one best isolated top producer reached 17.50 mg/L EGT after 120 h cultivation. In summary, we have applied the PiggyBac transposon system to S. boulardii for the first time for genetic engineering. The engineered probiotic yeast S. boulardii has been endowed with new antioxidant properties and produces EGT. It has potential applications in developing novel therapeutics and dietary supplements for the prevention and treatment of gastrointestinal disorders.
Key words: Saccharomyces boulardii; PiggyBac transposon; antioxidant; L-ergothioneine
全文连接: doi:10.1002/biot.202400527