Engineering Saccharomyces boulardii for Probiotic Supplementation of l‐Ergothioneine

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 stud...

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Bibliographic Details
Published in:Biotechnology journal Vol. 19; no. 11; pp. e202400527 - n/a
Main Authors: Tang, Chaoqun, Zhang, Lu, Wang, Junyi, Zou, Congjia, Zhang, Yalin, Yuan, Jifeng
Format: Journal Article
Language:English
Published: Germany 01.11.2024
Subjects:
antioxidant
Antioxidants
Ergothioneine
Gene Editing - methods
Genetic Engineering - methods
l‐ergothioneine
PiggyBac transposon
Probiotics
Saccharomyces boulardii
Saccharomyces boulardii - genetics
Saccharomyces boulardii
antioxidant
l‐ergothioneine
PiggyBac transposon
ISSN:1860-6768, 1860-7314, 1860-7314
Online Access:Get full text
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Summary: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 a 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. Graphical and Lay Summary Saccharomyces boulardii, as a probiotic yeast, has shown great potential in regulating gut health and treating gastrointestinal diseases. In this study, 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 PiggyBac transposon system, a great variety of EGT‐producing strains was obtained, and one best isolated top producer reached 17.50 mg/L EGT after 120 h cultivation. In the future, the engineered S. boulardii is expected to have potential applications for probiotic supplementation of antioxidants to improve the human health.
Bibliography:Funding
This study was funded by National Natural Science Foundation of China 32071030, 32270087, Xiamen University 20720240120.
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ISSN:1860-6768
1860-7314
1860-7314
DOI:10.1002/biot.202400527