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.

Chaoqun Tang

Lu Zhang

Junyi Wang

Congjia Zou

Yalin Zhang

Jifeng Yuan

Biotechnology Journal

BIOTECHNOLOGY JOURNAL (2019 JOURNAL CITATION REPORTS: 3.543) 范围全面，发表经过严格同行评审的论文，涵盖生物技术所有领域的新颖方面和方法。一些问题专门针对一个特殊主题，提供有关研究和技术进步的最关键领域的最新信息。除了这些特殊问题之外，该杂志还欢迎未经请求提交的主要研究文章，例如研究文章、快速通信和生物技术方法。 BTJ也欢迎REVIEW ARTICLES的提案-请将文章的简要概述和资深作者的简历发送给编辑部。BTJ特别强调：系统生物技术合成生物学和代谢工程纳米生物技术和生物材料组织工程，再生医学和干细胞细胞基因编辑、基因治疗和免疫治疗组学技术工业生物技术、生物制药和生物催化生物过程工程和下游加工植物生物技术生物安全、生物技术伦理、科学传播方法和进展。

The Biotechnology Journal is a comprehensive journal covering all areas of biotechnology, publishing high-quality research papers that undergo rigorous peer review. The journal welcomes special issues focused on specific topics, as well as unsolicited submissions of primary research articles, including research papers, rapid communications, and biotechnology methods. We particularly encourage proposals for review articles, requiring authors to submit a brief overview of the article along with the CV of the senior author to the editorial office. The journal emphasizes areas such as systems biotechnology, synthetic biology, metabolic engineering, nanobiotechnology, biomaterials, tissue engineering, regenerative medicine, stem cells, cell gene editing, gene therapy, immunotherapy, omics technologies, industrial biotechnology, biopharmaceuticals, biocatalysis, bioprocess engineering, downstream processing, plant biotechnology, biosafety, biotechnology ethics, and scientific communication.

《Biotechnology Journal》是一份涵盖生物技术各个领域的综合性期刊，经过严格的同行评审，发表高质量的研究论文。该期刊不仅欢迎关于特定主题的专刊，还接受未经请求的主要研究文章，包括研究论文、快速通讯和生物技术方法。我们特别鼓励提交综述文章的提案，作者需向编辑部提供文章的简要概述及资深作者的简历。期刊特别关注的领域包括：系统生物技术、合成生物学、代谢工程、纳米生物技术、生物材料、组织工程、再生医学、干细胞、细胞基因编辑、基因治疗、免疫治疗、组学技术、工业生物技术、生物制药、生物催化、生物过程工程、下游加工、植物生物技术、生物安全、生物技术伦理及科学传播等。

Category	Quartile
生物学	3
生物学, 生化研究方法	3
生物学, 生物工程与应用微生物	3

Engineering Saccharomyces boulardii for Probiotic Supplementation of l‐Ergothioneine

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