Development of a nitrate-responsive whole-cell biosensor based on engineered probiotic Saccharomyces boulardii

IF 2.4 3区农林科学 Q3 FOOD SCIENCE & TECHNOLOGY

Food Science and Biotechnology Pub Date : 2025-03-12 DOI:10.1007/s10068-025-01850-x

Woohyuk Lee, Sang-Hun Do, Jonghyeok Shin, Sun-Ki Kim

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Abstract

This study aimed to construct a probiotic Saccharomyces boulardii-based biosensor that can sense and respond to nitrate, a biomarker of gut inflammation. To this end, a portion of the nitrate assimilation pathway in Ogataea polymorpha required for the designed nitrate-responsive genetic circuit was introduced into S. boulardii, which lacks the corresponding pathway. Specifically, the genes involved in nitrate transport (YNT1) and nitrate-responsive transcriptional activation (YNA1 and YNA2) were introduced into S. boulardii using the CRISPR/Cas9-based genome editing system to construct the SKSC499 strain. Next, we investigated whether YNT1, YNI1, and YNR1 promoters, which are activated by nitrate-bound YNA1/YNA2 transcription factor in O. polymorpha, induced expression of the reporter gene (G418 resistance marker) in the SKSC499 strain in a nitrate concentration-dependent manner. Among the candidate promoters, the YNR1 promoter effectively induced growth of the SKSC499 strain only when nitrate was present in the medium containing the G418 antibiotic.

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基于工程益生菌博氏酵母菌的硝酸盐响应全细胞生物传感器的研制

本研究旨在构建一种基于博拉迪酵母菌的益生菌生物传感器，该传感器可以感知并响应肠道炎症的生物标志物硝酸盐。为此，将设计的硝酸盐响应遗传回路所需的多态Ogataea （Ogataea polymorpha）硝酸盐同化途径的一部分引入到S. boulardii中，后者缺乏相应的途径。具体而言，利用基于CRISPR/ cas9的基因组编辑系统，将参与硝酸盐转运（YNT1）和硝酸盐应答转录激活（YNA1和YNA2）的基因导入博氏沙门氏菌，构建SKSC499菌株。接下来，我们研究了被多态稻中硝酸盐结合的YNA1/YNA2转录因子激活的YNT1、YNI1和YNR1启动子是否以硝酸盐浓度依赖的方式诱导SKSC499菌株中报告基因（G418抗性标记）的表达。在候选启动子中，只有在含有G418抗生素的培养基中存在硝酸盐时，YNR1启动子才能有效诱导SKSC499菌株的生长。

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来源期刊

Food Science and Biotechnology FOOD SCIENCE & TECHNOLOGY-

CiteScore

5.40

自引率

3.40%

发文量

174

审稿时长

2.3 months

期刊介绍： The FSB journal covers food chemistry and analysis for compositional and physiological activity changes, food hygiene and toxicology, food microbiology and biotechnology, and food engineering involved in during and after food processing through physical, chemical, and biological ways. Consumer perception and sensory evaluation on processed foods are accepted only when they are relevant to the laboratory research work. As a general rule, manuscripts dealing with analysis and efficacy of extracts from natural resources prior to the processing or without any related food processing may not be considered within the scope of the journal. The FSB journal does not deal with only local interest and a lack of significant scientific merit. The main scope of our journal is seeking for human health and wellness through constructive works and new findings in food science and biotechnology field.