解淀粉芽孢杆菌
发酵
细菌
生物
微生物
核酸
食品加工中的发酵
放大器
食品科学
生物化学
乳酸
聚合酶链反应
遗传学
基因
作者
Ying Wang,Xuhan Xia,Minghua Wu,Qiyao Sun,Wei Zhang,Yong Qiu,Ruijie Deng,Aimin Luo
标识
DOI:10.1021/acs.jafc.3c04775
摘要
Microorganisms can determine the flavor and quality of fermented food, such as Baijiu, which is produced via Daqu fermentation. Therefore, monitoring key microorganisms during fermentation is important for ensuring high-quality fermented food. Here, we report a single-nucleotide resolved nucleic acid assay based on the CRISPR/Cas12 system, enabling the quantification of Bacillus amyloliquefaciens, a key microorganism in Daqu fermentation at the species level. The assay employs an amplification-refractory mutation system derived from PCR to analyze minor genetic differences between different Bacillus species. The utilization of CRISPR/Cas12 further guaranties the specificity of identifying the PCR amplicon and enables the quantification of Bacillus amyloliquefaciens via end-measurement fluorescence. Compared to conventional qPCR, the assay allows for species-level detection of bacteria, thus enabling the precise detection of the Bacillus strain that yields high-level 2,3,5,6-tetramethylpyrazine. The assay promises the precise monitoring of bacterial growth and contribution to flavor during Daqu fermentation, thus facilitating fermented food quality control.
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