Pyrococcus furiosus argonaute based Alicyclobacillus acidoterrestrsis detection in fruit juice

Yiheng Shi, Zishan Tan, Di Wu, Yongning Wu, Guoliang Li*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Abstract

Alicyclobacillus acidoterrestris is the major threat to fruit juice for its off-odor producing characteristic. In this study, Pyrococcus furiosus Argonaute (PfAgo), a novel endonuclease with precise DNA cleavage activity, was used for A. acidoterrestris detection, termed as PAD. The partially amplified 16S rRNA gene of A. acidoterrestris can be cleaved by PfAgo activated by a short 5′-phosphorylated single strand DNA, producing a new guide DNA (gDNA). Then, PfAgo was activated by the new gDNA to cut a molecular beacon (MB) with fluorophore-quencher reporter, resulting in the recovery of fluorescence. The fluorescent intensity is positively related with the concentration of A. acidoterrestris. The PAD assay showed excellent specificity and sensitivity as low as 101 CFU/mL, which can be a powerful tool for on-site detection of A. acidoterrestris in fruit juice industry in the future, reducing the economic loss.
Original languageEnglish
Article number104475
Number of pages6
JournalFood Microbiology
Volume120
Early online date15 Jan 2024
DOIs
Publication statusPublished - 01 Jun 2024

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