Development of Siderophore-Based Fluorescent Probe for Vibrios Detection

• Main Authors: Peng-Hsun Chen, 陳芃勳
• Other Authors: 王宗興
• Format: Others
• Language: en_US
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/45574211816789196431

碩士 === 國立臺灣大學 === 化學研究所 === 104 === Vibrio is a genus of gram-negative bacteria that is highly relevant to our life. Given that illnesses caused by these microbes have escalating globally, it behooves us to develop a Vibrio-specific probe. To this end, we take our aim at iron acquisition mechanism mediated by siderophore, a small molecule used to specifically transport iron into bacteria under iron-limited condition. By hijacking the pathway, we may find us an efficient and specific way to label Vibrio spp. Herein we first functionalize vibrioferrin (VF), a siderophore secreted by V. parahaemolyticus. Then an alkyne-containing linker is harbored onto this siderophore analog to undergo copper catalyzed azide-alkyne cycloaddition with an azdio-containing fluorescein. On flow cytometry, upon treatment with our probe, VSP1, under iron-limited conditions, significant fluorescent increases can be found not only in its secretor, V. parahaemolyticus but also in other vibrio spp., V. cholerae and V. vulnificus, while no differences are observed in S. aureus, B. subtilis, E. coli and S. enterica. On the contract, when in an iron-rich media, no enhancements are observed in the bacteria tested. The strain dependency is also demonstrated on fluorescent images, where vibrios can be labelled, while S. aureus and E. coli give no response. The results suggest that our probe has the potential for specific labelling vibrios under ironlimited condition. In addition, the unexpected outcome given by V. cholerae and V. vulnificus, indicating VF may be an undiscovered xenosiderophore.