| Summary: | 碩士 === 國立臺北科技大學 === 生物科技研究所 === 98 === Nitroreductase (NTR) is a flavoenzyme that catalyzes the reduction of aromatic nitro compound to correspond in aromatic amine. The NTR is widely utilized for the activating the prodrugs in cancer therapy and in the biosensor applications for detecting the environmental nitroaromatics. We prepared a new long-wavelength latent fluorogenic probe, NRLF2, for the real time monitoring NRT activities. Latent fluorophores are stable probes that unmask their intense fluorescence only by a user-designated chemical reaction, and they are especially useful tools for basic research in the biological sciences. The fluorogenic chemical transformation of NRLF2 triggered by NTR in the presence of NADH is through a series of tandem reactions which are spontaneous and irreversible at physiological temperature in aqueous media. The fluorescent response of NRLF2 is insensitive to various biological and chemical reductants. The fluorescence signal revealed by this is specific and exhibited the near infrared spectrum region with emission maximum at 595 nm, and it could be competitively inhibited by aromatic nitro group. NRLF2 coupled with NRT is a “switch off” flourescence probe for the rapid screening nitro aromatic compounds. This novel fluorimetric probe demonstrates to be a rapid sensor in detecting the nitroaromatics of environmental pollutants. This the oxygen-insensitive assay demonstrates a good relationship in detecting 3-hydroxybutyrate and propionaldehyde in the 4–20 μM and 0-150 μM range respectively, which present to the applicability as a biosensor for future clinical diagnosis and inspecting pollutants.
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