Fluorescent probe for detecting nitroreductase and preparation method and application thereof in enzymatic reaction

Abstract

The invention discloses a fluorescent probe for detecting NTR (nitroreductase) and a preparation method and application thereof in enzymatic reaction, and belongs to the technical field of industrialanalysis detection. The fluorescent probe is a 3-(4-(2-(4'-(diphenylamino)-3-((4-nitrobenzyl)oxy)-[1,1'-biphenyl]-4-yl)vinyl)quinoline-1-bromo)propane-1-sulfonate. The fluorescent probe compound has the advantages that by leading hydrophilic group sulfonate and quinoline salt, the hydrophilia is enhanced; under the catalyzing function of NTR, the 1,6-rearrangement and elimination reaction is performed, the hydroxyl is produced, and the detection and analysis of NTR in the enzymatic reaction are realized by the fluorescence change caused by ICT (intramolecular charge transfer) effect. The preparation method has the advantages including that the preparation is simple and convenient, and the yield is high; the preparation method is suitable for detection of content of high-concentration enzyme in the enzymatic reaction, and the great application prospect is realized in the field of detection of enzyme in the enzymatic reaction system in the chemical engineering field.

Description

technical field [0001] The invention belongs to the technical field of industrial analysis and detection, and in particular relates to a fluorescent probe for detecting nitroreductase, a preparation method thereof and an application of enzymatic reaction. Background technique [0002] Nitro compounds are widely used in industries such as medicine, dyes, pesticides and explosives, but most of them are carcinogenic to humans, causing many diseases and endangering human health. Amine compounds are very important in the synthesis of many fine chemical products and intermediates such as pesticides, medicines, dyes, synthetic resins, surfactants, etc., because the introduction of amino groups makes it possible to change the functions of fine chemicals, such as amino groups. The introduction can red-shift the absorption and emission spectra of the compound, and the introduction of amino groups in the ortho position of the dye chromophore can cause the color change of the dye, and t...

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Problems solved by technology

Chinese patent CN201610050741.X prepared a two-photon fluorescent probe for detecting nitroreductase in the anoxic region. The aromatic nitro group in the compound can be reduced to an aromatic amine group by the nitroreductase and trigger 1,6 - Rearrangement and elimination reactions release the fluorescent host, resulting in a change in fluorescence, but the fluorescent probe is generally water-soluble and has the property of aggregation-induced quenching, which makes it difficult to achieve high concentration and enzyme detection analysis in the aqueous phase. Simultaneous use Two-photon detection equipment is complex and expensive, and its application areas are mainly concentrated in hypoxic regions in cells

Chinese patent CN201610471060.0 discloses a two-photon fluorescent probe that can detect nitroreductase. The nitro group is directly connected to the fluorescent body. As the concentration of nitroreductase increases, the fluorescence intensity gradually increases, and the emission wavelength range Between 425-475nm and 500-550nm, this probe is not conducive to use in the aqueous phase, and it is prone to fluorescence quenching in the presence of high concentrations of nitroreductase, and is mainly used in cell imaging in the biological field. Application potential of large-scale industrial enzymatic reactions

Chinese patent CN201710009923.7 discloses an AIE-based fluorescent probe for the detection of nitroreductase, which directly connects the nitro group to tetraphenylethylene, and before responding, due to the electronic effect of D-π-A, it makes The fluorescence is obvious. After the response, due to the D-π-D structure, the fluorescence becomes weaker and blue-shifted. The change in fluorescence is used to detect nitroreductase. However, this probe is mainly used in cells and cannot be applied to Detection and analysis of enzyme concentration in industrial enzymatic reaction system

[0007] Although fluorescent probes for the detection of nitroreductase have made some progress in the field of biological detection and imaging, they are rarely applied to the detection and analysis of enzyme activity in industrial enzymatic reactions.

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