Simulation and experimental study of fluorescence labeled polyphosphate in microthrix parvicella.

To study the binding mechanism of 4',6-diamidino-2-phenylindole (DAPI) and polyphosphate (Poly-P) and find fluorescent dyes which can dye Poly-P better, the interaction model of DAPI and Poly-P was calculated by the self-consistent-charge, density functional tight-binding (SCC-DFTB-D) method, and the binding sites of DAPI and Poly-P were analyzed. Further, Cy3, Rhodamine 6G and Fluorescein, which are structurally similar to DAPI, were selected to analyze their interactions with Poly-P. The binding energies and frontier orbital properties of the complexes were analyzed. These four fluorescent dyes were further used to dye the activated sludge smear and observe the fluorescence property. The Simulation results show that the N-containing indole ring structures in DAPI plays an important role in the interaction with Poly-P; the binding energies for DAPI, Cy3, Rhodamine 6G and Fluorescein with Poly-P are -42.6, -165.4, -34.7 and -28.9 kcal/mol, respectively. The frontier orbital properties for the complexes were studied, which further indicates that the interactions between Cy3, Rhodamine 6G and Poly-P are stronger than that of Fluorescein and Poly-P. The experimental results showed that Cy3 had excellent dyeing effect on Poly-P and could recognize them, while Fluorescein could not dye Poly-P. The experimental results were in good agreement with those predicted by simulation, which verified the correctness of our calculation method and provided a new strategy for finding more reliable, more sensitive and more economical fluorescent dyes capable of dyeing Poly-P.