A FAST FLUORESCENT PROBES OF TRYPSIN BASED ON PYRENE DERIVATIVES IN AQUEOUS SOLUTION

摘要

Trypsin is the most important digestive enzymes in the intestinal of human and animal , abnormal levels of trypsin will result in many diseases,such as pancreatitis, pancreatic cancer. Therefor rapid and sensitive detection of trypsin is very improtant and significant. Fluorescent probes are widely used in the field of environmental sciences and life sciences because of its high sensitivity, high selectivity, in-situ real-time fast, non-destructive testing of samples, etc.. As one of the most important fluorophore, the aggregation of pyrene monomer molecules (emission at 370-420 nm)could produce excimer emission. As a result, a red-shiftedand broad emission band at 450-550 nm could be observed. In this work, l-hydroxy-3-pyrene (pyrene-1-oxy) propyl sulfonate (Pypos) was synthesized using the 1-hydroxy-pyrene as a raw material. In aqueous solution, Pypos mainly exists in the monomeric form. When Protamine used as substrate is added, strong attractive electrostatic and hydrophobic interactions between Protamine and Pypos result in the aggregation of Pypos,a red-shifted at 485nm could be observed. Upon the addition of Trypsin,the Protamine is hydrolyzed leading to the deaggregation of Protamine, the pyrene excimer returns to the monomeric state.This excimer-monomer transition afford a convenient fluorometric assay for Trypsin,and the detection limit is set to 0.5ug/mL.