Fluorogenic Peptide Substrate for Quantification of Bacterial Enzyme Activities.

摘要: A novel peptide substrate (A G P L G) was developed for quantifying the activities of bacterial enzymes using a highly sensitive Fluorescence Resonance Energy Transfer (FRET) based assay. The cleaved by collagenase class I, II, Liberase MTF C/T, NB1, and thermolysin/neutral protease, which significantly enhanced in presence CaCl2. However, these were decreased ZnSO4 or ZnCl2. Collagenase protease share similar cleavage sites, L↓G P↓G. NB1 cleaves at G↓P P↓L, addition to enzyme activity is pH dependent, within range 6.8 7.5, but diminished 8.0. Interestingly, not endogenous pancreatic such as trypsin, chymotrypsin, elastase. In conclusion, collagenase, specific can be applied quantify from different microbes. Furthermore, assay used fine-tuning reaction mixtures various agents enhance overall cocktail multiple achieve optimal organ/tissue digestion, while protecting integrity target cells.