Intestinal alkaline phosphatase bioautographic analysis of plant extracts

Abstract

Alkaline phosphatases are key enzymes in human physiology; they catalyze dephosphorylation reactions that are important in functions such as development of skeletal structure and metabolism. They are increasingly studied as targets for pharmacologic inhibition and thus molecules that inhibit Intestinal Alkaline Phosphatase (IAP) are becoming increasingly relevant. TLC bioautography is a low cost, relatively fast way of coupling complex mixture separation and qualitative bioassay analysis. Screening of complex mixtures involved applying a target mixture to the TLC plate, separating the mixture into components, and then spraying both enzyme and substrate onto a TLC plate. To initially test reaction conditions, napthyl phosphate with fast blue b are used as substrates to yield a blue azide product, and inhibition can therefore be qualitatively analyzed by a lack of colour on the TLC plate. A follow up experiment will subsequently measure IAP inhibition in various plant extracts via TLC bioautography. Extract samples displaying inhibition will be subsequently separated via TLC to examine the nature of inhibitors. Further investigation will involve examining the structural nature of these inhibitors.

Faculty Mentor: Dr. Tina Bott