Rapid screening of enzyme inhibitors using Profiling of Enzyme-Metabolite Assay by HPLC (PREMA-HPLC)

Abstract

A number of isolates from different ecosystems were screened for their ability to inhibit tyrosinase resulting in the selection of isolate CFR 101, which showed an inhibition of 72%. The metabolites present in the crude extract of the selected isolate was profiled through high-performance liquid chromatography (HPLC) before the enzyme inhibition assay to reveal a 66% decrease in area of the peak at room temperature for 13.9 min, after the assay. Upon purification, this peak was identified as kojic acid, a known inhibitor of tyrosinase. This unique technique of combining a reaction assay mixture with HPLC profile wherein inhibitors can be rapidly pinpointed in crude extracts addresses the drawback of rapid chemical high-throughput screening (HTS) systems, which is limited to the chemical nature of metabolites without any evidence of their biological activities.

Keywords::

• Tyrosinase inhibitors
• HPLC
• screening
• kojic acid

Acknowledgements

The funding for this work from the 11th Five Year Plan Network Project on “Exploitation of India’s rich microbial diversity” is gratefully acknowledged. The assistance of Mr. Dasagrandhi Chakradhar in the early part of this work is acknowledged.

Declaration of interest

The funding for this work from the 11th Five Year Plan Network Project on “Exploitation of India’s rich microbial diversity” is gratefully acknowledged.