Isolation and Purification of Bacterial Psychrophilic Enantioselective Esterase for its potential use in pharmaceutical industry

Abstract

Enzymes have been used as biocatalyst since the very beginning of scientific knowledge. With advancement in research techniques and increase in awareness psychrophilic enzymes have attracted the attention of scientists. In this study a recently evolved Pseudomonal species has been isolated from Siachen glacier of Pakistan and its esterase producing capacity has been explored. The enzyme has been found to be active at 4oC, pH 10.5 both on Rhodamine B oil emulsion plates and UV spectroscopy using para nitro phenyl acetate at 410 nm. The concentration of the enzyme was found to be maximum in LB medium substituted with 1% yeast extract at 72 hours. Molecular weight determination through SDS-PAGE followed by MALDI-TOF mass spectrometry revealed the molecular mass 43 kDa. Our work has highlighted an industrially useful enzyme and its activity parameters have been optimized. Further purification will be done to determine the structure of the protein. The applications will be established practically. An alternative approach is to computationally predict the possible substrate of this enzyme and evaluate the outcomes experimentally.