Cloning, Expression and Characterization of a Novel α-Amylasefrom Hot Springs of Northwestern Himalayas

Abstract

The present study utilized computational-guided experiments to conduct screening of a novel discovered acidic-thermostable α-amylase (AMYT) derived from a metagenome of hot springs. This approach was employed as a cost-effective alternative to the conventional method of functional screening. Initially, a computational screening methodology was employed to identify primary candidate that possess superior properties. AMYT was subjected to cloning, expression, purification, and characterization, among the candidates that were chosen. The AMYT enzyme demonstrated optimal activity at pH of 5.5 and a temperature of 50 °C. The enzyme demonstrated high efficacy in the presence of diverse chemicals, demonstrating a remarkable capacity in the hydrolysis of a wide range of substrates. Furthermore, it was found to be independent of Ca2+ ions. The results of this study demonstrated the efficacy of computational methods in identifying previously unknown acidic thermostable α-amylases. The accuracy of the selection method suggests that the AMYT has the potential to be a viable option for industrial starch processing, as it has the ability to enhance the output of final products and decrease overall costs.