ISOLATION AND CHARACTERIZATION OF AMYLASE-PRODUCING BACTERIA FROM DIFFERENT SOIL SAMPLES OF DISTRICT ABBOTTABAD

Abstract

Amylases are crucial hydrolase enzymes that can hydrolyze starch molecules and produce a variety of end-products like dextrin and small polymers made of glucose units. The current study addresses the lack of comprehensive knowledge about the diversity and potential of amylase-producing bacteria in different soil samples of Abbottabad district, Pakistan. The research seeks to isolate and characterize amylase-producing bacterial strains from six different locations, namely the Salhad damping site, Salhad fields, Havelian fruit mandi damping site, garden soil of Ayub Medical complex, Nathiagali, and Poultry farm Kakul village Abbottabad. Through this investigation, the study aims to identify promising amylase-producing bacterial isolates that can be utilized for potential biotechnological applications. The media used in the experiment included Luria- Bertani (LB) agar and broth, as well as starch agar media. The isolation of bacteria from the soil samples was done through serial dilution. Diluted samples were plated on LB agar and incubated, leading to the appearance of distinct colonies after 24 hours. Pure cultures were obtained from these colonies after additional incubation. The bacterial isolates were symmetrically labeled and then subjected to morphological identification using Gram staining and motility tests. Colony morphology was also observed by streaking bacteria on nutrient and LB media. Biochemical testing was performed, including the coagulase test, catalase test, oxidase test, indole test, and methyl-red test. For primary screening of amylase-producing bacteria, starch hydrolysis tests were conducted on starch agar media. Isolates showing significant zones of hydrolysis tests were selected for secondary screening. The potential isolates were assessed for their ability to produce amylase through submerged fermentation in a production medium. The crude enzyme was separated from the culture media and its activity was checked using starch agar plates. The study provides valuable insight into the diversity and potential of amylase-producing bacteria in Abbottabad district soil samples. The findings may have implications for further biotechnological applications, of enzyme production from these bacterial isolates and hold promise for cost-effective and eco-friendly alternatives in enzymes industries.