ISOLATION OF ORGANIC SOLVENT TOLERANT BACTERIA PRODUCING AMYLASE AND CHARACTERIZATION OF ITS AMYLASE ACTIVITY

Over the past few decades, the use of amylase enzymes to carry out reaction in organic media has achieved considerable interest in the research area of biotechnology research and development. Animal and plant sources of enzymes are unable to meet the industrial demand, and therefore, screening and purification of microbial organic solvent tolerant amylases are important. Organic solvent tolerant bacteria with an ability to produce extracellular organic solvent tolerant amylase were isolated form petrol contaminated area near Sewri, Mumbai, Maharashtra, India. These strains were screened for the amylolytic activity using starch agar plates and results indicated that it was amylase producer. Further, identification of bacterial strains was done using MALDI-TOF mass spectroscopy. The result of analysis indicated that all bacterial strains show highest degree of similarities with Bacillus spp. Moreover, the activity of organic solvent tolerant amylases was checked to study the effect of various parameters like temperature, pH and salt concentration to obtain optimum enzyme activity. The stability of the enzyme amylase in the presence of different solvents and metal ions was studied. Observed results indicate that enzyme activity was optimum between pH 6-8 and temperature range from 20o C to 70o C. Further the strains also show high relative activity in presence of Ca++ metal ions in the presence of various organic solvents with low logp activity. These characteristics of the enzyme in the presence of solvents are extremely favorable in lather industry, detergent industry and environmental remediation.