Production of thermostable extracellular α-amylase by a moderate thermophilic Bacillus licheniformis isolated from Qinarje Hot Spring (Ardebil prov. of Iran)

Abstract

Background and Purpose: Amylases are most important industrial enzymes that account for about 30% of the world’s food, feed, fermentation, textile, detergent and cellulosic industries. This study aimed at optimum production of thermostable α-amylase via moderate thermophilic bacterium (Bacillus licheniformis) which was recently isolated from Qinarje Hot spring.Material and Methods: Initially, ability of bacterium for amylase activity was determined by starch hydrolysis test using Gram’s iodine staining. Then bacterial growth pattern and amylase production curves in basal production medium were graphically determined at different time intervals. Finally, effect of different temperature, pH, carbon source, nitrogen source, minerals and inoculum size were studied on bacterial growth and amylase production using turbidimetric and DNS method, respectively.Results: Optimum enzyme production achieved after 84 hours of inoculation from cultures growing at 40 ˚C and pH 9.0 in a medium containing 0.03% (w/v) of CaCl2, compared to the basal medium, results showed that the best enzyme production happened with inoculum size of 4% (v/v). The addition of 1% (w/v) rice husk (as a Carbon source) enhanced enzyme productivity up to 160% and substitution of the peptone and yeast extract with 1% (w/v) of tryptone (as a Nitrogen source) increased the α-amylase production up to 160%.Conclusion: Our findings show that B. licheniformis-AZ2 strain has an ability to produce the thermostable α-amylase which is suitable in starch processing and food industries. To be commercialized, further investigation is required for enhancement of the enzyme production.Keywords: Bacillus licheniformis; Optimization; Basal medium; Agricultural by-products.