Isolation, Identification and Characterization of Thermostable Amylase Producing Bacteria from Budamada Hot Spring, Misrak Badewacho, Hadiya Zone, Southern Ethiopia

Abstract

Thermophilic microorganisms are microorganisms that are stable over a wide range of harsh environmental conditions. Thermophillic amylase enzyme producing bacteria live at 45°C–80°C. Thermostable microbial amylases are more simplified and economical than other sources. The present research was aimed to isolate, identify and characterize thermostable amylase producing bacterial isolated from Budamada hot spring of Misrak Badawacho woreda. Thirty samples were collected from the source. Totally 18 isolates were isolated and 10 isolates were screened based on their ability to hydrolyze starch. Different biochemical tests were done to identify isolates based on Bergey’s Manual of systematic bacteriology. The results revealed that three enzymes from Bacillus spp. (SED2), Pseudomonas spp. (SED5) and Bacillus spp. (SED7) have maximum activity at 4% starch, 65˚C and pH of 8.5. Lactobacillus spp. (W20-0) and Bacillus cereus (SED12-1) at 8% starch, 75 and 65˚C, 8.5 and 7.5 pH respectively. Bacillus subtilis (W26-2) Bacillus licheniformis (W20-1), Lactobacillus spp. (SED10) at 6%starch; 35, 65, 45˚C; 6.5, 4.5, 4.5 PH., Bacillus spp.(SED8-3)&Bacillus spp. (W9) at 4% & 2%, 55˚C, 8.5 &4.5 pH and have maximum activity at 4% starch, 65˚C and pH of 8.5 and at 8% starch, 75 and 65˚C, 8.5 and 7.5 pH respectively., Lactobacillus spp. (SED10) at 6% starch; 35, 65, 45˚C; 6.5, 4.5, 4.5 PH. Bacillus spp. (SED8-3) and Bacillus spp.(W9) at 4% and 2%, 55˚C, 8.5 &4.5 PH. The amylase produced by bacterial isolates from Budamada hot spring was thermostable and performed from 65oC up to 75˚C. For most of the enzymes the optimum temperature was observed to be 65˚C; whereas optimum substrate concentration and pH for most of them were 4% starch and 8.5pH respectively. They were active at a various starch conditions (2%-10%), pH of 4.5-8.5, and temperature of 35-75°C.Generally, the bacteria isolated were thermophilic and produced thermostable amylase. Therefore, Budamada hot spring could be the source of thermophilic microorganisms and thermozymes as well.