Amylase enzyme production and investigation of possibilities in industrial usage from thermophile moderately halophilic Bacillus sp

In this study, haloalkalophile, thermophile and thermostable amylase enzyme from Bacillus sp strains was isolated and characterised. Therefore the temperature and pH range was determined for enzyme production and bacterial growth on solid media. Optimum activity temperature, optimum pH, thermal stability, pH stability of the isolated amylase as well as effect of NaCl, some inhibitors, chelators, metal ions and detergents on enzyme activity were determined. SDS-PAGE analyses were also carried out. Analyses of the enzyme by SDSPAGE revealed two independent bands (116.000 and 97400 Da.). While the bacteria grow at between 20-60OC, the enzyme production was occurred between 20-55oC. But the highest growth rate and enzyme production were obtained 25-55oC with an average %84. As isolated strains showed growth and enzyme synthesis at pH 7.0-12.5 range, optimum growth and enzyme production occurred at pH 8.5 and 55oC. Among the strains, SB-28 showed the maximum enzyme synthesis at pH 9.5 and 55oC. Optimum activity of the SB-28 amylase was at pH12.5 and 140oC. The enzyme was retained it`s activity around %54 at the end of 72 hours between pH 9.0 and 12.0. The enzyme after 120 min preincubation at 140oC was not inactivated, instead activated to be around 408%. On the other hand, the enzyme showed the maximum activity, 89%, on 10% NaCl concentration after 60 min incubation at 55oC. Additionally, SB-28 retained it`s activity in the presence of EDTA, CaCl2, PMSF, 2-Mercaptoetanol, SDS, Triton-X, Na-Sulfite, NaCl, ZnCl2, 1-10Phenantroline and the urea at the rate of 65.42%, 70.20%, 74.23%, 81.0%,71.0%, 82.03%,80.0%,74.20%, 68.10%,65.40% and11.20% respectively. According to these results, the isolated enzyme is resistand to detergents, and halophile, extreme alkalophile, pH stable, ultrathermophile, and inducible thermostable. As a result this SB- 28 amylase enzyme could find an important place for the usage in textile and starch liquefaction industries.