Characteristics of thermostable chitinase enzymes from the indonesian Bacillus sp.13.26

A chitinase producing bacteria was screened and isolated from Tompaso hot spring in North Sulawesi Indonesia. The gram positive spore forming rod shaped bacteria was identified as Bacillus sp. 13.26 through sequence analysis of the 16SrRNA gene. When grown in medium containing 0.5% chitin at 55 °C, the thermophillic bacteria optimally produced extracellular chitinase enzyme after 72 h of incubation. The enzyme was purified through ammonium sulfate fractionation, dialysis, heat treatment at 60 °C for 3 h and chromatographed onto DEAE Sepharose CL-6B. The optimum temperature and pH of the 60-kD chitinase were found at 60 °C and pH 7–8. The thermostable chitinase still retained the activity after incubation for 5 h incubation at 70 °C. Zymogram analysis confirmed the thermostability of the enzyme ; when heated at 80 °C, the enzyme was still significantly active even after 1 h of incubation.