Production and Characterization of Chitinases from\ud Thermophilic Bacteria Isolated from Prataan Hot Spring,\ud East Java

摘要

Thermophilic bacteria producing chitinase were collected from Prataan hot spring, East Java. Indonesia and screened The isolated bacterium\udwas analyzed using 16S rRNA gene sequencing analysis and identified as\udPaenibacillus sp. The molecular identification was confirmed through\udmorphological and physiological analyses. The production of chitinase was\udconducted at various incubation times, temperatures, pH and concentrations of\udcolloidal chitin. The optimum condition of the isolate to produce the highest\udchitinase was 0.9% (w/v) of colloidal chitin (pH 7.0) at 48 °C for 24 hours. The\udobtained chitinases were optimally active at 55 oc and pH 6.0-7.0. The chitinases\udwere gradually purified by ammonium sulfate precipitation, Sephadex G-1 00 gel\udfiltration, followed by DEAE-cellulose ion exchange chromatography (IEC).\udThe purification method gave a purification factor of9.43 and a yield of2.68%.\udTwo protein fractions were obtained from sodium dodecyl sulfatepolyacrylamide\udgel electrophoresis (SDS-P AGE) with molecular weights of 68\udand82k.Da.