Purification and Chracterization of Ectracellular B Mannanase from a Thermophilic Bacterium, Geobacillus Stearothermophilus L-07  

Oleh:

Sumardi ; Suwanto, Antonius ; Suhartono, Maggy Thenawidjaja ; Purwadaria, Tresnawati

Jenis: Article from Journal - ilmiah nasional
Dalam koleksi: Jurnal Mikrobiologi Indonesia vol. 11 no. 2 (Sep. 2006), page 57-62.
Topik: Characterization; B mannanase; geobacillus strearothermophillus; thermophilic

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: JJ139 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

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Isi artikel

extracellular B mannanase (endo - 1,4 - B - D - mannanase, EC 3.2.1.78) from geobacillus stearothermophillus L - 07 grown on locust bean gum as a carbon source, was preciitated by ammonium sulfate. The enzyme complex was then purified through chromatographic columns including ion exchange (DEAE sephadex A - 50) and gel filtration (sephadex G - 100). Purification by DEAE sephadex A - 50 with concentration gradient of NaCl (0.2 , 0.3, and 0.4 M) separated the filtrate into three active fractions. The final gel filtration chromatohraphy on sephadex G - 100 from the second fraction yielded on major active fraction. The optimum pH after ammonium precipitation was 6.0 but it was shifted to 7.0 after purification. The optimum temperature were 80 and 70 Celcius degree for crude and purified enzyme respectively. The enzyme was not stable at 80 celcius degree and the activity could not be detected after 30 minutes incubation. Howeever it was stable at 70 celcius degree. The activity was decreasing in the course of incubation, but it still retained 57% of the total activity after 240 minutes. While addition of 1 M of NaCl retained the activity about 97% . At pH 4.0 -8.9 the enzyme retained aout 89% of their initial activities after 30 minuter incubation at 28 celcius degree. The fractons were collected and run on SDS poluacrylamide gel electrophoresis and the enzymes activity was detected following zymogram assay. Two b mannases of 73 kDa and 86 kDa were observed. The enzymes were endohydrolases that randomly hydrolyzed of B 1,4 - mannosidic linkages of galactomannan to mannootriose, mannobiose, and mannose.

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