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토양으로부터 분리한 Bacillus agaradhaerens DK-2386 균주가 생산하는 Xylanase의 특성

Characterization of Xylanase from Bacillus agaradhaerens DK-2386 Isolated from Korean Soil

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주제분류생물학

초록

Bacillus agaradhaerens 균주의 효소를 정제하여 그 특성을 확인하였다. 균주가 생산하는 xylanase를 70% ammonium sulfate로 침전하고 CM-sepharose ion exchange chromato-graphy와 ultrafiltration(PM-10)을 행하여 16.7%의 수율과 6.7배의 정제도를 지니는 효소를 얻을 수 있었으며 SDSPAGE를통하여 23 kDa의 분자량을 갖는 효소임을 확인하였다. 효소의 최적 pH는 6.0으로서 sodium phosphate buffer에서 24시간까지 안정하였으며, 최적반응온도는 60℃, EDTA의 첨가 시 효소활성이 증가되었고 40℃에서 24시간까지 안정하였다. 효소는 xylan만을 기질로 이용할 수 있었으며 Birchwood xylan에 대하여 최대 활성을 나타내었고 Vmax는 49,724 μM/min, Km은 6.08 mg/ml로 확인되었다.

Abstract

The optimum conditions for the production of xylanase from Bacillus agaradhaerens DK-2386 have been previously investigated. In this study xylanase was purified by ammonium sulfate precipitation and CM-sepharose ion exchange chromatography. The molecular mass of the xylanase as determined by SDS-PAGE was 23 kDa in a form of monomeric enzyme. The optimum pH and temperature for xylanase activity was 6.0 and 60℃, respectively. Xylanase activity was increased by the addition of EDTA and then stabilized at 40℃ for 24 h. The maximum xylanase activity was obtained when Birchwood xylan was used as a substrate and the Vmax and Km were 49,724 μmol/min and 6.08 mg/ml, respectively.

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