Abstract
A new endo-$\beta$-1, 4-glucanase was purified from the culture filtrate of thermophilic anaerobic Clostridium thermocellum. The purification procedure included two steps of ion exchange chromatography with DEAD-Sephadex A-50 and gel filtration chromatography with Sephadex G-75. Even though the 56 fold increase in CMCase specific activity was obtained, the actually recovered enzyme activity was relatively lower level of 0.7%. Judging from the two bands in SDS-polyacrylamide gel electrophoresis, the endo-$\beta$-1, 4-glucanase consists of two subunits whose M.W. are 38,000 and 58,000, respectively. The optimum pH and temperature were determined to be 5.0 and $65^{\circ}C$, respectively. The enzyme was stable up to $70^{\circ}C$, but inactivated at $80^{\circ}C$. The kinetic parameters of the separated fraction were also determined. The purified enzyme did not show any significant hydrolytic activity against the highly ordered crystalline cellulose as well as filter paper.
고온 혐기성 Clostridium thermocellum의 배양액으로부터 새로운 endo-$\beta$-1, 4-glucanase를 ion exchange chromatography와 gel filtration chromatongraphy를 통하여 정제하였다. 정제된 효소의 비활성은 56배 증가하였으나 수율은 0.7%로서 매우 낮았다. SDS-PAGE, 결과, 정제된 효소는 분자량이 각각 38,000과 58,000으로 된 두 개의 subunit로 구성되어 있었다. 이 효소의 반응 최적 pH는 5.0 최적온도는 $65^{\circ}C$였으며 $70^{\circ}C$까지는 열에 안정하였으나 $80^{\circ}C$에서 거의 실활되었다. 기타 여러 가지 효소학적인 성질을 조사하였으며, 분리된 효소는 결정형 섬유소에 대한 효소활성을 나타내지 않았다.