El-Gindy A A, Saad R R, Fawzi E
Dept. of Biological Sciences, Fac. of Education, Cairo, Egypt.
Acta Microbiol Pol. 2003;52(1):35-44.
Exo-1,4-beta-glucanase (E.C. 3.2.1.91) was successively purified by precipitation with acetone, followed by gel filtration on Sephadex G-100 and chromatographed onto DEAE-cellulose. A typical procedure provided 47.14 fold purification with 72.8% yield. The molecular mass of the purified enzyme was found to be 88 kDa by SDS-PAGE. The pH optimum of the enzyme was 5.2 and maximum activity was obtained at 45 degrees C. Km value against alpha-cellulose was 0.65 mg mL(-1). Alpha-cellulose and filter paper were the best substrates for enzyme activity. Enzyme was activated by Mn2+ and Fe3+, inactivated by Cu2+ and completely inhibited by Hg2+ and Ag+.
外切-1,4-β-葡聚糖酶（E.C. 3.2.1.91）先后通过丙酮沉淀、Sephadex G-100凝胶过滤和DEAE-纤维素柱层析进行纯化。一个典型的流程可实现47.14倍的纯化，产率为72.8%。通过SDS-PAGE测定，纯化酶的分子量为88 kDa。该酶的最适pH为5.2，在45℃时活性最高。以α-纤维素为底物时的Km值为0.65 mg mL(-1)。α-纤维素和滤纸是该酶活性的最佳底物。该酶被Mn2+和Fe3+激活，被Cu2+失活，被Hg2+和Ag+完全抑制。
