双齿围沙蚕β-1,3-葡萄糖苷酶分离纯化及其酶学性质

Purification and characterization ofβ-1,3-glucosidase from Perinereis aibuhitensis

从双齿围沙蚕中提取、分离纯化β-1,3-葡萄糖苷酶,并研究其酶学性质。通过80%饱和度(NH4)2SO4沉淀从双齿围沙蚕中得到粗β-1,3-葡萄糖苷酶,粗酶依次经DEAE-52离子交换层析、Sephadex G-100凝胶过滤层析进行分离纯化。酶纯化倍数为35.74,回收率为39.49%。SDS-PAGE检测表明其分子量为28.7 k Da。该酶最适温度为50℃,最适p H为7,Km为8.2×10-4mol/L,Vmax为3.2×10-4μmol/h。金属离子K+、Mg^(2+)、Fe^(2+)、Ba^(2+)、Ca^(2+)对β-1,3-葡萄糖苷酶酶活力影响较小,Al^(3+)、Cu^(2+)、Zn^(2+)、Ag^+对β-1,3-葡萄糖苷酶酶活力抑制较大,其中Zn^(2+)抑制作用最强。双齿围沙蚕可作为β-1,3-葡萄糖苷酶的潜在来源。

To isolate and purifyβ-1,3 - glucosidase from Perinereis aibuhitensis and study its properties, the crudeβ-1,3 - glucosidase was obtained from the P.aibuhitensis by ammonium sulfate precipitation, which was purified using ion-exchange chromatography on DEAE- 52 and gel fihration chromatography on Sephadex G- 100. Finally the β- 1,3 - glucosidase was purified 35.74-fold and 39.49% recovery yield.The β-1,3-glucosidase was estimated to be 28.7 kDa with SDS-PAGE.The optimal temperature for the ,8-1,3-glucosidase was 50 ℃.The optimum pH was 7.The Km and Vmax offl-1,3-glucosidase were 8.2 ×10^-4mol/L and 3.2 × 10^-4 μmol/h, respectively.The enzyme activity was not affected by K＋ , Mg2＋ , Fe2＋ , Ba2＋ and Ca2＋ ,while inactivated by Al3＋, Cu2＋, Zn2＋ and Ag＋, and especially strongly inactivated by Zn2＋ .P.aibuhitensis might be a candidate resource in producing β- 1,3- glucosidase.