双齿围沙蚕β-1,3-葡萄糖苷酶分离纯化及其酶学性质

于海慧; 宋淑梅; 佟长青; 李伟

doi:10.13386/j.issn1002-0306.2017.10.035

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

摘要

摘要: 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+、Mg²⁺、Fe²⁺、Ba²⁺、Ca²⁺对β-1,3-葡萄糖苷酶酶活力影响较小,Al³⁺、Cu²⁺、Zn²⁺、Ag⁺对β-1,3-葡萄糖苷酶酶活力抑制较大,其中Zn²⁺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。

Abstract: To isolate and purify β-1, 3-glucosidase from Perinereis aibuhitensis and study its properties, the crude β-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 filtration 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 k Da with SDS-PAGE. The optimal temperature for the β-1, 3-glucosidase was 50 ℃.The optimum p H was 7.The Kmand Vmaxof β-1, 3-glucosidase were8.2 × 10-4mol/L and 3.2 × 10-4μmol/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.