花生几丁质酶的纯化及酶学性质研究

Purification and Enzymatic Characterization of the Chitinase from Arachis hpyogaea

本课题利用亲和色谱和凝胶过滤色谱法从花生种子中分离得到了几丁质酶.经SDS-PAGE鉴定,该酶蛋白已达电泳纯,分子量约34.4 kD.在还原和非还原状态下均显示单一区带,表明其为单体蛋白.此外,通过等电聚焦法可测得该酶等电点为5.1.而酶学性质的相关研究则表明:该酶最适pH为5.4,最适反应温度为40～50℃.

A chitinase was purified from seeds of peanut （Arachis hpyogaea） by aqueous extraction, affinity chromatography （Affi-gel） and Gel-filtration chromatography （Sephadex G-75）. The purified enzyme showed a single band on SDS-PAGE with the molecular weight about 34.4 kD in both reduced and non-reduced conditions, indicating that the enzyme is a monomeric protein. The pI was measured to be 5.1 by isoelectric focusing electrophoresis. The enzyme exhibited its optimum activity at pH 5.4, and a temperature between 40℃ and 50℃.