2株茶藨生柱锈重寄生木霉胞壁降解酶的基本性质研究

The Study on Degrading Enzymes of Cell Wall from Two Mycoparasites of Cronartium ribicola

为了给β-1,3葡聚糖酶和几丁质酶的分离纯化及重寄生菌的利用提供理论依据,从茶藨生柱锈（Cronartium ribicola J.C.Fischer）的重寄生菌TR1和TR2中提取了β-1,3葡聚糖酶和几丁质酶,并进行了粗酶活性测定。结果显示,2菌株所产β-1,3-葡聚糖酶最适温度均为35℃,TR1所产β-1,3葡聚糖酶的最适pH4.0,Km值为43.82μg/mL,Ca^2＋是其激活剂,Cu^2＋、Fe^3＋、Al^3＋、Zn^2＋为其抑制剂。TR2所产β-1,3葡聚糖酶的最适pH5.0,Km为71.28μg/mL,Ca^2＋和Mn^2＋是其激活剂,Cu^2＋、Fe^3＋、Al^3＋为其抑制剂。2菌株所产几丁质酶的激活剂为Mn2＋和Ca2＋,抑制剂为Cu^2＋。TR1所产几丁质酶的最适温度为40℃,最适pH6.0,Km为92.49μg/mL;TR2所产几丁质酶的最适温度范围较宽为30~50℃,最适pH5.0,Km为71.11μg/mL。TR1和TR2所产β-1,3-葡聚糖酶和几丁质酶稳定性较高,直接利用粗酶在野外防治疱锈病具有一定的可行性。

The aim was to provide the basis for the isolation and purification of β-1,3-glucanase and chitinase, and the use of mycoparasite. β-1,3-glucanase and chitinase were isolated from two mycoparasites （TR1 and TR2） of Cronartium ribicola. The enzymatic activities of β-1,3-glucanase and chitinase were detected. The results showed that optimum temperature of β-1,3-glucanase from TR1 and TR2 was 35℃. The optimum pH of β-1,3-glucanase from TR1 was 4.0 and Km was 43.82 μg/mL. Ca^2＋ was activator of the enzyme and Cu^2＋, Fe^3 ＋ , Al^3 ＋ , Zn^2 ＋ were inhibitors. The optimum pH of β-1,3-glucanase from TR2 was 5.0 and Km was 71.28 μg/mL. Ca2＋ and Mn^2＋ were activators of the enzyme and Cu^2＋, Fe^3＋, Al^3＋ were inhibitors. Activators of chitinase from TR1 and TR2 were Mn2＋ and Ca^2＋ and inhibitor was Cu^2＋. The optimum temperature of chitinase from TR1 was 40℃ and the optimum pH was 6.0, Km of the enzyme was 92.49 μg/mL. The optimum temperature of chitinase from TR2 was from 30℃ to 50℃. The optimum pH was 5.0 and Km was 71.11 μg/mL. β-1,3-glucanase and chitinase produced by TR1 and TR2 possess high stability, so that it was feasible to use crude enzyme directly in field control of blister rust.