School of Agriculture, Utsunomiya University, 350 Mine-machi, Utsunomiya, Tochigi, 321-8505, Japan; United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan.
School of Agriculture, Utsunomiya University, 350 Mine-machi, Utsunomiya, Tochigi, 321-8505, Japan; Center for Bioscience Research and Education, Utsunomiya University, 350 Mine-machi, Utsunomiya, Tochigi, 321-8505, Japan.
Protein Expr Purif. 2020 Jun;170:105609. doi: 10.1016/j.pep.2020.105609. Epub 2020 Feb 16.
Brown-rot fungi preferentially degrade softwood and cause severe breakdown of wooden structures. At the initial stage of the brown-rot decay, penetrating hyphae of the fungi are observed in ray parenchyma. Since starch grains are known to be present in the ray parenchyma of sapwood, investigation of the functions and roles of the starch-degrading enzymes is important to understand the initial stage of brown-rot decay. We purified and characterized two starch-degrading enzymes, an α-amylase (FpAmy13A) and a glucoamylase (FpGLA15A), from the brown-rot fungus, Fomitopsis palustris, and cloned the corresponding genes. The optimal temperature for both enzymes was 60 °C. FpAmy13A showed higher activity at a broad range of pH from 2.0 to 5.0, whereas FpGLA15A was most active at pH 5.0-6.0. Notable thermal stability was found for FpGLA15A. Approximately 25% of the activity remained even after treatment at 100 °C for 30 min in sodium phosphate buffer at pH 7.0. These different characteristics imply the different roles of these enzymes in the starch degradation of wood.
褐腐真菌优先降解软木,导致木质结构严重崩溃。在褐腐腐朽的初始阶段,可以观察到真菌穿透射线薄壁组织的菌丝。由于众所周知,边材射线薄壁组织中存在淀粉颗粒,因此研究淀粉降解酶的功能和作用对于了解褐腐腐朽的初始阶段很重要。我们从褐腐菌 Fomitopsis palustris 中纯化和表征了两种淀粉降解酶,一种α-淀粉酶(FpAmy13A)和一种糖化酶(FpGLA15A),并克隆了相应的基因。这两种酶的最佳温度均为 60°C。FpAmy13A 在 pH 值为 2.0 到 5.0 的较宽范围内表现出更高的活性,而 FpGLA15A 在 pH 值为 5.0 到 6.0 时最活跃。发现 FpGLA15A 具有显著的热稳定性。在 pH 7.0 的磷酸钠缓冲液中,100°C 处理 30 分钟后,其活性仍保留约 25%。这些不同的特性表明这些酶在木材淀粉降解中具有不同的作用。