Navarro-Ródenas A, Morte A, Pérez-Gilabert M
Departamento de Biología Vegetal (Botánica), Facultad de Biología, Universidad de Murcia, Campus de Espinardo, Murcia, Spain.
Plant Biol (Stuttg). 2009 Sep;11(5):678-85. doi: 10.1111/j.1438-8677.2008.00172.x.
In the present paper, we confirmed that alkaline phosphatase (ALP) is the main phosphatase present in ascocarps of the edible mycorrhizal fungus Terfezia claveryi. The enzyme was partially purified by precipitation with polyethylene glycol. The purification achieved from a crude extract was fivefold, with 53% of the activity recovered, and acid phosphatase, most of the lipids and phenolic compounds were eliminated. Alkaline phosphatase was kinetically characterised at pH 10.0, the optimum for this enzyme, using p-nitrophenyl phosphate as substrate. The V(max) and K(m) values were 0.3 micromol.min(-1).mg(-1) protein and 9.0 mM, respectively. Orthovanadate was a competitive inhibitor of ALP, with a K(i) of 42.5 microM. The enzyme was histochemically localised in the peridium, the hypothecium and in the ascogenic hyphae of the gleba using both colour and fluorescent reactions. The results presented suggest that the ascocarp of T. claveryi, at some stages of its development, may become nutritionally autonomous and independent of the host plant.
在本论文中，我们证实碱性磷酸酶（ALP）是可食用菌根真菌克拉氏块菌子囊果中存在的主要磷酸酶。该酶通过聚乙二醇沉淀进行了部分纯化。从粗提物中实现了五倍的纯化，回收了53%的活性，并且消除了酸性磷酸酶、大部分脂质和酚类化合物。在pH 10.0（该酶的最适pH）下，以对硝基苯磷酸为底物对碱性磷酸酶进行了动力学表征。V(max)和K(m)值分别为0.3微摩尔·分钟(-1)·毫克(-1)蛋白质和9.0毫摩尔。原钒酸盐是ALP的竞争性抑制剂，K(i)为42.5微摩尔。使用显色和荧光反应，该酶在组织化学上定位于包被、子实下层和子实层的产囊菌丝中。所呈现的结果表明，克拉氏块菌的子囊果在其发育的某些阶段可能在营养上自主且独立于寄主植物。
