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来自沼泽腐朽菌的草酸盐外排转运蛋白。

Oxalate efflux transporter from the brown rot fungus Fomitopsis palustris.

机构信息

Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.

出版信息

Appl Environ Microbiol. 2010 Dec;76(23):7683-90. doi: 10.1128/AEM.00829-10. Epub 2010 Oct 1.

Abstract

An oxalate-fermenting brown rot fungus, Fomitopsis palustris, secretes large amounts of oxalic acid during wood decay. Secretion of oxalic acid is indispensable for the degradation of wood cell walls, but almost nothing is known about the transport mechanism by which oxalic acid is secreted from F. palustris hyphal cells. We characterized the mechanism for oxalate transport using membrane vesicles of F. palustris. Oxalate transport in F. palustris was ATP dependent and was strongly inhibited by several inhibitors, such as valinomycin and NH(4)(+), suggesting the presence of a secondary oxalate transporter in this fungus. We then isolated a cDNA, FpOAR (Fomitopsis palustris oxalic acid resistance), from F. palustris by functional screening of yeast transformants with cDNAs grown on oxalic acid-containing plates. FpOAR is predicted to be a membrane protein that possesses six transmembrane domains but shows no similarity with known oxalate transporters. The yeast transformant possessing FpOAR (FpOAR-transformant) acquired resistance to oxalic acid and contained less oxalate than the control transformant. Biochemical analyses using membrane vesicles of the FpOAR-transformant showed that the oxalate transport property of FpOAR was consistent with that observed in membrane vesicles of F. palustris. The quantity of FpOAR transcripts was correlated with increasing oxalic acid accumulation in the culture medium and was induced when exogenous oxalate was added to the medium. These results strongly suggest that FpOAR plays an important role in wood decay by acting as a secondary transporter responsible for secretion of oxalate by F. palustris.

摘要

一种草酸发酵褐腐真菌,木蹄层孔菌,在木材腐烂过程中会大量分泌草酸。草酸的分泌对于木质细胞壁的降解是必不可少的,但对于草酸从木蹄层孔菌丝状细胞中分泌的运输机制几乎一无所知。我们使用木蹄层孔菌的膜泡来描述草酸运输的机制。木蹄层孔菌中的草酸运输依赖于 ATP,并且被几种抑制剂(如缬氨霉素和 NH4+)强烈抑制,这表明该真菌中存在一种次级草酸转运蛋白。然后,我们通过在含有草酸的平板上培养酵母转化子的功能筛选,从木蹄层孔菌中分离出一个 cDNA,FpOAR(木蹄层孔菌草酸抗性)。FpOAR 预测是一种膜蛋白,具有六个跨膜结构域,但与已知的草酸转运蛋白没有相似性。具有 FpOAR(FpOAR 转化子)的酵母转化子对草酸具有抗性,并且比对照转化子含有更少的草酸。使用 FpOAR 转化子的膜泡进行的生化分析表明,FpOAR 的草酸转运特性与在木蹄层孔菌的膜泡中观察到的特性一致。FpOAR 转录本的数量与培养基中草酸积累量的增加相关,并在外源草酸添加到培养基中时被诱导。这些结果强烈表明,FpOAR 通过作为负责木蹄层孔菌草酸分泌的次级转运蛋白,在木材腐烂中发挥重要作用。

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