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肿胀素是一种里氏木霉蛋白,其序列与植物扩张蛋白相似,对纤维素材料具有分解活性。

Swollenin, a Trichoderma reesei protein with sequence similarity to the plant expansins, exhibits disruption activity on cellulosic materials.

作者信息

Saloheimo Markku, Paloheimo Marja, Hakola Satu, Pere Jaakko, Swanson Barbara, Nyyssönen Eini, Bhatia Amit, Ward Michael, Penttilä Merja

机构信息

VTT Biotechnology, PO Box 1500, 02044 VTT, Finland.

出版信息

Eur J Biochem. 2002 Sep;269(17):4202-11. doi: 10.1046/j.1432-1033.2002.03095.x.

Abstract

Plant cell wall proteins called expansins are thought to disrupt hydrogen bonding between cell wall polysaccharides without hydrolyzing them. We describe here a novel gene with sequence similarity to plant expansins, isolated from the cellulolytic fungus Trichoderma reesei. The protein named swollenin has an N-terminal fungal type cellulose binding domain connected by a linker region to the expansin-like domain. The protein also contains regions similar to mammalian fibronectin type III repeats, found for the first time in a fungal protein. The swollenin gene is regulated in a largely similar manner as the T. reesei cellulase genes. The biological role of SWOI was studied by disrupting the swo1 gene from T. reesei. The disruption had no apparent effect on the growth rate on glucose or on different cellulosic carbon sources. Non-stringent Southern hybridization of Trichoderma genomic DNA with swo1 showed the presence of other swollenin-like genes, which could substitute for the loss of SWOI in the disruptant. The swollenin gene was expressed in yeast and Aspergillus niger var. awamori. Activity assays on cotton fibers and filter paper were performed with concentrated SWOI-containing yeast supernatant that disrupted the structure of the cotton fibers without detectable formation of reducing sugars. It also weakened filter paper as assayed by an extensometer. The SWOI protein was purified from A. niger var. awamori culture supernatant and used in an activity assay with Valonia cell walls. It disrupted the structure of the cell walls without producing detectable amounts of reducing sugars.

摘要

被称为扩展蛋白的植物细胞壁蛋白被认为可破坏细胞壁多糖之间的氢键而不水解它们。我们在此描述了一个与植物扩展蛋白具有序列相似性的新基因,它是从纤维素分解真菌里氏木霉中分离出来的。名为肿胀素的这种蛋白质具有一个N端真菌型纤维素结合结构域,该结构域通过一个连接区与类扩展蛋白结构域相连。该蛋白质还含有与哺乳动物纤连蛋白III型重复序列相似的区域,这是首次在真菌蛋白中发现。肿胀素基因的调控方式与里氏木霉纤维素酶基因大体相似。通过破坏里氏木霉的swo1基因来研究SWOI的生物学作用。这种破坏对在葡萄糖或不同纤维素碳源上的生长速率没有明显影响。用swo1对木霉基因组DNA进行非严格的Southern杂交,结果表明存在其他类似肿胀素的基因,它们可以替代破坏菌株中SWOI的缺失。肿胀素基因在酵母和黑曲霉变种泡盛曲霉中表达。用含有浓缩SWOI的酵母上清液对棉纤维和滤纸进行活性测定,该上清液破坏了棉纤维的结构,但未检测到还原糖的形成。通过引伸计测定,它还使滤纸变弱。从黑曲霉变种泡盛曲霉培养上清液中纯化出SWOI蛋白,并用于对瓦罗尼亚细胞壁的活性测定。它破坏了细胞壁的结构,但未产生可检测量的还原糖。

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