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β-半乳糖苷和纤维二糖在 … 的交叉利用。

Cross-utilization of β-galactosides and cellobiose in .

机构信息

Department of Biotechnology and Food Engineering, Technion-Israel Institute of Technology, Haifa, Israel.

Schulich Faculty of Chemistry, Technion-Israel Institute of Technology, Haifa, Israel.

出版信息

J Biol Chem. 2020 Jul 31;295(31):10766-10780. doi: 10.1074/jbc.RA120.014029. Epub 2020 Jun 3.

Abstract

Strains of the Gram-positive, thermophilic bacterium possess elaborate systems for the utilization of hemicellulolytic polysaccharides, including xylan, arabinan, and galactan. These systems have been studied extensively in strains T-1 and T-6, representing microbial models for the utilization of soil polysaccharides, and many of their components have been characterized both biochemically and structurally. Here, we characterized routes by which utilizes mono- and disaccharides such as galactose, cellobiose, lactose, and galactosyl-glycerol. The genome encodes a phosphoenolpyruvate carbohydrate phosphotransferase system (PTS) for cellobiose. We found that the cellobiose-PTS system is induced by cellobiose and characterized the corresponding GH1 6-phospho-β-glucosidase, Cel1A. The bacterium also possesses two transport systems for galactose, a galactose-PTS system and an ABC galactose transporter. The ABC galactose transport system is regulated by a three-component sensing system. We observed that both systems, the sensor and the transporter, utilize galactose-binding proteins that also bind glucose with the same affinity. We hypothesize that this allows the cell to control the flux of galactose into the cell in the presence of glucose. Unexpectedly, we discovered that T-1 can also utilize lactose and galactosyl-glycerol via the cellobiose-PTS system together with a bifunctional 6-phospho-β-gal/glucosidase, Gan1D. Growth curves of strain T-1 growing in the presence of cellobiose, with either lactose or galactosyl-glycerol, revealed initially logarithmic growth on cellobiose and then linear growth supported by the additional sugars. We conclude that Gan1D allows the cell to utilize residual galactose-containing disaccharides, taking advantage of the promiscuity of the cellobiose-PTS system.

摘要

革兰氏阳性、嗜热细菌株 拥有精细的半纤维素多糖利用系统,包括木聚糖、阿拉伯聚糖和半乳糖聚糖。这些系统在代表土壤多糖利用的微生物模型菌株 T-1 和 T-6 中得到了广泛研究,其许多成分在生化和结构上都得到了表征。在这里,我们描述了 利用单糖和二糖(如半乳糖、纤维二糖、乳糖和半乳糖基甘油)的途径。 基因组编码了一个磷酸烯醇丙酮酸碳水化合物磷酸转移酶系统(PTS)用于纤维二糖。我们发现纤维二糖-PTS 系统被纤维二糖诱导,并表征了相应的 GH1 6-磷酸-β-葡萄糖苷酶 Cel1A。该细菌还拥有两种用于半乳糖的运输系统,一种是半乳糖-PTS 系统,另一种是 ABC 半乳糖转运体。ABC 半乳糖转运系统由一个三组分感应系统调节。我们观察到,这两个系统,即传感器和转运体,都利用与葡萄糖具有相同亲和力的半乳糖结合蛋白。我们假设这允许细胞在存在葡萄糖的情况下控制半乳糖流入细胞的通量。出乎意料的是,我们发现 T-1 还可以通过纤维二糖-PTS 系统与双功能 6-磷酸-β-半乳糖/葡萄糖苷酶 Gan1D 一起利用乳糖和半乳糖基甘油。在存在纤维二糖的情况下,菌株 T-1 的生长曲线,无论是用乳糖还是半乳糖基甘油,最初在纤维二糖上呈对数生长,然后在额外的糖的支持下呈线性生长。我们得出结论,Gan1D 允许细胞利用含有残余半乳糖的二糖,利用纤维二糖-PTS 系统的混杂性。

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