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首页> 外文期刊>Journal of bacteriology >Glucosylglycerate Biosynthesis in the Deepest Lineage of the Bacteria: Characterization of the Thermophilic Proteins GpgS and GpgP from Persephonella marina
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Glucosylglycerate Biosynthesis in the Deepest Lineage of the Bacteria: Characterization of the Thermophilic Proteins GpgS and GpgP from Persephonella marina

机译：细菌最深谱系中的葡萄糖基甘油酸酯生物合成：滨海伯生不动杆菌嗜热蛋白GpgS和GpgP的表征

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The pathway for the synthesis of glucosylglycerate (GG) in the thermophilic bacterium Persephonella marina is proposed based on the activities of recombinant glucosyl-3-phosphoglycerate (GPG) synthase (GpgS) and glucosyl-3-phosphoglycerate phosphatase (GpgP). The sequences of gpgS and gpgP from the cold-adapted bacterium Methanococcoides burtonii were used to identify the homologues in the genome of P. marina, which were separately cloned and overexpressed as His-tagged proteins in Escherichia coli. The recombinant GpgS protein of P. marina, unlike the homologue from M. burtonii, which was specific for GDP-glucose, catalyzed the synthesis of GPG from UDP-glucose, GDP-glucose, ADP-glucose, and TDP-glucose (in order of decreasing efficiency) and from d-3-phosphoglycerate, with maximal activity at 90°C. The recombinant GpgP protein, like the M. burtonii homologue, dephosphorylated GPG and mannosyl-3-phosphoglycerate (MPG) to GG and mannosylglycerate, respectively, yet at high temperatures the hydrolysis of GPG was more efficient than that of MPG. Gel filtration indicates that GpgS is a dimeric protein, while GpgP is monomeric. This is the first characterization of genes and enzymes for the synthesis of GG in a thermophile.
机译：基于重组葡萄糖基-3-磷酸甘油酸合酶（GPGS）和葡萄糖基-3-磷酸甘油酸磷酸酶的活性，提出了嗜热细菌 Persephonella marina 中葡萄糖基甘油酸（GG）的合成途径。 （GpgP）。用冷适应细菌 Burthanii 的 gpgS 和 gpgP 序列鉴定 P基因组中的同源物。玛丽娜，分别克隆并在大肠杆菌中表达为带有His标签的蛋白。 P的重组GpgS蛋白。码头，与 M的同系物不同。专用于GDP葡萄糖的burtonii 催化UDP-葡萄糖，GDP-葡萄糖，ADP-葡萄糖和TDP-葡萄糖（按降序排列）和d-3-磷酸甘油酸酯合成GPG ，在90°C时具有最大活性。重组GpgP蛋白，如 M。 burtonii的同系物，去磷酸化的GPG和3-磷酸甘露糖基甘油酸酯（MPG）分别为GG和甘露糖基甘油酸酯，但是在高温下，GPG的水解效率要比MPG高。凝胶过滤表明GpgS是二聚体蛋白，而GpgP是单体。这是嗜热菌中GG合成的基因和酶的首次表征。

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