6-Pyruvoyltetrahydropterin synthase (PTPS) catalyzes the second step of tetrahydrobiopterin (BH4) synthesis. We previously identified PTPS orthologs (bPTPS-Is) in bacteria which do not produce BH4. In this study we disrupted the gene encoding bPTPS-I in Synechococcus sp. PCC 7942, which produces BH4-glucoside. The mutant was normal in BH4-glucoside production, demonstrating that bPTPS-I does not participate in BH4 synthesis in vivo and bringing us a new PTPS ortholog (bPTPS-II) of a bimodular polypeptide. The recombinant Synechococcus bPTPS-II was assayed in vitro to show PTPS activity higher than human enzyme. Further computational analysis revealed the presence of mono and bimodular bPTPS-II orthologs mostly in green sulfur bacteria and cyanobacteria, respectively, which are well known for BH4-glycoside production. In summary we found new bacterial PTPS orthologs, having either a single or dual domain structure and being responsible for BH4 synthesis in vivo, thereby disclosing all the bacterial PTPS homologs.

译文

6-丙酮酸基四氢蝶呤合酶 (PTPS) 催化四氢生物蝶呤 (BH4) 合成的第二步。我们先前在不产生bh4的细菌中鉴定了PTPS直系同源物 (bPTPS-Is)。在这项研究中,我们破坏了Synechococcus sp. PCC 7942中编码bPTPS-I的基因,该基因产生了BH4-glucoside。该突变体在BH4-glucoside生产中是正常的,表明bPTPS-I在体内不参与BH4合成,并为我们带来了双峰多肽的新PTPS直系同源物 (bPTPS-II)。在体外测定了重组Synechococcus bptp-II,显示PTPS活性高于人酶。进一步的计算分析表明,分别在绿色硫细菌和蓝细菌中存在单和双模bPTPS-II直系同源物,这在BH4-glycoside生产中是众所周知的。总而言之,我们发现了新的细菌PTPS直系同源物,具有单个或双结构域结构,并负责体内BH4的合成,从而公开了所有细菌PTPS同源物。

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