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-丙酮酰基四氢蝶呤合成酶(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。中编码bPTPS-1的基因。 PCC 7942,生产BH4-葡萄糖苷。该突变体在BH4-葡萄糖苷生产中是正常的,表明bPTPS-I不参与体内BH4合成,并为我们带来了双模块多肽的新PTPS直向同源物(bPTPS-II)。在体外对重组Synechocooccus bPTPS-II进行了分析,结果表明PTPS活性高于人的酶。进一步的计算分析表明,分别在绿色硫细菌和蓝细菌中分别存在单和双模块bPTPS-II直系同源物,这对于BH4-糖苷的生产是众所周知的。总之,我们发现了新的细菌PTPS直系同源物,具有单域或双域结构并负责体内的BH4合成,从而揭示了所有细菌PTPS同源物。

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