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Biosynthesis of an Opine Metallophore by Pseudomonas aeruginosa
Biochemistry ( IF 2.9 ) Pub Date : 2017-11-03 00:00:00 , DOI: 10.1021/acs.biochem.7b00804 Jeffrey S. McFarlane 1 , Audrey L. Lamb 1
Biochemistry ( IF 2.9 ) Pub Date : 2017-11-03 00:00:00 , DOI: 10.1021/acs.biochem.7b00804 Jeffrey S. McFarlane 1 , Audrey L. Lamb 1
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Bacterial pathogenesis frequently requires metal acquisition by specialized, small-molecule metallophores. We hypothesized that the Gram-negative Pseudomonas aeruginosa encodes the enzymes nicotianamine synthase (NAS) and opine dehydrogenase (ODH), biosynthesizing a new class of opine metallophore, previously characterized only in the unrelated Gram-positive organism Staphylococcus aureus. The identity of this metallophore, herein named pseudopaline, was determined through measurements of binding affinity, the in vitro reconstitution of the biosynthetic pathway to screen potential substrates, and the confirmation of product formation by mass spectrometry. Pseudopaline and the S. aureus metallophore staphylopine exhibit opposite stereochemistry for the histidine moiety, indicating unique recognition by NAS. Additionally, we demonstrate SaODH catalysis in the presence of pyruvate, as previously shown, but also oxaloacetate, suggesting the potential for the production of a variant form of staphylopine, while PaODH specifically recognizes α-ketoglutarate. Both the staphylopine and pseudopaline operons have been implicated in the pathogenesis of key infectious disease states and warrant further study.
中文翻译：
铜绿假单胞菌对高山金属团的生物合成
细菌的发病机理经常需要通过专门的小分子金属团获得金属。我们假设革兰氏阴性铜绿假单胞菌编码烟碱胺合酶（NAS）和鸦片脱氢酶（ODH）酶，生物合成了一类新型的鸦片金属荧光团，以前仅在无关的革兰氏阳性生物体金黄色葡萄球菌中有特征。通过测量结合亲和力，体外合成筛选潜在底物的生物合成途径以及通过质谱法确认产物形成，来确定该金属团簇的身份，在本文中称为假papalpaline 。伪帕林和金黄色葡萄球菌金属团葡萄球碱对组氨酸部分显示相反的立体化学，表明被NAS唯一识别。此外，我们证明了丙酮酸存在下的SaODH催化作用（如先前所示），草酸乙酸酯也存在，这表明了产生葡萄球菌素变体形式的潜力，而PaODH特异性识别α-酮戊二酸。葡萄球菌操纵子和金葡碱操纵子均与关键传染病状态的发病机制有关，值得进一步研究。
更新日期：2017-11-05
中文翻译：
铜绿假单胞菌对高山金属团的生物合成
细菌的发病机理经常需要通过专门的小分子金属团获得金属。我们假设革兰氏阴性铜绿假单胞菌编码烟碱胺合酶（NAS）和鸦片脱氢酶（ODH）酶，生物合成了一类新型的鸦片金属荧光团，以前仅在无关的革兰氏阳性生物体金黄色葡萄球菌中有特征。通过测量结合亲和力，体外合成筛选潜在底物的生物合成途径以及通过质谱法确认产物形成，来确定该金属团簇的身份，在本文中称为假papalpaline 。伪帕林和金黄色葡萄球菌金属团葡萄球碱对组氨酸部分显示相反的立体化学，表明被NAS唯一识别。此外，我们证明了丙酮酸存在下的SaODH催化作用（如先前所示），草酸乙酸酯也存在，这表明了产生葡萄球菌素变体形式的潜力，而PaODH特异性识别α-酮戊二酸。葡萄球菌操纵子和金葡碱操纵子均与关键传染病状态的发病机制有关，值得进一步研究。
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