Identification of novel catabolic genes involved in 17β-estradiol degradation by Novosphingobium sp. ES2-1
Identification of novel catabolic genes involved in 17β-estradiol degradation by Novosphingobium sp. ES2-1
复制标题
鉴定参与Novosphingobium sp. 17 β-雌二醇降解的新型分解代谢基因。
DOI：
10.1111/1462-2920.15475
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发表时间：
2021-03-26
影响因子：
5.1
通讯作者：
Ling, Wanting
中科院分区：
文献类型：
--
作者：
Li, Shunyao;Sun, Kai;Ling, Wanting
新鞘氨醇杆菌属（Novosphingobium）ES2 - 1是一种高效降解17β - 雌二醇（E2）的细菌，它能将E2转化为雌酮（E1），进而转化为4 - 羟基雌酮（4 - OH - E1）以便后续的氧化裂解。在本研究中，这一过程的分子基础得以阐明。两种新型单加氧酶系统EstP和EstO分别被证明可催化E1和4 - OH - E1的加氧反应。EstP是一个由EstP1（细胞色素P450单加氧酶）、EstP2（铁氧化还原蛋白）和EstP3（铁氧化还原蛋白还原酶）组成的三组分细胞色素P450单加氧酶系统。超高效液相色谱 - 高分辨率质谱（UPLC - HRMS）分析表明，EstP催化E1的4 - 羟基化生成4 - OH - E1。生成的4 - OH - E1进一步被一个由EstO1（黄素依赖型单加氧酶）和EstO2（黄素还原酶）组成的双组分单加氧酶系统EstO氧化。UPLC - HRMS结合氢 - 1 - 核磁共振分析证明，EstO催化C9 - C10键断裂，产生一个B环裂解产物。此外，当暴露于不同的类固醇时，加氧酶组分基因estP1和estO1表现出相反的诱导行为，这表明EstP1介导的4 - 羟基化是E2特异性的，而EstO1介导的单加氧反应可能参与睾酮、雄烯二酮、孕酮和孕烯醇酮的降解。这也意味着同一微生物对不同类固醇的分解代谢机制可能存在部分关联。
Novosphingobium sp. ES2-1 is an efficient 17 beta-estradiol (E2)-degrading bacterium, which can convert E2 to estrone (E1), then to 4-hydroxyestrone (4-OH-E1) for subsequent oxidative cracking. In this study, the molecular bases for this process were elucidated. Two novel monooxygenase systems EstP and EstO were shown to catalyse the oxygenation of E1 and 4-OH-E1, respectively. EstP was a three-component cytochrome P450 monooxygenase system consisting of EstP1 (P450 monooxygenase), EstP2 (ferredoxin) and EstP3 (ferredoxin reductase). Ultraperformance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) analysis revealed that EstP catalysed the 4-hydroxylation of E1 to produce 4-OH-E1. The resultant 4-OH-E1 was further oxidized by a two-component monooxygenase system EstO consisting of EstO1 (flavin-dependent monooxygenases) and EstO2 (flavin reductase). UPLC-HRMS combined with H-1-nuclear magnetic resonance analysis demonstrated that EstO catalysed the breakage of C9-C10 to yield a ring B-cleavage product. In addition, the oxygenase component genes estP1 and estO1 exhibited contrary inductive behaviours when exposed to different steroids, suggesting that EstP1-mediated 4-hydroxylation was E2-specific, whereas EstO1-mediated monooxygenation might be involved in the degradation of testosterone, androstenedione, progesterone and pregnenolone. This also implied that the mechanisms of the catabolism of different steroids by the same microorganism might be partially interlinked.