Production of selenium nanoparticles occurs through an interconnected pathway of sulphur metabolism and oxidative stress response in Pseudomonas putida KT2440.
Production of selenium nanoparticles occurs through an interconnected pathway of sulphur metabolism and oxidative stress response in Pseudomonas putida KT2440.
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DOI：
10.1111/1751-7915.14215
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发表时间：
2023-05
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
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土壤细菌恶臭假单胞菌KT 2440已被证明从亚硒酸盐有氧产生硒纳米颗粒;然而，参与这一过程的分子演员是未知的。在这里，通过遗传和分析技术的结合，我们报告了这种细菌中亚硒酸盐代谢的第一个见解。我们的研究结果表明，亚硒酸盐的减少发生通过一个相互关联的代谢网络，涉及中央代谢反应，硫代谢和氧化应激反应。sucA、D2 HGDH和PP_3148等基因揭示了2-酮戊二酸和谷氨酸代谢对亚硒酸盐转化为硒很重要。另一方面，影响亚硫酸盐还原酶活性的突变降低了细菌转化亚硒酸盐的能力。其他与硫代谢（ssuEF，sfnCE，sqrR，sqr和pdo 2）和胁迫反应（gqr，lsfA，ahpCF和sadI）相关的基因也被确定为参与亚硒酸盐转化。有趣的是，基因sqrR、sqr和pdo 2的抑制导致硒纳米颗粒的产生速率高于野生型菌株，这具有生物技术意义。这项研究提供的数据使我们更接近于了解硒在细菌中的代谢，并为开发用于生产硒纳米颗粒的生物技术工具提供了新的目标。土壤细菌恶臭假单胞菌KT 2440能够还原亚硒酸盐以形成元素硒的纳米颗粒。在这项工作中，我们结合联合收割机转座筛选与转录组学和生物化学测定，以阐明这一过程的生物技术的兴趣。我们报告说，亚硒酸盐还原依赖于细胞内还原型谷胱甘肽的可用性，并涉及属于硫代谢，中央代谢和氧化应激途径的酶的协同作用。
The soil bacterium Pseudomonas putida KT2440 has been shown to produce selenium nanoparticles aerobically from selenite; however, the molecular actors involved in this process are unknown. Here, through a combination of genetic and analytical techniques, we report the first insights into selenite metabolism in this bacterium. Our results suggest that the reduction of selenite occurs through an interconnected metabolic network involving central metabolic reactions, sulphur metabolism, and the response to oxidative stress. Genes such as sucA, D2HGDH and PP_3148 revealed that the 2‐ketoglutarate and glutamate metabolism is important to convert selenite into selenium. On the other hand, mutations affecting the activity of the sulphite reductase decreased the bacteria's ability to transform selenite. Other genes related to sulphur metabolism (ssuEF, sfnCE, sqrR, sqr and pdo2) and stress response (gqr, lsfA, ahpCF and sadI) were also identified as involved in selenite transformation. Interestingly, suppression of genes sqrR, sqr and pdo2 resulted in the production of selenium nanoparticles at a higher rate than the wild‐type strain, which is of biotechnological interest. The data provided in this study brings us closer to understanding the metabolism of selenium in bacteria and offers new targets for the development of biotechnological tools for the production of selenium nanoparticles. The soil bacterium Pseudomonas putida KT2440 is able to reduce selenite to form nanoparticles of elemental selenium. In this work we combine a transposition screening with transcriptomics and biochemical assays to shed light into this process of biotechnological interest. We report that selenite reduction depends on the intracellular availability of reduced glutathione and involves the concerted action of enzymes belonging to the sulphur metabolism, the central metabolism and oxidative stress pathways.
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