Loiko Nataliya, Tereshkina Ksenia, Kovalenko Vladislav, Moiseenko Andrey, Tereshkin Eduard, Sokolova Olga S, Krupyanskii Yurii
Winogradsky Institute of Microbiology, Fundamentals of Biotechnology Federal Research Center, Russian Academy of Sciences, 117312 Moscow, Russia.
Semenov Federal Research Center for Chemical Physics, Russian Academy of Sciences, 119991 Moscow, Russia.
Biology (Basel). 2023 Jun 14;12(6):853. doi: 10.3390/biology12060853.
Gradual dehydration is one of the frequent lethal yet poorly understood stresses that bacterial cells constantly face in the environment when their micro ecotopes dry out, as well as in industrial processes. Bacteria successfully survive extreme desiccation through complex rearrangements at the structural, physiological, and molecular levels, in which proteins are involved. The DNA-binding protein Dps has previously been shown to protect bacterial cells from many adverse effects. In our work, using engineered genetic models of to produce bacterial cells with overproduction of Dps protein, the protective function of Dps protein under multiple desiccation stresses was demonstrated for the first time. It was shown that the titer of viable cells after rehydration in the experimental variants with Dps protein overexpression was 1.5-8.5 times higher. Scanning electron microscopy was used to show a change in cell morphology upon rehydration. It was also proved that immobilization in the extracellular matrix, which is greater when the Dps protein is overexpressed, helps the cells survive. Transmission electron microscopy revealed disruption of the crystal structure of DNA-Dps crystals in cells that underwent desiccation stress and subsequent watering. Coarse-grained molecular dynamics simulations showed the protective function of Dps in DNA-Dps co-crystals during desiccation. The data obtained are important for improving biotechnological processes in which bacterial cells undergo desiccation.
渐进性脱水是细菌细胞在其微生态位干涸时在环境中以及在工业过程中经常面临的致命但了解甚少的应激之一。细菌通过结构、生理和分子水平的复杂重排成功地在极端干燥条件下存活,其中蛋白质发挥了作用。此前已证明DNA结合蛋白Dps可保护细菌细胞免受多种不利影响。在我们的工作中,使用工程遗传模型来生产过量表达Dps蛋白的细菌细胞,首次证明了Dps蛋白在多种干燥应激下的保护功能。结果表明,在Dps蛋白过表达的实验变体中,复水后活细胞的滴度高出1.5至8.5倍。使用扫描电子显微镜显示复水后细胞形态的变化。还证明了在细胞外基质中的固定化(当Dps蛋白过表达时固定化程度更高)有助于细胞存活。透射电子显微镜显示经历干燥应激和随后浇水的细胞中DNA-Dps晶体的晶体结构被破坏。粗粒度分子动力学模拟显示了干燥过程中Dps在DNA-Dps共晶体中的保护功能。所获得的数据对于改进细菌细胞经历干燥的生物技术过程很重要。