Elucidating the mechanism of bacteria-mediated oil degradation by imaging biofilm formation on oil drops in microfluidic traps

通过对微流体捕集器中油滴上的生物膜形成进行成像来阐明细菌介导的油降解机制

• 批准号: 21H01720
• 负责人: UTADA ANDREW
• 金额: $ 11.15万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21H01720/
• 关键词: biofilm; microfluidics; oil degradation; oil-water interface; bioremediation; active matter; living liquid crystal; interface; nematic ordering; biofilm; microfluidics; oil degradation; oil-water interface; bioremediation; active matter; living liquid crystal; interface; nematic ordering

We have investigated cell attachment and early stages of biofilm formation on drops. We have correlated motion between surface-attached cells on the interface. We have found clear liquid crystal ordering of cells on drop surfaces, which indicates that the biofilm is elastic. Further aiding in our interfacial imaging, we have constructed a set of fluorescent and deletion mutant strains. Our constitutive green fluorescent protein (GFP) and mCherry (red fluorescent protein) expressing cells have facilitated imaging of the oil-water interface throughout the process. Moreover, we have measured and analyzed the interfacial properties of the cells and their conditioned components, finding that the cells are more hydrophobic the longer they are cultured using oil and that they develop the ability to enact surface changes.In addition, we have completed RNA-sequencing tests on WT cultured under different conditions and have identified a number of genes that are important for oil consumption. We are currently testing the deletion mutants to see if we can determine the genetic basis of the changes we see.We have also completed a manuscript that describes our results and have uploaded it to the bioRxiv and have submitted it. We are currently undergoing the peer-review process.

我们已经研究了滴在液滴上的生物膜形成的早期阶段。我们在界面上的表面连接细胞之间具有相关的运动。我们已经发现细胞在滴表面上的清晰液晶有序，这表明生物膜是弹性的。在我们的界面成像中进一步协助，我们构建了一组荧光和缺失突变菌株。我们的组成型绿色荧光蛋白（GFP）和表达细胞的MCHERRY（红色荧光蛋白）在整个过程中促进了油水界面的成像。此外，我们已经测量和分析了细胞及其条件成分的界面特性，发现细胞使用油培养的时间越长，并且它们发展了实施表面变化的能力。在另外，我们已经完成了在不同条件下对WT培养的RNA续顺序测试，并且在不同的条件下培养并确定了许多对油的重要性。我们目前正在测试删除突变体，以查看是否可以确定所见变化的遗传基础。我们还完成了一个描述我们结果并将其上传到Biorxiv并提交的手稿。我们目前正在进行同行评审过程。