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

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.

我们研究了液滴上的细胞附着和生物膜形成的早期阶段。我们在界面上的表面附着细胞之间进行了相关运动。我们发现液滴表面细胞的液晶排列清晰，这表明生物膜具有弹性。为了进一步帮助我们的界面成像，我们构建了一组荧光和缺失突变株。我们的组成型绿色荧光蛋白 (GFP) 和 mCherry（红色荧光蛋白）表达细胞促进了整个过程中油水界面的成像。此外，我们还测量和分析了细胞及其条件成分的界面特性，发现使用油培养的时间越长，细胞的疏水性就越高，并且它们具有产生表面变化的能力。此外，我们还完成了对不同条件下培养的WT的RNA测序测试，并鉴定了许多对油消耗很重要的基因。我们目前正在测试缺失突变体，看看我们是否能够确定我们所看到的变化的遗传基础。我们还完成了一份描述我们结果的手稿，并将其上传到bioRxiv并提交。我们目前正在进行同行评审过程。

项目成果

Alcanivorax borkumensis Biofilms Enhance Oil Degradation By Interfacial Tubulation

Alcanivorax borkumensis 生物膜通过界面管增强石油降解

• DOI: 10.1101/2022.08.06.503017
• 发表时间: 2022
• 期刊: bioRxiv
• 作者: M. Prasad;N. Obana;S.-Z. Lin;S. Zhao;K. Sakai;C. Blanch-Mercader;J. Prost;N. Nomura;J.-F. Rupprecht*;J. Fattaccioli*;A. S. Utada*
• 通讯作者: A. S. Utada*