Excellence in Research: Oxidative stress induced impact of cell-penetrating nanoparticles on cellular constituents in a cyanobacterial model

化石燃料的使用造成了不利的环境危害，引发了人们对替代能源技术的浓厚兴趣。作为光合作用微生物，蓝藻可以产生可转化为高能生物燃料的脂类，因此在工业和环境应用方面具有巨大的潜力。这项具有变革性的研究试图了解纳米技术方法在加速开发生物质衍生燃料生产技术以促进环境可持续性方面具有前所未有的能力。铁纳米颗粒由于其高度的化学反应活性而吸附在蓝藻细胞表面，并可产生氧化应激导致代谢变化。这项研究的目标是了解生物-纳米颗粒相互作用在细胞水平上的独特影响及其对工业有用分子生产的影响。本科生和研究生将在母校(摩根州立大学)和国家高磁场实验室(佛罗里达州立大学)接受尖端跨学科研究的指导和培训。该项目产生的研究方案将被纳入调查人员教授的400级本科实验室教育课程，为没有机会进行个性化研究的学生提供丰富的研究经验。通过该项目产生的发现将通过在国家会议、专利、同行评议出版物和外联活动上的陈述来传播。全面了解铁纳米颗粒在蓝藻模型中的作用将对推进研究和促进跨多个学科的发现具有深远的好处，包括生物能源、环境安全补救和生物传感。这项研究的重点是蓝藻中纳米颗粒介导的影响领域中一个尚未解决的主要问题，特别是因为这些生物被用作生物能源的平台。该项目旨在了解零价铁纳米颗粒如何惯性地渗透到蓝藻细胞中，诱导氧化应激，并影响光合色素沉积、蛋白质调节和脂类分布。它将作为未来贡献的基础，通过(I)评估纳米颗粒诱导的应激对蓝藻Fremyella diplosiphon中的活性氧物种和色素积累的影响，(Ii)使用大数据分析破译纳米处理的Fremyella diplosiphon中抗氧化酶的差异蛋白质调控，以及(Iii)使用全面的二维气相色谱-飞行时间和傅立叶变换离子回旋共振质谱仪解开独特的脂肪酸甲酯图谱和极性脂类。首席研究员的研究小组提出了一种混合系统，通过加入铁纳米颗粒来增强蓝藻细胞中的脂质生产，从而产生一种环境安全的替代能源。通过这种创新的方法，该项目将提供一个明确的理解，当纳米铁非对抗性进入细胞时，纳米铁诱导的氧化应激对蛋白质和脂类结构域的影响。解开纳米颗粒在代谢过程中诱导应激反应的机制，将为理解它们在改变细胞成分中的作用提供巨大的潜力。该团队将为学生提供有关项目的互动机会，以生成数据、讨论结果、撰写手稿并在科学会议上展示发现。这个集生物学、工程学、纳米技术和化学为一体的多学科项目将为下一代研究人员提供装备，以应对STEM领域的挑战。建议的策略将产生铁纳米颗粒介导的细胞反应的基础知识，并通过研究氧化应激对细胞过程的影响提供新的见解。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Adverse environmental hazards caused by fossil fuel use have sparked significant interest in alternative energy technologies. As photosynthetic microorganisms, cyanobacteria produce lipids, which can be converted to high-energy biofuels, thus offering great potential for industrial and environmental applications. This transformative research seeks to understand the unprecedented ability of nanotechnological approaches to accelerate the development of biomass-derived fuel production technologies for environmental sustainability. Iron nanoparticles adsorb to cyanobacterial cell surfaces due to their high chemical reactivity and can create oxidative stress leading to metabolic changes. The goal of this study is to understand the unique effects of biological-nanoparticle interactions at the cellular level and their impact on the production of industrially useful molecules. Undergraduate and graduate students will be mentored and trained on cutting-edge interdisciplinary research at the home institution (Morgan State University) and the National High Magnetic Field Laboratory (Florida State University). Research protocols generated from the project will be incorporated in a 400-level undergraduate laboratory-based educational course that the investigator teaches, providing rich research experience for students who do not have the opportunity for individualized research. Discoveries generated through this project will be disseminated through presentations at national conferences, patents, peer-reviewed publications, and outreach activities. A comprehensive understanding of the role of iron nanoparticles in a model cyanobacterium will have far-reaching benefits to advance research and promote discoveries across multiple disciplines including bioenergy, environmentally safe remediation, and biosensing. The research focuses on a major unsolved problem in the field of nanoparticle-mediated impact in cyanobacteria, especially since these organisms are used as a platform for bioenergy. The project aims to understand how zero-valent iron nanoparticles which inertly penetrate cyanobacterial cells can induce oxidative stress, and impact photosynthetic pigmentation, protein regulation, and lipid profile. It will serve as a foundation for future contributions by (i) evaluating the impact of nanoparticle-induced stress on reactive oxygen species and pigment accumulation in cyanobacterium Fremyella diplosiphon, (ii) deciphering differential protein regulation of antioxidative enzymes in nano-treated F. diplosiphon using big data analytics, and (iii) unraveling unique fatty acid methyl ester profiles and polar lipids using comprehensive two-dimensional gas chromatography-time of flight and Fourier transform ion cyclotron resonance mass spectrometry. The principal investigator's research group proposes a hybrid system by incorporating iron nanoparticles to enhance lipid production in cyanobacterial cells, leading to an environmentally-safe alternative energy source. Through this innovative approach, the project will provide a clear understanding of the impact of iron nanoparticle-induced oxidative stress on protein and lipid domains when iron nanoparticles non-antagonistically enter the cell. Unlocking the mechanisms of nanoparticle-induced stress response on metabolic processes will offer tremendous potential to understand their role in altering cellular constituents. The team will provide interactive opportunities to students on the projects to generate data, discuss results, write manuscripts, and present findings at scientific conferences. This multidisciplinary project at the nexus of Biology, Engineering, Nanotechnology, and Chemistry will equip the next-generation researchers to address challenges in STEM fields. The proposed strategies will generate fundamental knowledge of iron nanoparticle-mediated cell response and provide new insights by studying oxidative stress-mediated effects on cellular processes.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.