Engineering Rhodopseudomonas palustris for enhanced biohydrogen production

改造沼泽红假单胞菌以提高生物氢产量

• 批准号: 2763729
• 金额: --
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2763729
• 关键词: Engineering; Rhodopseudomonas; palustris; enhanced; biohydrogen

Fossil fuels (oil, coal, and natural gas) dominate global energy sources but emit CO2, contributing to climate change. Their extraction harms the environment and is finite. Transitioning to cleaner, sustainable energy is essential. Renewables like wind, solar, hydro, and geothermal, plus alternative fuels (e.g., hydrogen and biofuels), are gaining traction. Hydrogen is promising. It's carbon-neutral and versatile, aiding decarbonization. Current hydrogen production, though, relies on fossil fuels, emitting CO2. Clean, renewable options like biohydrogen, produced biologically from organic matter using microbes, offer lower carbon footprints, energy efficiency, and waste management solutions.Microbial electrochemical technologies, notably microbial electrochemical cells (MECs), are used for biohydrogen production. Challenges include enhancing yield and production rate. Addressing these hinges on understanding microbial communities involved in biohydrogen production.Rhodopseudomonas palustris or R. palustris is one of the most attractive and potential candidates commonly utilised in MECs for biohydrogen production. It can fix both carbon and nitrogen. Hydrogen production is the side product of nitrogen fixation. The knowledge gap lies in how the metabolic modules of nitrogen fixation and hydrogen production are controlled in this bacterium. In this context, my project focuses on understanding the interaction among the genes responsible for hydrogen production in R. palustris and engineer them for enhanced biohydrogen production. Work package 1 Task 1.1- Characterisation of different industrial wastewater streams Wastewater from such industries will be explored and characterised for organic content. This will help to identify the wastewater streams that are rich in organic content. Task 1.2- Assessment of different strains of R. palustris that have shown hydrogen productionPreviously studied strains of R. palustris will be inoculated in the different waste streams and the extent of hydrogen production will be documented, enabling us to identify the best hydrogen producing strain. Work package 2Task 2.1- Identification of differentially expressed/regulated pathways The best hydrogen producing strain will be grown under nitrogen limiting or non-limiting conditions and will be subjected to comparative proteomic and transcriptomic analysis.Task 2.2- Validation of genes/regulatory proteins involved in hydrogen production Newly identified pathway components will be subjected to gene inactivation and later functional complementation to elucidate their role in H2 production. Work package 3 Task 3.1- Biological engineering Once the regulation of differentially expressed genes in the best characterised strain of R. palustris responsible for H2 production has been identified, the potential candidate genes will be subjected to further genetic modification (plasmid- based overexpression, genome integration, deregulation, deletion) with the aim of enhancing H2 production. Work package 4- Assessing hydrogen production in engineered R. palustris' strain in MEC. Task 4.1- MEC construction For this task, a two-chambered MEC system separated by a PEM (Nafion (Du Pont) will be used. Task 4.2- Evaluation of genetically engineered R. palustris' strain in MEC for improved hydrogen production The genetically modified strain vs its wildtype will be analysed for hydrogen production in the MEC constructed above. Work package 5 Task 5.1- With the aim to achieve improved hydrogen production, the MEC reactor will be optimised on the following factors:pH, Temperature, Electrode material. Work package 6 Task 6.1 Performing LCA To assess the environmental impact of MEC-based microbial hydrogen production compared to traditional methods, an LCA, from raw material utilisation to disposal, can be executed

化石燃料（石油、煤炭和天然气）在全球能源中占主导地位，但会排放二氧化碳，导致气候变化。它们的开采会危害环境并且是有限的。转向更清洁、可持续的能源至关重要。风能、太阳能、水力和地热能等可再生能源以及替代燃料（例如氢和生物燃料）正在受到越来越多的关注。氢很有前途。它是碳中和且用途广泛，有助于脱碳。然而，目前的氢气生产依赖化石燃料，排放二氧化碳。清洁、可再生的选择，如利用微生物从有机物中生物生产的生物氢，可提供较低的碳足迹、能源效率和废物管理解决方案。微生物电化学技术，特别是微生物电化学电池 (MEC)，用于生物氢生产。挑战包括提高产量和生产率。解决这些问题取决于了解参与生物氢生产的微生物群落。沼泽红假单胞菌或沼泽红假单胞菌是 MEC 中常用于生物氢生产的最具吸引力和潜力的候选者之一。它可以固定碳和氮。氢气生产是固氮的副产品。知识差距在于如何控制这种细菌的固氮和产氢代谢模块。在这种背景下，我的项目重点是了解沼泽红藻中负责产氢的基因之间的相互作用，并对它们进行改造以增强生物氢的产量。工作包 1 任务 1.1- 不同工业废水流的表征 将探索这些工业废水的有机含量并对其进行表征。这将有助于识别富含有机物的废水流。任务 1.2- 评估已显示产氢的不同沼泽红藻菌株 先前研究的沼泽红藻菌株将接种在不同的废物流中，并且将记录产氢程度，使我们能够确定最佳产氢菌株。工作包2 任务2.1-差异表达/调节途径的鉴定 最好的产氢菌株将在氮限制或非限制条件下生长，并将进行比较蛋白质组和转录组分析。任务2.2-参与产氢的基因/调节蛋白的验证 新鉴定的途径组分将进行基因失活和随后的功能互补，以阐明其作用 在氢气生产中。工作包 3 任务 3.1-生物工程 一旦确定了负责 H2 生产的沼泽沼泽菌中差异表达基因的调控，潜在的候选基因将受到进一步的遗传修饰（基于质粒的过表达、基因组整合、解除管制、删除），以提高 H2 产量。工作包 4-评估 MEC 中工程 R. palustris 菌株的氢气生产。任务 4.1-MEC 构建 对于此任务，将使用由 PEM（Nafion（杜邦）分隔的两室 MEC 系统。任务 4.2-评估 MEC 中的基因工程 R. palustris 菌株以提高产氢量。将分析转基因菌株与其野生型在上面构建的 MEC 中的产氢量。工作包 5 任务 5.1-为了实现提高产氢量， MEC反应器将根据以下因素进行优化：pH、温度、电极材料。工作包 6 任务 6.1 执行 LCA 为了评估基于 MEC 的微生物制氢与传统方法相比对环境的影响，可以执行从原材料利用到处置的 LCA