17-ERACoBioTech - MEmbrane Modulation for BiopRocess enhANcEment - MeMBrane

全球经济对化石原材料的依赖是不可持续的，工业对原材料投入的需求稳步增长。对环境可持续性的担忧正变得更加尖锐；因此，迫切需要替代传统的、以化石燃料为基础的化学品生产。使用微生物从可再生生物质中生产材料的细胞工厂是一个有吸引力的替代方案，越来越多的平台化学品正在使用工程微生物进行工业规模的生产。这些预计将对工业生物技术产生变革性的影响，但首先，我们必须应对设计和优化能够生产商业上可行的产品的高产细胞工厂菌株的挑战。产品产量不佳的一个原因是，生产条件最终会对生产细胞产生毒性。除了对细胞内成分如酶的破坏外，脂质细胞膜和相关蛋白还容易受到生物分子如乙醇和丙酸的影响，以及生产过程中的物理参数如渗透胁迫、pH和温度的影响。一种可以根据膜的脂肪和蛋白质含量对膜进行“调谐”，以提高对毒性带来的压力的抵抗力的方法，将使该领域发生革命性的变化。此外，表达有效的膜转运蛋白以出口“有毒”产品可以减轻细胞内的损害。这些方法最终将能够生产更高浓度的所需分子或细胞，使生物过程更有效，增加产品产量，降低成本，并有助于推动摆脱以化石为基础的原材料。采用这种“绿色”程序，避免耗尽不可再生的碳源，将带来巨大的社会效益和环境效益。在这个为期36个月的项目中，五个领先的研究机构(阿斯顿、约克、FZJ、IATA-CSIC和Groningen)以及五个国家的两个大型行业合作伙伴(拉勒曼德和帕克马亚)将在中试规模上合作并验证健壮的工程细胞工厂(酵母和丙酸杆菌)，以克服现有的毒性挑战，提高效率，并使其有效商业化。该项目制定的战略将适用于整个部门，以促进合理应变工程，带来深远的好处。该项目分为七个相互关联的迭代工作包(WPS)，具有完善的构建-测试-分析方法。对组学数据的初步分析将确定受到与生物生产相关的压力的微生物和已知对这种压力有一定抵抗力的菌株的膜蛋白和膜脂含量的关键变化(WP1)。在体外和在硅胶中的方法将被用来快速描述这些改变的作用，并合理地设计更耐药的膜(WP2)。利用合成生物学和菌株进化的方法，我们将改变微生物的膜组成，以反映WP2(WP3)中确定的“最佳”膜。将以高通量方式确定最佳菌株，并进行大规模测试，以确保所做的改变转化为工业环境(WP4)。之后，循环的另一次迭代将进一步优化菌株。WP5将评估创新生产过程和最终产品的环境和社会可持续性。工作组将制定和实施一项向不同利益攸关方传播和利用研究成果的战略。WP7涉及财团管理、项目治理、沟通活动和行政监督，以确保项目产生最大影响。

The global economy has an unsustainable dependence on fossil raw material with demand for raw material inputs to industry growing steadily. Concerns about environmental sustainability are becoming more acute; thus, alternatives to traditional, fossil-fuel based chemical production are urgently required. Cell factories, which use microorganisms to produce materials from renewable biomass, are an attractive alternative, and an increasing number of platform chemicals are being produced at industrial scale using engineered microorganisms. These are expected to have a transformative impact in industrial biotechnology, but, first, we must meet the challenges of designing and optimizing high-yield cell factory strains that can produce commercially viable amounts of product. One reason for poor product output is that the production conditions are ultimately toxic to the producing cells. In addition to damage to intracellular components such as enzymes, the lipid cell membrane and associated proteins are vulnerable to biomolecules e.g. ethanol and propionate, as well as to physical parameters during production such as osmotic stress, pH, and temperature. An approach whereby membranes can be "tuned", in terms of their lipid and protein content, to become more resistant to stresses brought about by toxicity would revolutionise the field. Additionally, expression of efficient membrane transporters to export 'toxic' products can mitigate intracellular damage. These approaches will ultimately enable production of higher concentrations of the desired molecules or cells making the bioprocesses more efficient, increasing product yield, reducing cost, and help to drive the move away from fossil-based raw materials. An adoption of such "green" processes and avoidance of depletion of non-renewable carbon sources will bring huge social and environmental benefits. Products and processes which are currently economically unviable due to toxicity can be rendered profitable by even small increases in the resistance of strains and concomitant yield increases.This 36-month project sees five leading research institutes (Aston, York, FZJ, IATA-CSIC and Groningen) and two large industry partners (Lallemand and Pakmaya), across five countries, collaborate, and validate at pilot scale, engineered robust cell factories (yeast and Propionibacterium) that overcome existing toxicity challenges, improve efficiency and allow their effective commercialisation. The strategies developed within this project will be applicable across the sector to facilitate rational strain engineering with far-reaching benefits.The project is divided into seven interconnected, iterative work packages (WPs) with a well-established build-test-analyse approach. Initial analysis of -omics data will identify key alterations in membrane protein and lipid content of both microbes subjected to stresses associated with bioproduction and those strains known to be somewhat resistant to such stresses (WP1). In vitro and in silico approaches will be used to rapidly delineate the roles of these alterations and rationally design more resistant membranes (WP2). Using synthetic biology and strain evolution approaches, we will alter the membrane composition of microbes to reflect the "optimal" membranes determined in WP2 (WP3). Optimal strains will be identified in a high throughput manner and subjected to large-scale testing to ensure that the changes made translate to the industrial setting (WP4). Following this, another iteration of the cycle will further optimise the strains. WP5 will evaluate the environmental and social sustainability of the innovative production processes and the final products. WP6 will develop and implement a strategy for the dissemination and exploitation of research results to different stakeholders. WP7 involves consortium management, project governance, communication activities and administrative oversight to ensure maximum impact of the project.