17-ERACoBioTech - MEmbrane Modulation for BiopRocess enhANcEment - MeMBrane

17-ERACoBioTech - 用于生物过程增强的膜调节 - 膜

• 批准号: BB/R02152X/1
• 负责人: Alan Goddard
• 金额: $ 80.63万
• 依托单位: Aston University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR02152X%2F1
• 关键词: 17; ERACoBioTech; MEmbrane; Modulation; BiopRocess

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.

全球经济对化石原材料的依赖不可持续，工业对原材料投入的需求稳步增长。对环境可持续性的关切正变得越来越严重;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的替代品。使用微生物从可再生生物质中生产材料的细胞工厂是一种有吸引力的替代方案，并且越来越多的平台化学品正在使用工程微生物以工业规模生产。预计这些将对工业生物技术产生变革性影响，但首先，我们必须应对设计和优化高产细胞工厂菌株的挑战，这些菌株可以生产商业上可行的产品。产品产量差的一个原因是生产条件最终对生产细胞有毒。除了对细胞内组分如酶的损害之外，脂质细胞膜和相关蛋白质易受生物分子如乙醇和丙酸盐的影响，以及易受生产过程中的物理参数如渗透压、pH和温度的影响。有一种方法可以使细胞膜在脂质和蛋白质含量方面得到“调整”，使其对毒性带来的压力更具抵抗力，这将彻底改变该领域。此外，表达有效的膜转运蛋白以输出“毒性”产物可以减轻细胞内损伤。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，提高产品产量，降低成本，并有助于推动远离化石原料。采用这种“绿色”工艺和避免消耗不可再生碳源将带来巨大的社会和环境效益。目前由于毒性而在经济上不可行的产品和工艺可以通过菌株抗性的微小增加和伴随的产量增加而变得有利可图。（阿斯顿、约克、FZJ、IATA-CSIC和格罗宁根）和两家大型行业合作伙伴（Lallemand和Pakmaya）在五个国家合作，并在试验规模上验证，设计了强大的细胞工厂（酵母和丙酸杆菌），克服现有的毒性挑战，提高效率，并允许其有效的商业化。该项目中开发的策略将适用于整个行业，以促进具有深远利益的合理应变工程。该项目分为七个相互关联的迭代工作包（WP），采用成熟的构建-测试-分析方法。组学数据的初步分析将确定膜蛋白和脂质含量的关键变化的微生物受到压力与生物生产和那些菌株已知是有点耐这种压力（WP 1）。将使用体外和计算机模拟方法快速描述这些改变的作用，并合理设计更具抗性的膜（WP 2）。使用合成生物学和菌株进化方法，我们将改变微生物的膜组成，以反映WP 2（WP 3）中确定的“最佳”膜。将以高通量方式鉴定最佳菌株，并进行大规模测试，以确保所做的更改转化为工业环境（WP 4）。在此之后，循环的另一次迭代将进一步优化菌株。WP 5将评估创新生产工艺和最终产品的环境和社会可持续性。WP 6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涉及联合体管理、项目治理、沟通活动和行政监督，以确保项目产生最大影响。

项目成果

Detergent-Free Membrane Protein Purification Using SMA Polymer.

• DOI: 10.1007/978-1-0716-2368-8_21
• 发表时间: 2022-01-01
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Broadbent, Luke;Depping, Peer;Rothnie, Alice J
• 通讯作者: Rothnie, Alice J

Membrane Protein Production in the Yeast P. pastoris.

毕赤酵母中的膜蛋白生产。

• DOI: 10.1007/978-1-0716-2368-8_10
• 发表时间: 2022
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Ayub H

Adaptive response to wine selective pressures shapes the genome of a Saccharomyces interspecies hybrid.

• DOI: 10.1099/mgen.0.000628
• 发表时间: 2021-08
• 期刊: Microbial genomics
• 影响因子: 3.9
• 作者: Lairón-Peris M;Castiglioni GL;Routledge SJ;Alonso-Del-Real J;Linney JA;Pitt AR;Melcr J;Goddard AD;Barrio E;Querol A
• 通讯作者: Querol A

Membrane Protein Production in Insect Cells.

昆虫细胞中膜蛋白的产生。

• DOI: 10.1007/978-1-0716-2368-8_12
• 发表时间: 2022
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Vaitsopoulou A

Membrane manipulation by free fatty acids improves microbial plant polyphenol synthesis.

• DOI: 10.1038/s41467-023-40947-x
• 发表时间: 2023-09-12
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Tharmasothirajan, Apilaasha;Melcr, Josef;Linney, John;Gensch, Thomas;Krumbach, Karin;Ernst, Karla Marlen;Brasnett, Christopher;Poggi, Paola;Pitt, Andrew R.;Goddard, Alan D.;Chatgilialoglu, Alexandros;Marrink, Siewert J.;Marienhagen, Jan
• 通讯作者: Marienhagen, Jan