EFRI E3P: High-throughput synthetic biology approaches for mixed plastic degradation and reutilization

自然界中发现的大量微生物，包括细菌和真菌，种类繁多。这些微生物中的许多可以降解各种聚合材料，如生物质和塑料废物。然而，少数已知的微生物可以缓慢地降解塑料。更有效的塑料降解微生物可以作为工业生物技术来回收废旧塑料材料并创造增值产品。特别是，生物化学回收大容量、难以回收的聚苯乙烯和聚乙烯等塑料的能力，将是克服日益严重的全球环境和健康问题的重要一步。该项目团队最近分离出了几种有希望降解聚苯乙烯和聚乙烯的细菌和真菌菌株。这个多学科团队将共同努力，破译这些微生物是如何降解塑料垃圾的，并利用这些信息来设计细菌细胞，以提高降解效率。他们还将利用不同细菌和真菌的混合物作为一个联合体来提高塑料降解效率，模仿微生物在自然界中如何共同作用来分解复杂的材料。最后，利用新型工程微生物将塑料降解产物转化为高价值化学品。拟议的工作将直接加速微生物驱动的塑料降解和再利用的研究和开发，并将通过微生物生物生产和生物制造解决国家对绿色技术的关键需求。该项目将通过在高中和本科阶段接触多学科科学和工程，为培养下一代研究人员做出贡献。该团队还将创建在线学习模块、演讲和社交网络材料，在学术界和公众之间建立伙伴关系，以交流和提高对塑料废物未来和生物修复潜力的科学意识。该项目的愿景是降解混合废塑料，并利用细菌/真菌联合体和工程细菌联合体将降解产物升级为高价值的化学前体。第一个目标是从富集的聚乙烯和聚苯乙烯降解环境细菌/真菌群落中分离的单个微生物中鉴定关键的塑料降解酶。第二个目标是创造量身定制的合成真菌-细菌联合体，可以生物增强混合聚乙烯/聚苯乙烯的降解。第三个目标是开发能够产生有用的化学前体3-羟基丙酸的工程菌株，并用这些菌株来创建一个塑料降解工程微生物的合成联盟。创新的高通量微流体技术将用于加速发现和测试过程。该项目将为塑料降解的生物化学和塑料降解产品向高价值产品的转化提供新的见解，并显著加快下一代生物修复和绿色生物生产技术的发展。此外，合成生物学策略、稳定的微生物联盟构建策略和高通量微流控平台有望在合成生物学、生物制造和生物技术领域具有广泛的吸引力。预期的结果，集中在聚苯乙烯和聚乙烯混合物上，预计将为现实世界中遇到的其他复杂塑料混合物的生化降解和再利用铺平道路。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The abundance of microorganisms found in nature, including both bacteria and fungi, is extremely diverse. Many of these microorganisms can degrade a variety of polymeric materials such as biomass and plastic waste. However, the few known microorganisms that can degrade plastics do so slowly. More efficient plastic-degrading microorganisms could be leveraged as industrial biotechnology to recycle waste plastic materials and create value-added products. In particular, the ability to biochemically recycle high-volume, difficult-to-recycle plastics like polystyrene and polyethylene would be an important step toward overcoming a mounting global environmental and health concern. The project team has recently isolated several promising bacterial and fungal strains that appear to degrade polystyrene and polyethylene. The multidisciplinary team will work together to decipher how these microorganisms degrade plastic waste and use this information to engineer bacterial cells to improve degradation efficiency. They will also utilize a mixture of different bacteria and fungi as a consortium to improve the plastic degradation efficiency, mimicking how microbes work together in nature to decompose complex materials. Finally, the plastic degradation products will be converted into high-value chemicals using novel engineered microorganisms. The proposed work will directly accelerate the research and development of microbe-driven plastic degradation and re-utilization and will address a critical national need for green technology through microbial bioproduction and biomanufacturing. The project will contribute to the training of next-generation researchers through exposure to multidisciplinary science and engineering at the high school and undergraduate levels. The team will also create online learning modules, presentations, and social network materials that will build partnerships between academia and the public to communicate and enhance the scientific awareness about the future of plastic waste and the potential for bioremediation.The project vision is to degrade mixed waste plastic and upcycle the degradation products into high-value chemical precursors using bacterial/fungal consortia and engineered bacterial consortia. The first aim is to identify the key plastic-degrading enzymes from individual microorganisms isolated from the enriched polyethylene- and polystyrene-degrading environmental bacterial/fungal consortia. The second aim is to create tailored synthetic fungal-bacterial consortia that can bio-augment mixed polyethylene/polystyrene degradation. The third aim is to develop engineered strains that produce a useful chemical precursor, 3-hydroxypropionic acid, and use these to create a synthetic consortium of plastic-degrading engineered microbes. Innovative high-throughput microfluidic technologies will be used to accelerate the discovery and testing processes. The project will yield new insights into the biochemistry of plastic degradation and conversion of plastic degradation products to high-value products, as well as significantly accelerate the development of next-generation bioremediation and green bioproduction technologies. In addition, the synthetic biology strategy, stable microbial consortia construction strategy, and high-throughput microfluidic platforms are expected to have broad appeal in the fields of synthetic biology, biomanufacturing, and biotechnology. The anticipated outcomes, focused here on polystyrene and polyethylene mixtures, are expected to pave the way for biochemical degradation and re-utilization of other complex plastic mixtures encountered in the real world.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.