FMSG: Bio: Integrated bioprocess and synthetic biology for future biomanufacturing of industrial products

未来利用新的合成生物学工具和先进的生物工艺将丰富的生物质和废物资源转化为具有可比特性的附加值产品的工业产品的生物制造，将使循环生物经济具有负担得起的能源、经济增长以及可再生能源和化学品生产的创新。在这个项目中，一个多学科团队将合作进行研究，以了解和缓解限制工业发酵连续生产的瓶颈。该项目团队将专注于几种非模型微生物，这些微生物目前被用作生产工业化学品的细胞工厂，或具有巨大的潜力。该项目的结果将指导和加速未来的化学品和燃料的生物制造，并将对美国生物制造行业产生重大和持久的影响。该项目将使农业/农村社区受益，将玉米秸秆等丰富的低价值农业残留物转化为增值产品，并加速可持续生物经济的增长。生物制造还可以减少温室气体(GHG)排放，并对减少气候变化产生重大影响。该项目还将扩大代表性不足群体的参与范围，并培训各种学生和劳动力参与者，使他们具备从事未来生物制造的技能。微生物寿命和老化是影响化学品生产的工业发酵的根本重要现象，但对大多数微生物，包括那些具有重要工业应用的微生物，还没有进行研究。该项目的重点是了解和调节微生物寿命基因和调控途径，在选定的非模型但多功能微生物中利用可再生资源生产化学品和生物燃料。这种方法将结合细胞回收，在连续或半连续(顺序批次/补料批次)发酵中实现高细胞密度和高体积生产率。目前发酵过程中化学品和燃料的生产受到产品效价、生产率或产率(Try)低、过程稳定性差、生产持续时间短(长时间)的限制。这些工艺对于工业应用来说也很昂贵。该项目将研究影响微生物寿命和老化的基因和因素，这些基因和因素影响工业发酵中的细胞活性、工艺性能(Try)和寿命。首先，选择具有工业价值的微生物菌株在不同的培养和胁迫条件下进行评估，通过种群和转录组学分析来研究它们对生长/发酵动力学和培养稳定性/寿命的影响。结果将被用来识别导致培养异质性、生产变异性和有限的生产持续时间或寿命的基因/酶。然后，新的合成生物学工具，包括基于重组酶的状态机(RSM)基因电路和CRISPR基因组工程，将被用于设计菌株，以获得更长的寿命，并通过增强抗逆性来调节衰老。研究假设是，在工业发酵中，寿命延长或衰老减缓的微生物菌株在更长的时间内将更具生产力。这些菌株可以通过设计-构建-测试-学习(DBTL)循环开发，并用于先进的连续发酵过程和原位产品回收，在延长连续生产周期的尝试中实现至少50%的改进。这项未来制造奖得到了生物科学局分子和细胞生物科学部的支持。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。

Future biomanufacturing of industrial products using novel synthetic biology tools and advanced bioprocesses that convert abundant biomass and waste resources into value-added products with comparable properties will enable circular bioeconomy with affordable energy, economic growth, and innovation in renewable energy and chemicals production. In this project, a multidisciplinary team will collaborate on the research to understand and mitigate bottlenecks limiting continuous production in industrial fermentation. The project team will focus on several non-model microorganisms that are currently used or have enormous potential as cell factories for producing industrial chemicals. The results from this project will guide and accelerate future biomanufacturing of chemicals and fuels and would have large and lasting impacts on the US biomanufacturing industry. The project will benefit the agricultural/rural communities by converting abundant low-value agricultural residues such as corn stover to value-added products and accelerate the growth of a sustainable bioeconomy. Biomanufacturing can also reduce greenhouse gas (GHG) emissions and make a major impact on reducing climate change. This project will also broaden the participation of underrepresented groups and train a diverse range of students and workforce participants with skills to engage in future biomanufacturing. Microbial lifespan and aging are fundamentally important phenomena that will impact industrial fermentation for chemicals production but have not been studied for most microbes including those with important industrial applications. This project focuses on understanding and modulating microbial lifespan genes and regulatory pathways in selected non-model but versatile microbes to produce chemicals and biofuels from renewable resources. This approach will integrate cell recycling to achieve high cell density and high volumetric productivity in continuous or semi-continuous (sequential batch/fed-batch) fermentation. Current production of chemicals and fuels in fermentation is limited by low product titer, productivity or rate, and yield (TRY), poor process stability, short production duration (longevity). These processes are also expensive for industrial application. This project will investigate genes and factors affecting microbial lifespan and aging, which impact cell viability, process performance (TRY), and longevity in industrial fermentation. First, selected microbial strains of industrial interest will be evaluated under different culture and stress conditions to study their effects on growth/fermentation kinetics and culture stability/longevity with population and transcriptomics analyses. The results will be used to identify genes/enzymes contributing to culture heterogeneity, production variability, and limited production duration or longevity. Then, novel synthetic biology tools including recombinase-based state machine (RSM) gene circuits and CRISPR genome engineering, will be used to engineer strains for attaining prolonged lifespan and mediated aging via enhanced stress tolerance. The research hypothesis is that microbial strains with increased lifespan or mitigated aging will be more productive for a longer duration in industrial fermentation. Such strains can be developed through the design-build-test-learn (DBTL) cycle and used in advanced continuous fermentation process with in-situ product recovery, achieving at least 50% improvements in TRY for an extended continuous production period.This Future Manufacturing award was supported by the Division of Molecular and Cellular Biosciences in the Directorate for Biological Sciences.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.