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

未来工业产品的生物制造使用新型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增值产品，将使循环生物经济成为可能，并带来负担得起的能源，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研究，以了解和减轻限制工业发酵连续生产的瓶颈。该项目团队将专注于目前使用或具有巨大潜力的几种非模型微生物，作为生产工业化学品的细胞工厂。该项目的结果将指导和加速未来化学品和燃料的生物制造，并将对美国生物制造业产生巨大而持久的影响。该项目将使农业/农村社区受益，将大量的低价值农业残留物（如玉米秸秆）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的群体的参与，并培训各种学生和劳动力参与者，使其具备参与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基本重要现象，但尚未对大多数微生物进行研究，包括具有重要工业应用的微生物。该项目的重点是了解和调节选定的非模型但多才多艺的微生物中的微生物寿命基因和调控途径，以从可再生资源中生产化学品和生物燃料。该方法将整合细胞再循环以在连续或半连续（连续分批/补料分批）发酵中实现高细胞密度和高体积生产率。目前发酵中化学品和燃料的生产受到低产品滴度、生产率或速率和产率（TRY）、差的工艺稳定性、短生产持续时间（寿命）的限制。这些方法对于工业应用来说也是昂贵的。该项目将研究影响微生物寿命和老化的基因和因素，这些因素会影响细胞活力，工艺性能（TRY）和工业发酵中的寿命。首先，将在不同的培养和强制降解条件下评价工业感兴趣的选定微生物菌株，以研究其对生长/发酵动力学和培养物稳定性/寿命的影响，并进行群体和转录组学分析。结果将用于鉴定导致培养物异质性、生产变异性和有限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，将用于工程菌株，以通过增强应激耐受性来延长寿命和介导衰老。研究假设是，寿命延长或老化减缓的微生物菌株在工业发酵中的生产力将更高，持续时间更长。这样的菌株可以通过设计-构建-测试-学习（DBTL）循环来开发，并用于具有原位产物回收的先进连续发酵工艺中，达到至少50%改进TRY以延长连续生产期。该未来制造奖得到了生物科学理事会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，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，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.