Dambruin Nicole A, Pronk Jack T, Klijn Marieke E
Department of Biotechnology, Delft University of Technology, Van Der Maasweg 9, Delft, 2629, HZ, The Netherlands.
Anal Bioanal Chem. 2025 Jun 7. doi: 10.1007/s00216-025-05949-2.
Synthetic microbial co-cultures can enhance bioprocess performance by division-of-labor strategies that, through spatial segregation of product-pathway modules, circumvent or mitigate negative impacts of the expression of an entire product pathway in a single microorganism. Relative abundance of the microbial partners is a key parameter for the performance of such co-cultures. Population control strategies based on genetic engineering have been explored, but the required interventions may impose an additional metabolic burden and thereby negatively affect co-culture performance. Regulation of co-culture composition by controlled substrate feeding strategies or temperature control requires real-time population monitoring. Process analytical technology (PAT) is an approach for real-time monitoring and control of processes, enabling continuous observation of co-cultivation that may serve as a foundation for population control strategies. In this review, we discuss PAT methods for monitoring synthetic co-cultures, either through direct biomass measurements or by tracking soluble or volatile metabolites. We discuss advantages, limitations, and applications of established as well as emerging technologies and conclude that leveraging PAT for precise, real-time population control has the potential to enhance stability, efficiency, and industrial scalability of synthetic co-cultures.
合成微生物共培养物可以通过分工策略提高生物过程性能，该策略通过产物途径模块的空间分离，规避或减轻单个微生物中整个产物途径表达的负面影响。微生物伙伴的相对丰度是此类共培养物性能的关键参数。基于基因工程的种群控制策略已被探索，但所需的干预可能会带来额外的代谢负担，从而对共培养性能产生负面影响。通过控制底物进料策略或温度控制来调节共培养物组成需要实时种群监测。过程分析技术（PAT）是一种用于过程实时监测和控制的方法，能够对共培养进行连续观察，这可为种群控制策略奠定基础。在本综述中，我们讨论了通过直接生物量测量或追踪可溶性或挥发性代谢物来监测合成共培养物的PAT方法。我们讨论了现有技术和新兴技术的优点、局限性及应用，并得出结论：利用PAT进行精确的实时种群控制有潜力提高合成共培养物的稳定性、效率和工业可扩展性。
