The development of a culture medium is an essential step in any bioprocess involving microorganisms for the bioconversion of by‐products to valuable chemicals, making industries like the biofuel industry more competitive. Optimization of the bioconversion process to minimize cost while maximizing yield underscores the importance of using computational methods to identify cellular requirements under specific growth conditions. In this study, a computational approach was proposed as an alternative to optimizing glycerol consumption in one of the most common production chassis, Escherichia coli, specifically strain ATCC 8739. Nineteen compounds were identified as essential for E. coli growth in glycerol. Of these, three reactions associated with nitrogen, phosphorous, and oxygen availability were determined as crucial to reaching high growth and glycerol uptake rates. Based on computational results, a glycerol‐based medium was supplemented with reported common chemical compounds that contain nitrogen or phosphorous (NH₄Cl, Na₂HPO₄, and K₂HPO₄) for further experimental validation. When comparing the supplemented culture medium experimentally with LB medium (Luria Bertani), a two‐fold increment in the glycerol consumption was observed. Transcriptomic analysis of the most promising culture medium reveals that high glycerol utilization under aerobic conditions is dependent on phosphorus to avoid toxicity within the cell because of glycerol‐3‐phosphate generation. The result of this study is a resource to determine nutritional requirements that allow the improvement of the use of raw material for the production of compounds that are attractive to the bio‐based industry. © 2021 Society of Chemical Industry and John Wiley & Sons, Ltd

中文翻译：

---

使用中央碳模型作为替代方法来优化野生型大肠杆菌中的甘油消耗

在涉及微生物的生物过程中，培养基的开发是必不可少的步骤，可将副产物生物转化为有价值的化学物质，从而使生物燃料等行业更具竞争力。优化生物转化过程以最小化成本，同时最大化产量，这突出了使用计算方法来识别特定生长条件下细胞需求的重要性。在这项研究中，提出了一种计算方法来替代最常见的生产底盘大肠杆菌（特别是ATCC 8739菌株）中甘油的最佳消耗量。已鉴定出19种化合物对大肠杆菌是必不可少的甘油的增长。其中，与氮，磷和氧的可利用性有关的三个反应被确定为达到高生长和甘油摄取速率的关键。根据计算结果，以甘油为基础的介质补充了已报告的常见化合物，其中包含氮或磷（NH ₄ Cl，Na ₂ HPO ₄和K ₂ HPO ₄）以进行进一步的实验验证。通过实验将补充培养基与LB培养基（Luria Bertani）进行比较时，观察到甘油消耗量增加了两倍。对最有前途的培养基进行的转录组分析显示，在有氧条件下，甘油的高利用率取决于磷，以避免由于磷酸甘油3-磷酸而在细胞内产生毒性。这项研究的结果为确定营养需求提供了资源，从而可以改善原材料的使用来生产对生物基产业有吸引力的化合物。©2021化学工业协会和John Wiley＆Sons，Ltd