Metabolism Control in 3D-Printed Living Materials Improves Fermentation
Metabolism Control in 3D-Printed Living Materials Improves Fermentation
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DOI：
10.1021/acsabm.1c00754
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发表时间：
2021-08-30
影响因子：
4.7
通讯作者：
Kumar, Rahul
中科院分区：
文献类型：
--
作者：
Butelmann, Tobias;Priks, Hans;Kumar, Rahul
含有细胞的聚合物水凝胶的三维（3D）打印创造了生命材料（LMs），为开发生物传感器和生物制造等领域的创新技术提供了一个平台。交联f127 -双氨基甲酸乙酯（F127-BUM）的高分子材料特性允许可重复的3D打印和生理条件下的稳定性，使其适合制造lm。尽管基于f127 - bum的LMs允许葡萄糖和乙醇等溶质分子扩散，但仍不清楚这些溶质分子是否允许用于呼吸所必需的氧气。为了确定氧容许度，我们量化了芽殖酵母负载f127 - bum的LMs的溶解氧消耗。此外，我们获得了保留细胞的LMs的数据，这使得LMs和悬浮培养之间可以进行直接比较。我们进一步开发了一种高度可靠的方法从lm中分离细胞，用于流式细胞术分析，细胞活力评估和大分子纯化。我们发现LMs内的氧气消耗严重受损，这表明酵母的代谢主要依赖于发酵而不是呼吸。将这一发现应用于酿造，我们观察到使用LMs比传统酿造工艺更高（3.7%）的乙醇产量，表明发酵得到了改善。我们的研究表明，目前基于f127 - bum的LMs在微好氧过程中是有用的，但开发好氧生物过程需要进一步的研究。
The three-dimensional (3D) printing of cell-containing polymeric hydrogels creates living materials (LMs), offering a platform for developing innovative technologies in areas like biosensors and biomanufacturing. The polymer material properties of cross-linkable F127-bis-urethane methacrylate (F127-BUM) allow reproducible 3D printing and stability in physiological conditions, making it suitable for fabricating LMs. Though F127-BUM-based LMs permit diffusion of solute molecules like glucose and ethanol, it remains unknown whether these are permissible for oxygen, essential for respiration. To determine oxygen permissibility, we quantified dissolved oxygen consumption by the budding yeast-laden F127-BUM-based LMs. Moreover, we obtained data on cell-retaining LMs, which allowed a direct comparison between LMs and suspension cultures. We further developed a highly reliable method to isolate cells from LMs for flow cytometry analysis, cell viability evaluation, and the purification of macromolecules. We found oxygen consumption heavily impaired inside LMs, indicating that yeast metabolism relies primarily on fermentation instead of respiration. Applying this finding to brewing, we observed a higher (3.7%) ethanol production using LMs than the traditional brewing process, indicating improved fermentation. Our study concludes that the present F127-BUM-based LMs are useful for microaerobic processes but developing aerobic bioprocesses will require further research.