RT Journal Article
SR Electronic
T1 Metabolism Control in 3D Printed Living Materials
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.01.15.426505
DO 10.1101/2021.01.15.426505
A1 Tobias Butelmann
A1 Hans Priks
A1 Zoel Parent
A1 Trevor G. Johnston
A1 Tarmo Tamm
A1 Alshakim Nelson
A1 Petri-Jaan Lahtvee
A1 Rahul Kumar
YR 2021
UL http://biorxiv.org/content/early/2021/01/17/2021.01.15.426505.abstract
AB The three-dimensional printing of cells offers an attractive opportunity to design and develop innovative biotechnological applications, such as the fabrication of biosensors or modular bioreactors. Living materials (LMs) are cross-linked polymeric hydrogel matrices containing cells, and recently, one of the most deployed LMs consists of F127-bis-urethane methacrylate (F127-BUM). The material properties of F127-BUM allow reproducible 3D printing and stability of LMs in physiological environments. These materials are permissible for small molecules like glucose and ethanol. However, no information is available for oxygen, which is essential— for example, towards the development of aerobic bioprocesses using microbial cell factories. To address this challenge, we investigated the role of oxygen as a terminal electron acceptor in the budding yeast’s respiratory chain and determined its permissibility in LMs. We quantified the ability of cell-retaining LMs to utilize oxygen and compared it with cells in suspension culture. We found that the cells’ ability to consume oxygen was heavily impaired inside LMs, indicating that the metabolism mostly relied on fermentation instead of respiration. To demonstrate an application of these 3D printed LMs, we evaluated a comparative brewing process. The analysis showed a significantly higher (3.7%) ethanol production using 3D printed LMs than traditional brewing, indicating an efficient control of the metabolism. Towards molecular and systems biology studies using LMs, we developed a highly reliable method to isolate cells from LMs for flow cytometry and further purified macromolecules (proteins, RNA, and DNA). Our results show the application of F127-BUM-based LMs for microaerobic processes and envision the development of diverse bioprocesses using versatile LMs in the future.Competing Interest StatementThe authors have declared no competing interest.