Enzyme induced polymer degradation as a pathway to create microbial habitats for biomanufacturing intensification

• Published in: Chemical engineering journal (Lausanne, Switzerland : 1996) Vol. 495; p. 153622
• Main Authors: Cheng, Wei, Wen, Huilin, Shen, Haixia, Fu, Chenwei, Miao, Erkuang, Li, Dongbao, Chen, Xiaoqiang, Hu, Chi, Yu, Ziyi
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.09.2024
• ISSN: 1385-8947
• DOI: 10.1016/j.cej.2024.153622

We introduce an innovative liquid microcavity technique to create microbial habitats. Our successful application of this technology in fabricating yeast-embedded 3D lattices for ethanol production demonstrates its potential to revolutionize biomanufacturing, opening new pathways for the development of advanced biosensors, bioreactors, and other bioactive systems. [Display omitted] •Liquid microcavity was generated in situ by enzyme-catalyzed degradation of gelatin.•F127 hydrogel with liquid cavity serves as high-fidelity bioink without microbial leakage.•Yeast-embedded 3D lattices are fabricated for biocatalytic production of ethanol. Polymer hydrogels are instrumental in cell immobilization for biomanufacturing, yet traditional hydrogel bioreactors often compromise cell viability, function, and mass transfer. This results in uncontrolled microbial proliferation and significant cell leakage, thereby impeding the continuous production of high-value products. To address these challenges, we introduce an innovative liquid microcavity technique employing a blend of Gelatin-methacryloyl (Gel-MA) and Pluronic F127diacrylate (F127-DA) for precision bioprinting, followed by selective enzymatic digestion of gelatin to create microbial habitats. Through tailoring of the hydrogel’s pore structure and integration of diverse networks, our approach facilitates controlled nutrient transfer, efficient waste removal, and stable mechanical support, all while maintaining a leak-free suspension culture of microorganisms. We validated the effectiveness of our method by fabricating yeast-infused Three-dimensional (3D) lattice constructs, successfully achieving continuous fermentation of glucose into ethanol in a flow reactor with minimal efficiency reduction (≥90 % yield) over five cycles. This research not only confronts the spatial limitations inherent in hydrogel-based systems but also demonstrates a foundational potential for revolutionary change in biomanufacturing practices.