Bioproduction, bioprotection, and biocontainment in multi-kingdom microbial systems with 3D spatial control

Area of Science:

• Biomaterials Engineering
• Synthetic Biology
• Microbial Engineering

Background:

• Engineered living materials (ELMs) combine microbes and polymers, but controlling microbial dynamics in 3D is challenging.
• Current ELMs show biocompatibility, yet spatial organization within 3D form factors requires further investigation.
• Leveraging 3D structures to manage microbial behavior within materials is a key knowledge gap.

Purpose of the Study:

• To explore the use of 3D printing for spatial organization of microbial systems within hydrogel constructs.
• To investigate how 3D form factors influence microbial dynamics, bioproduction, and biocontainment in ELMs.
• To develop customizable multi-kingdom ELMs with precise control over biological functions.

Main Methods:

• Extrusion-based 3D printing was used to fabricate multi-material hydrogel constructs.
• Core-shell cubic geometries were employed to encapsulate single and dual-microbial systems.
• Microbial behavior, including metabolite production, growth, and cell distribution, was monitored over time.

Main Results:

• Successfully fabricated core-shell hydrogel constructs enabling spatial organization of E. coli and S. cerevisiae.
• Demonstrated that 3D spatial organization and surface area-to-volume ratio impact microbial behavior.
• Showcased customizable control over bioproduction, bioprotection, and biocontainment using F127-BUM core-shell geometries.

Conclusions:

• Hierarchical 3D printing of multi-kingdom constructs offers customizable control over ELM functions.
• Optimized core-shell structures can lead to ELMs deployable as compact, sustainable bioreactors.
• This work advances the design and application of 3D-printed engineered living materials.