bims-livmat 2026-10-25 papers

ACS Appl Bio Mater. 2026 Oct 19.

Additive Manufacturing of Metabolically Active Living Materials with Enhanced Biocontainment.

Henrique Sepúlveda Del Rio Hamacek, Shannon Daily, LeAnn Le, Sadia Khalid, Pirjo Spuul, Petri-Jaan Lahtvee, Alshakim Nelson, Rahul Kumar.

Additive manufacturing of engineered living materials (ELMs) enables the spatial patterning of living cells or organisms within three-dimensional (3D) polymer matrices. However, effective biocontainment, defined as the ability to retain cells within 3D-printed matrices during the application period, remains a key limitation for ELMs requiring physical confinement. While synthetic biology provides cell-specific biocontainment strategies, materials-based approaches can enable generic cell retention capabilities. Therefore, we applied a materials-based strategy that combines pre-printing formulation development and post-printing processing to increase the retention of yeast Saccharomyces cerevisiae in Pluronic F127-bisurethane methacrylate (F127-BUM) hydrogels. F127-BUM precursor solutions were formulated with the comonomers methacrylic acid (MAA) and/or 2-(dimethylamino)ethyl methacrylate (DMAEMA), extrusion-printed, UV-cross-linked, and immersed in 70 vol % ethanol for 60 or 300 s before cultivation. Comonomer addition preserved dimensional fidelity, thermoresponsiveness, and shear-thinning behavior for 3D printing. After immobilization and cross-linking, DMAEMA and higher concentrations of MAA considerably reduced yeast viability, whereas a 70-80% viability was observed for 0.03 and 0.06 mol L-1 MAA. With 0.06 mol L-1 MAA and 300 s ethanol immersion, the yeast retention time reached 98 ± 12 h, which was approximately 48% longer than that of the correspondingly treated F127-BUM-only control and represented about a 465% increase relative to the untreated F127-BUM-only control. After ethanol immersion, a peripheral growth-free zone developed around a metabolically active core, enhancing cell retention and contributing to delayed cell escape. The selected MAA-containing ELMs remained metabolically active and showed nearly 38% higher apparent glucose-to-ethanol conversion than treated F127-BUM-only controls. The suspension cultures fermented faster and achieved higher yield. Our method achieved enhanced biocontainment, with an operational window of about 4-5 days. In conclusion, we demonstrate that combining pre-printing and post-printing processes can enhance biocontainment for different ELM applications. We suggest the integration of this method with synthetic-biology strategies to further extend biocontainment duration for metabolically active living materials.

Keywords: 3D-printed bioreactors, metabolically active living materials; Pluronic F127; additive manufacturing; biocontainment; bioproduction; cell retention; engineered living materials; fermentation; hydrogels

DOI: https://doi.org/10.1021/acsabm.6c01394

Proc Natl Acad Sci U S A. 2026 Oct 27. 123(43): e2624792123

Signal peptides for engineered protein secretion by free and hydrogel-encapsulated cyanobacteria.

Nathan T Soulier, Debika Datta, Rakefet Schwarz, Jonathan K Pokorski, James W Golden, Susan S Golden.

Cyanobacteria provide a promising platform for light-driven heterologous protein production. In heterologous production systems, cells are typically lysed for product purification or to facilitate interaction with external substrates, unless the product is secreted. Fusion to molecular "shipping labels" known as signal peptides can target chimeric proteins to native membrane translocation pathways and result in secretion. In this work, we identified signal peptides from the model cyanobacterium Synechococcus elongatus PCC 7942 (S. elongatus) and genetically fused to two heterologous enzymes: the reporter nanoluciferase (NLuc) and an oxidative laccase (CotA). Increased extracellular activity was observed for one or more signal-protein variants for both enzymes. Furthermore, cells of S. elongatus producing a signal-laccase chimera were encapsulated within an alginate hydrogel, and the resulting engineered living material released sufficient enzyme to decolorize a dye pollutant in its surroundings. These findings streamline recombinant protein production in cyanobacteria and facilitate environmental interactions with cell products, expanding the potential applications of both free-living and material-encapsulated cyanobacteria.

Keywords: bioremediation; cyanobacteria; engineered living materials; protein secretion; signal peptides

DOI: https://doi.org/10.1073/pnas.2624792123

Colloids Surf B Biointerfaces. 2026 Oct 15. pii: S0927-7765(26)00851-9. [Epub ahead of print]270 116263

Engineering electrospun living nanofibers for oral therapeutics.

M N Ferrace, F Montini-Ballarin, G Rivero, J Bruyère, H Cabral, D I Cattoni, G A Abraham.

Living nanofibers (LNs) are biohybrid fibrous materials that integrate living microorganisms within electrospun polymeric architectures, preserving their biological activity and enabling programmed therapeutic action at the target site. Their rational design requires integrating materials science and process engineering with biological constraints, using architectures that decouple and coordinate the protection, release, and host interaction of the encapsulated microorganisms. LNs can further incorporate conventional therapeutic agents, including small-molecule drugs, peptides, and biologics, enabling combinatorial strategies in which living or bioactive components act synergistically with drug-based therapies. In this review, we discuss the fundamental principles guiding LNs design, including material and strain selection, electrospinning-based fabrication techniques, and approaches to preserve viability and functionality during processing and storage. We then examine the physicochemical and biological barriers to oral delivery, including gastric acidity, enzymatic degradation and mucosal and epithelial barriers, and highlight emerging strategies to overcome these limitations. Finally, we discuss current and prospective applications of LNs in gastrointestinal and oncological therapies, emphasizing current capabilities for site-directed release and the requirements that remain for responsive therapeutic behaviour.

Keywords: Bacteriotherapy; Biofabrication; Electrospinning; Nanofibers; Probiotics

DOI: https://doi.org/10.1016/j.colsurfb.2026.116263

Biotechnol J. 2026 Oct;21(10): e70310

Synthetic Biology Toolbox for Lactiplantibacillus plantarum: Engineering Next-Generation Probiotic Cell Factories.

Peiyun Su, Yulin Ma, Qian Nie, Tingxing Yang, Tao Wei.

Lactiplantibacillus plantarum has emerged as a promising synthetic biology chassis owing to its food-grade safety, probiotic properties, and adaptability to the gastrointestinal tract. Its engineering potential has expanded rapidly in live biotherapeutics, functional foods, biosensors, and animal health. However, its broader application is still limited by the lack of standardized genetic parts, versatile engineering tools, and industrially compatible chassis. This review summarizes recent advances in the synthetic biology toolbox of L. plantarum, including expression elements, genome engineering strategies, and representative applications. Particular emphasis is placed on the development of genetic components, conventional and CRISPR-based engineering technologies, and their applications in medicine, food biotechnology, biosensing, and animal nutrition. Current challenges and future opportunities, including standardized genetic parts, precision genome engineering, AI-assisted strain design, and high-performance chassis development, are also discussed. This review provides a comprehensive framework for developing next-generation programmable probiotic cell factories based on L. plantarum.

Keywords: Lactiplantibacillus plantarum; expression systems; gene editing tools; synthetic biology

DOI: https://doi.org/10.1002/biot.70310

Folia Microbiol (Praha). 2026 Oct 17.

Smart carrier-engineered microbial consortia for precision nutrient delivery in next- generation biofertilizers.

Sangeeta Chhetri, Sharon Nagpal.

With the increasing demand for sustainable agricultural productivity under a changing climate, the advancement of new-generation biofertilizers specifically based on engineered microbial consortia and smart delivery systems has become an integral part of agricultural activities. The development of next-generation biofertilizers based on engineered microbial consortia and smart delivery systems has emerged as a promising approach for sustainable nutrient management. The drawback associated with conventional microbial inoculants is their inconsistent field performance, which is attributed to the limited viability of microbes under field conditions, environmental factors, and poorly controlled nutrient release. Recent advances in synthetic biology, microbial engineering, and biomaterials have enabled the development of beneficial microbial consortia for agricultural applications. Combining these engineered microbial consortia with advanced carrier technologies, including hydrogels derived from biopolymers, nanomaterials, bio-based polymers, and responsive encapsulation matrices, can enhance microbial protection, controlled delivery, and persistence in soil. The application of methods for encapsulating microorganisms, such as spray drying, ionic gelation, extrusion, and microfluidic encapsulation, can regulate microbial release and may improve microbial survival and rhizosphere colonization. Smart carrier systems are designed to protect microbial cells and regulate their release in response to environmental and plant-associated conditions, thereby supporting more controlled microbial activity and nutrient delivery. This review examines recent developments in engineered microbial consortia, advanced encapsulation techniques, and controlled-release systems for improving nutrient-use efficiency in next-generation biofertilizer formulations.

Keywords: Controlled release; Encapsulation technology; Microbial consortia; Next-generation biofertilizers; Nutrient use efficiency; Smart carrier system; Sustainable agriculture

DOI: https://doi.org/10.1007/s12223-026-01613-z