Optogenetic Engineered Living Materials.

Engineered living materials (ELM) is a new frontier in materials research that uses living microorganisms to augment nonliving materials with lifelike capabilities, such as responding to external stimuli. This is achieved by genetically programming the microorganisms in an ELM with stimulus-sensing modules. A popular stimulus to remotely control various ELM functions is light, which has been realized thanks to optogenetics.

Thermo-amplifier circuit in probiotic E. coli for stringently temperature-controlled release of a novel antibiotic.

Peptide drugs have seen rapid advancement in biopharmaceutical development, with over 80 candidates approved globally. Despite their therapeutic potential, the clinical translation of peptide drugs is hampered by challenges in production yields and stability. Engineered bacterial therapeutics is a unique approach being explored to overcome these issues by using bacteria to produce and deliver therapeutic compounds at the body site of use.

Engineered bacterial therapeutics with material solutions.

Recent advances in engineered bacterial therapeutics underscore their potential in treating diseases via targeted, live interventions. Despite their promising performance in early clinical phases, no engineered therapeutic bacteria have yet received approval, primarily due to challenges in proving efficacy while ensuring biosafety. Material science innovations, particularly the encapsulation of bacteria within hydrogels, present a promising avenue to enhance bacterial survival, efficacy, and safety in therapeutic applications.

Sustainable protein regeneration in encapsulated materials.

Zhu et al. introduce MELG (materials engineered by living grafting), combining engineered microbes with non-living scaffolds for functional protein regeneration within. These MELGs can be used for long-term controlled release, enzyme-mediated biocatalysis, and DNA purification.

Discovery of a high-performance phage-derived promoter/repressor system for probiotic lactobacillus engineering.

Background: The Lactobacillaceae family comprises many species of great importance for the food and healthcare industries, with numerous strains identified as beneficial for humans and used as probiotics. Hence, there is a growing interest in engineering these probiotic bacteria as live biotherapeutics for animals and humans. However, the genetic parts needed to regulate gene expression in these bacteria remain limited compared to model bacteria like E.

Rheological behavior of Pluronic/Pluronic diacrylate hydrogels used for bacteria encapsulation in engineered living materials.

Pluronic (Plu) hydrogels mixed with variable fractions of Pluronic diacrylate (PluDA) have become popular matrices to encapsulate bacteria and control their growth in engineered living materials. Here we study the rheological response of 30 wt% Plu/PluDA hydrogels with PluDA fraction between 0 and 1. We quantify the range of viscoelastic properties that can be covered in this system by varying in the PluDA fraction.

Engineered living materials for the conversion of a low-cost food-grade precursor to a high-value flavonoid.

Microbial biofactories allow the upscaled production of high-value compounds in biotechnological processes. This is particularly advantageous for compounds like flavonoids that promote better health through their antioxidant, anti-bacterial, anti-cancer and other beneficial effects but are produced in small quantities in their natural plant-based hosts. Bacteria like have been genetically modified with enzyme cascades to produce flavonoids like naringenin and pinocembrin from coumaric or cinnamic acid.

Expanding the genetic programmability of Lactiplantibacillus plantarum.

Lactobacilli are ubiquitous in nature and symbiotically provide health benefits for countless organisms including humans, animals and plants. They are vital for the fermented food industry and are being extensively explored for healthcare applications. For all these reasons, there is considerable interest in enhancing and controlling their capabilities through the engineering of genetic modules and circuits.

Retraction: Photoactivatable Hsp47: A Tool to Regulate Collagen Secretion and Assembly.

Adv. Sci. 2019, 6, 1801982 DOI: 10.

In vitro evaluation of immune responses to bacterial hydrogels for the development of living therapeutic materials.

In living therapeutic materials (LTMs), organisms genetically programmed to produce and deliver drugs are encapsulated in porous matrices acting as physical barriers between the therapeutic organisms and the host cells. LTMs consisting of engineered E. coli encapsulated in Pluronic F127-based hydrogels have been frequently used in LTM designs but their immunogenicity has not been tested.

Plasmonic stimulation of gold nanorods for the photothermal control of engineered living materials.

Engineered living materials (ELMs) encapsulate microorganisms within polymeric matrices for biosensing, drug delivery, capturing viruses, and bioremediation. It is often desirable to control their function remotely and in real time and so the microorganisms are often genetically engineered to respond to external stimuli. Here, we combine thermogenetically engineered microorganisms with inorganic nanostructures to sensitize an ELM to near infrared light.

In vitro assembly of plasmid DNA for direct cloning in Lactiplantibacillus plantarum WCSF1.

Lactobacilli are gram-positive bacteria that are growing in importance for the healthcare industry and genetically engineering them as living therapeutics is highly sought after. However, progress in this field is hindered since most strains are difficult to genetically manipulate, partly due to their complex and thick cell walls limiting our capability to transform them with exogenous DNA. To overcome this, large amounts of DNA (>1 μg) are normally required to successfully transform these bacteria.

Novel genetic modules encoding high-level antibiotic-free protein expression in probiotic lactobacilli.

Lactobacilli are ubiquitous in nature, often beneficially associated with animals as commensals and probiotics, and are extensively used in food fermentation. Due to this close-knit association, there is considerable interest to engineer them for healthcare applications in both humans and animals, for which high-performance and versatile genetic parts are greatly desired. For the first time, we describe two genetic modules in Lactiplantibacillus plantarum that achieve high-level gene expression using plasmids that can be retained without antibiotics, bacteriocins or genomic manipulations.

Encapsulation of bacteria in bilayer Pluronic thin film hydrogels: A safe format for engineered living materials.

In engineered living materials (ELMs) non-living matrices encapsulate microorganisms to acquire capabilities like sensing or biosynthesis. The confinement of the organisms to the matrix and the prevention of overgrowth and escape during the lifetime of the material is necessary for the application of ELMs into real devices. In this study, a bilayer thin film hydrogel of Pluronic F127 and Pluronic F127 acrylate polymers supported on a solid substrate is introduced.

Regulating Bacterial Behavior within Hydrogels of Tunable Viscoelasticity.

Engineered living materials (ELMs) are a new class of materials in which living organism incorporated into diffusive matrices uptake a fundamental role in material's composition and function. Understanding how the spatial confinement in 3D can regulate the behavior of the embedded cells is crucial to design and predict ELM's function, minimize their environmental impact and facilitate their translation into applied materials. This study investigates the growth and metabolic activity of bacteria within an associative hydrogel network (Pluronic-based) with mechanical properties that can be tuned by introducing a variable degree of acrylate crosslinks.

Exogenous supply of Hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro.

Background: Collagen is a structural protein that provides mechanical stability and defined architectures to skin. In collagen-based skin disorders this stability is lost, either due to mutations in collagens or in the chaperones involved in collagen assembly. This leads to chronic wounds, skin fragility, and blistering.

Photoactivatable Hsp47: A Tool to Regulate Collagen Secretion and Assembly.

Collagen is the most abundant structural protein in mammals and is crucial for the mechanical integrity of tissues. Hsp47, an endoplasmic reticulum resident collagen-specific chaperone, is involved in collagen biosynthesis and plays a fundamental role in the folding, stability, and intracellular transport of procollagen triple helices. This work reports on a photoactivatable derivative of Hsp47 that allows regulation of collagen biosynthesis within mammalian cells using light.

Optoregulated Protein Release from an Engineered Living Material.

Developing materials to encapsulate and deliver functional proteins inside the body is a challenging yet rewarding task for therapeutic purposes. High production costs, mostly associated with the purification process, short-term stability in vivo, and controlled and prolonged release are major hurdles for the clinical application of protein-based biopharmaceuticals. In an attempt to overcome these hurdles, herein, the possibility of incorporating bacteria as protein factories into a material and externally controlling protein release using optogenetics is demonstrated.

Optoregulated Drug Release from an Engineered Living Material: Self-Replenishing Drug Depots for Long-Term, Light-Regulated Delivery.

On-demand and long-term delivery of drugs are common requirements in many therapeutic applications, not easy to be solved with available smart polymers for drug encapsulation. This work presents a fundamentally different concept to address such scenarios using a self-replenishing and optogenetically controlled living material. It consists of a hydrogel containing an active endotoxin-free Escherichia coli strain.

Toward Light-Regulated Living Biomaterials.

Living materials are an emergent material class, infused with the productive, adaptive, and regenerative properties of living organisms. Property regulation in living materials requires encoding responsive units in the living components to allow external manipulation of their function. Here, an optoregulated -based living biomaterial that can be externally addressed using light to interact with mammalian cells is demonstrated.

Cell Adhesion on RGD-Displaying Knottins with Varying Numbers of Tryptophan Amino Acids to Tune the Affinity for Assembly on Cucurbit[8]uril Surfaces.

Cell adhesion is studied on multivalent knottins, displaying RGD ligands with a high affinity for integrin receptors, that are assembled on CB[8]-methylviologen-modified surfaces. The multivalency in the knottins stems from the number of tryptophan amino acid moieties, between 0 and 4, that can form a heteroternary complex with cucurbit[8]uril (CB[8]) and surface-tethered methylviologen (MV). The binding affinity of the knottins with CB[8] and MV surfaces was evaluated using surface plasmon resonance spectroscopy.

Cell Adhesion on Dynamic Supramolecular Surfaces Probed by Fluid Force Microscopy-Based Single-Cell Force Spectroscopy.

Biomimetic and stimuli-responsive cell-material interfaces are actively being developed to study and control various cell-dynamics phenomena. Since cells naturally reside in the highly dynamic and complex environment of the extracellular matrix, attempts are being made to replicate these conditions in synthetic biomaterials. Supramolecular chemistry, dealing with noncovalent interactions, has recently provided possibilities to incorporate such dynamicity and responsiveness in various types of architectures.

Agglutination of bacteria using polyvalent nanoparticles of aggregation-induced emissive thiophthalonitrile dyes.

A novel class of aggregation-induced emissive bis(phenylthio)phthalonitrile dyes were synthesized. These dyes assembled into nanoparticles that were equipped with mannose units. The nanoparticles underwent selective interactions with lectins and bacteria.

Photoresponsive Cucurbit[8]uril-Mediated Adhesion of Bacteria on Supported Lipid Bilayers.

In this work, the development of a photoresponsive platform for the presentation of bioactive ligands to study receptor-ligand interactions has been described. For this purpose, supramolecular host-guest chemistry and supported lipid bilayers (SLBs) have been combined in a microfluidic device. Quartz crystal microbalance with dissipation monitoring (QCM-D) studies on methyl viologen (MV)-functionalized oligo ethylene glycol-based self-assembled monolayers, gel and liquid-state SLBs have been compared for their nonfouling properties in the case of ConA and bacteria.