Magnetomechanical Detachment of Bacterial Biofilms Using Anisotropic Magnetic Iron Oxide Nanochains.

Bacterial biofilms attach to various surfaces and represent an important clinical and public health problem, as they are highly recalcitrant and are often associated with chronic, nonhealing diseases and healthcare-associated infections. Antibacterial agents are often not sufficient for their elimination and have to be combined with mechanical removal. Mechanical forces can be generated by actuating nonspherical (anisotropic) magnetically responsive nanoparticles in a rotating magnetic field.

Development of cellulose-degrading lactic acid bacterium Lactococcus cremoris by genetic engineering.

Cellulose is one of the most abundant potential sources of carbon for sustainable microbial production of biochemicals. Lactic acid bacteria that produce a range of valuable metabolites are generally unable to grow on cellulose. Here, we aimed to develop the lactic acid bacteria Lactococcus cremoris for growth on cellulosic substrate.

Lactococcus lactis as a delivery system for surface displayed mimotopes of major peanut allergen Ara h 2.

The prevalence of peanut allergy has been steadily increasing over the last decades. Among food allergies it shows the highest severity and is potentially fatal. Allergen Ara h 2 stands out as the main elicitor of allergic reactions.

Lactic acid bacteria as microbial cell factories for the in vivo delivery of therapeutic proteins as secretable TAT fusion products.

Protein-based therapeutics have garnered increasing attention across various medical fields for their substantial benefits to human health. Existing strategies for intracellular protein delivery, such as cell-penetrating peptide (CPP)-based approaches, including the transactivator of transcription (TAT) peptide, have shown promising results but also present limitations, particularly due to the need for recombinant protein manufacturing and daily invasive administration. To overcome such hindrances and develop delivery tools that are able to foster the production of the protein directly inside the body of patients, we engineered the lactic acid bacterium, Lactococcus lactis, to express and secrete TATκ-GFP protein.

Synergies between cellulosomal and non-cellulosomal bacterial cellulases complexed in designer cellulosomes.

The development of cellulases cocktails to efficiently solubilize cellulose for biotechnological applications is a constant demand. Cellulases exhibit diverse mode of action, operating in the free state or in multi-enzyme complexes (cellulosomes), and potentially acting synergistically for a higher efficiency. The efficacy of three non-cellulosomal bacterial cellulases originating from three different species that naturally act in the free state: Cel9A from Lachnoclostridium phytofermentans, Cel5H from Saccharophagus degradans and two variants of Cel5I from Ruminiclostridium cellulolyticum, were evaluated in bifunctional or trifunctional designer cellulosomes, and in combination with the most efficient pair of cellulases reported in R.

Biofilm Assessment by NanoLuc Luciferase Assay.

, a widespread pathogen found in birds and mammals, poses a significant risk for zoonosis worldwide despite its susceptibility to environmental and food-processing stressors. One of its main survival mechanisms is the formation of biofilms that can withstand various food-processing stressors, which is why efficient methods for assessing biofilms are crucial. Existing methods, including the classical culture-based plate counting method, biomass-staining methods (e.

Viability of potential probiotics incorporated into nanofibers: Influence of genera, storage conditions, stabilizers and their solid-state.

Electrospun nanofibers have emerged as a promising platform for probiotic delivery, with bacterial preservation posing a significant challenge in formulation design. This study examined the preservation of bacteria in various poly(ethylene oxide)-based nanofiber formulations and the solid-state behaviour of the excipients after electrospinning and during 24 weeks of storage under different conditions. Nanofiber formulations were loaded with bacteria from three different genera (oral cavity isolates Staphylococcus 26.

Incorporation of recombinant proteins into extracellular vesicles by Lactococcus cremoris.

Extracellular vesicles (EVs) are nanosized lipid bilayer particles released by various cellular organisms that carry an array of bioactive molecules. EVs have diagnostic potential, as they play a role in intercellular interspecies communication, and could be applied in drug delivery. In contrast to mammalian cell-derived EVs, the study of EVs from bacteria, particularly Gram-positive bacteria, received less research attention.

Holistic monitoring of Campylobacter jejuni biofilms with NanoLuc bioluminescence.

Campylobacter jejuni, a major cause of foodborne zoonotic infections worldwide, shows a paradoxical ability to survive despite its susceptibility to environmental and food-processing stressors. This resilience is likely due to the bacterium entering a viable but non-culturable state, often within biofilms, or even initiating biofilm formation as a survival strategy. This study presents an innovative application of NanoLuc bioluminescence to accurately monitor the development of C.

Modified vaginal lactobacilli expressing fluorescent and luminescent proteins for more effective monitoring of their release from nanofibers, safety and cell adhesion.

Electrospun nanofibers offer a highly promising platform for the delivery of vaginal lactobacilli, providing an innovative approach to preventing and treating vaginal infections. To advance the application of nanofibers for the delivery of lactobacilli, tools for studying their safety and efficacy in vitro need to be established. In this study, fluorescent (mCherry and GFP) and luminescent (NanoLuc luciferase) proteins were expressed in three vaginal lactobacilli (Lactobacillus crispatus, Lactobacillus gasseri and Lactobacillus jensenii) and a control Lactiplantibacillus plantarum with the aim to use this technology for close tracking of lactobacilli release from nanofibers and their adhesion on epithelial cells.

New treatment approaches for infections: alternatives to antibiotics and fecal microbiota transplantation.

causes a range of debilitating intestinal symptoms that may be fatal. It is particularly problematic as a hospital-acquired infection, causing significant costs to the health care system. Antibiotics, such as vancomycin and fidaxomicin, are still the drugs of choice for infections, but their effectiveness is limited, and microbial interventions are emerging as a new treatment option.

Secretion and surface display of binders of IL-23/IL-17 cytokines and their receptors in Lactococcus lactis as a therapeutic approach against inflammation.

The cytokine IL-23 activates the IL-23 receptor (IL-23R) and stimulates the differentiation of naïve T helper (Th) cells into a Th17 cell population that secretes inflammatory cytokines and chemokines. This IL-23/Th17 proinflammatory axis drives inflammation in Crohn's disease and ulcerative colitis and represents a therapeutic target of monoclonal antibodies. Non-immunoglobulin binding proteins based on the Streptococcus albumin-binding domain (ABD) provide a small protein alternative to monoclonal antibodies.

Targeting IL-6 by engineered Lactococcus lactis via surface-displayed affibody.

Background: Dysregulated production of interleukin (IL)-6 is implicated in the pathology of inflammatory bowel disease (IBD). Neutralization of IL-6 in the gut by safe probiotic bacteria may help alleviate intestinal inflammation. Here, we developed Lactococcus lactis with potent and selective IL-6 binding activity by displaying IL-6-specific affibody on its surface.

Influence of Excipient Composition on Survival of Vaginal Lactobacilli in Electrospun Nanofibers.

The lack of appropriate delivery systems hinders the use of probiotics in the treatment of vaginal infections. Therefore, the development of a new delivery system for the local administration of vaginal probiotics is necessary. In this study, we selected three vaginal lactobacilli, i.

Biofilm Assay Using NanoLuc Luciferase.

Biofilms serve as a bacterial survival strategy, allowing bacteria to persist under adverse environmental conditions. The non-pathogenic Listeria innocua is used as a surrogate organism for the foodborne pathogen Listeria monocytogenes, because they share genetic and physiological similarities and can be used in a Biosafety Level 1 laboratory. Several methods are used to evaluate biofilms, including different approaches to determine biofilm biomass or culturability, viability, metabolic activity, or other microbial community properties.

Dual Functionalized Shows Tumor Antigen Targeting and Cytokine Binding .

Pro-inflammatory cytokines play an important role in the development and progression of colorectal cancer (CRC). Tumor-targeting bacteria that can capture pro-inflammatory cytokines in the tumor microenvironment and thus block their tumor-promoting effects might provide clinical benefits in inflammation-associated CRC. The aim of this study was to develop bacteria with dual functionality for selective delivery of cytokine-binding proteins to the tumor by targeting specific receptors on cancer cells.

Introduction of Modified BglBrick System in for Straightforward Assembly of Multiple Gene Cassettes.

Genetic modification of lactic acid bacteria is an evolving and highly relevant field of research that allows the engineered bacteria to be equipped with the desired functions through the controlled expression of the recombinant protein. Novel genetic engineering techniques offer the advantage of being faster, easier and more efficient in incorporating modifications to the original bacterial strain. Here, we have developed a modified BglBrick system, originally introduced in and optimized it for the lactic acid bacterium .

Engineering of Vaginal Lactobacilli to Express Fluorescent Proteins Enables the Analysis of Their Mixture in Nanofibers.

Lactobacilli are a promising natural tool against vaginal dysbiosis and infections. However, new local delivery systems and additional knowledge about their distribution and mechanism of action would contribute to the development of effective medicine. This will be facilitated by the introduction of the techniques for effective, inexpensive, and real-time tracking of these probiotics following their release.

Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour-antigen binding proteins.

Development of targeted treatment for colorectal cancer is crucial to avoid side effects. To harness the possibilities offered by microbiome engineering, we prepared safe multifunctional cancer cell-targeting bacteria Lactococcus lactis. They displayed, on their surface, binding proteins for cancer-associated transmembrane receptors epithelial cell adhesion molecule (EpCAM) and human epidermal growth factor receptor 2 (HER2) and co-expressed an infrared fluorescent protein for imaging.

Expression of NanoLuc Luciferase in for Development of Biofilm Assay.

Studies of biofilm formation by bacteria are crucial for understanding bacterial resistance and for development of novel antibacterial strategies. We have developed a new bioluminescence biofilm assay for , which is considered a non-pathogenic surrogate for was transformed with a plasmid for inducible expression of NanoLuc luciferase (Nluc). Concentration-dependent bioluminescence signals were obtained over a concentration range of more than three log units.

Lectin-Mediated Binding of Engineered to Cancer Cells.

Lectins have been increasingly utilized as carriers for targeted drug delivery based on their specific binding to glycans located on mammalian cells. This study employed two lectins, B subunit of bacterial Shiga holotoxin (Stx1B) and fungal lectin (CNL), for surface display on the lactic acid bacterium . The specific adhesion of these engineered, lectin-displaying to cancer cells was evaluated.

The Influence of Probiotics on the Firmicutes/Bacteroidetes Ratio in the Treatment of Obesity and Inflammatory Bowel disease.

The two most important bacterial phyla in the gastrointestinal tract, Firmicutes and Bacteroidetes, have gained much attention in recent years. The Firmicutes/Bacteroidetes (F/B) ratio is widely accepted to have an important influence in maintaining normal intestinal homeostasis. Increased or decreased F/B ratio is regarded as dysbiosis, whereby the former is usually observed with obesity, and the latter with inflammatory bowel disease (IBD).

Electrospun Nanofibers as Carriers of Microorganisms, Stem Cells, Proteins, and Nucleic Acids in Therapeutic and Other Applications.

Electrospinning is a technique that uses polymer solutions and strong electric fields to produce nano-sized fibers that have wide-ranging applications. We present here an overview of the use of electrospinning to incorporate biological products into nanofibers, including microorganisms, cells, proteins, and nucleic acids. Although the conditions used during electrospinning limit the already problematic viability/stability of such biological products, their effective incorporation into nanofibers has been shown to be feasible.

Safety Aspects of Genetically Modified Lactic Acid Bacteria.

Lactic acid bacteria (LAB) have a long history of use in the food industry. Some species are part of the normal human microbiota and have beneficial properties for human health. Their long-standing use and considerable biotechnological potential have led to the development of various systems for their engineering.

Effects of Electrospinning on the Viability of Ten Species of Lactic Acid Bacteria in Poly(Ethylene Oxide) Nanofibers.

Lactic acid bacteria can have beneficial health effects and be used for the treatment of various diseases. However, there remains the challenge of encapsulating probiotics into delivery systems with a high viability and encapsulation efficacy. The electrospinning of bacteria is a novel and little-studied method, and further investigation of its promising potential is needed.