Quantifying and engineering probiotic bacterial adhesion to mucus

Mucus in the gastrointestinal (GI) tract is the primary point-of-interaction between humans and their gut microbiota. This not only intimates that mucus ensures protection against endogenous and exogenous opportunists but provision for the human microbiota to reside and flourish. With the emergence of living therapeutics, engineered microbes can deliver and produce increasingly complex medicine, and controlling the mucoadhesive properties of different microbial chassis can dictate dose-response in a patient. Here we present a redesigned, in vitro, plate-based assay to measure the mucus adhesion of various probiotics. Cell-mucus interactions were isolated by immobilizing mucus to the plate surface. Binding parameters were derived for each probiotic strain by measuring cell adhesion over a wide range of cell concentrations, providing dose-dependent adhesion metrics. Surface proteins and cell components known to influence mucoadhesion were then heterologously expressed or altered in Lactococcus lactis MG1363 and E. coli Nissle 1917 to control mucus-binding capacity, avidity, and cooperativity.

• Publication year

  2019

• Venue

  bioRxiv

• Publication date

  2019-08-10

• Fields of study

  Biology, Chemistry, Engineering, Environmental Science, Medicine

• Identifiers
  DOI 10.1101/731505
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar

• No claims are published for this paper.

• No concepts are published for this paper.

• An engineered E. coli Nissle improves hyperammonemia and survival in mice and shows dose-dependent exposure in healthy humans

  2019cited by this paper
• Programmable bacteria induce durable tumor regression and systemic antitumor immunity

  2019cited by this paper
• Synthetic biology in probiotic lactic acid bacteria: At the frontier of living therapeutics.

  2018cited by this paper
• Gut Microbiota-Derived Tryptophan Metabolites Modulate Inflammatory Response in Hepatocytes and Macrophages

  2018cited by this paper
• Development of a synthetic live bacterial therapeutic for the human metabolic disease phenylketonuria

  2018cited by this paper
• Mucins and Their Role in Shaping the Functions of Mucus Barriers.

  2018cited by this paper
• Adaptive strategies of the candidate probiotic E. coli Nissle in the mammalian gut

  2018cited by this paper
• Synthetic Biology Approaches to Engineer Probiotics and Members of the Human Microbiota for Biomedical Applications.

  2018cited by this paper
• Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles

  2018cited by this paper
• Engineered probiotic Escherichia coli can eliminate and prevent Pseudomonas aeruginosa gut infection in animal models

  2017cited by this paper
• A review of dose-responses of probiotics in human studies.

  2017cited by this paper
• Mucin- and carbohydrate-stimulated adhesion and subproteome changes of the probiotic bacterium Lactobacillus acidophilus NCFM.

  2017cited by this paper
• Engineered bacteria can function in the mammalian gut long-term as live diagnostics of inflammation

  2017cited by this paper
• Innovative Methods and Applications in Mucoadhesion Research.

  2017cited by this paper
• Toward a genetic tool development pipeline for host-associated bacteria.

  2017cited by this paper
• The Interaction between Respiratory Pathogens and Mucus.

  2016cited by this paper
• Genetic engineering of probiotic Escherichia coli Nissle 1917 for clinical application

  2016cited by this paper
• The Gut-Brain Axis : Dietary, Probiotic, and Prebiotic Interventions on the Microbiota

  2016cited by this paper
• Food-Grade Organisms as Vaccine Biofactories and Oral Delivery Vehicles.

  2016cited by this paper
• Use of Atomic Force Microscopy to Study the Multi-Modular Interaction of Bacterial Adhesins to Mucins

  2016cited by this paper
• The Effect of Probiotic Plus Prebiotic Supplementation on the Tolerance and Efficacy of Helicobacter Pylori Eradication Quadruple Therapy: a Randomized Prospective Double Blind Controlled Trial

  2016cited by this paper
• Carbohydrate-binding specificities of potential probiotic Lactobacillus strains in porcine jejunal (IPEC-J2) cells and porcine mucin

  2016cited by this paper
• Use of probiotics in prevention and treatment of patients with Clostridium difficile infection.

  2016cited by this paper
• Ultrastructural Characterization of the Lower Motor System in a Mouse Model of Krabbe Disease

  2016cited by this paper
• Adhesion Properties of Lactic Acid Bacteria on Intestinal Mucin

  2016cited by this paper
• Biofilm Forming Lactobacillus: New Challenges for the Development of Probiotics

  2016cited by this paper
• The lectin-like protein 1 in Lactobacillus rhamnosus GR-1 mediates tissue-specific adherence to vaginal epithelium and inhibits urogenital pathogens

  2016cited by this paper
• Role and mechanisms of action of Escherichia coli Nissle 1917 in the maintenance of remission in ulcerative colitis patients: An update.

  2016cited by this paper
• A Review of Cell Adhesion Studies for Biomedical and Biological Applications

  2015cited by this paper
• The ecology of the microbiome: Networks, competition, and stability

  2015cited by this paper
• Milk fermented with a 15-lipoxygenase-1-producing Lactococcus lactis alleviates symptoms of colitis in a murine model.

  2015cited by this paper
• Programmable probiotics for detection of cancer in urine

  2015cited by this paper
• The gut microbiome in health and in disease

  2015cited by this paper
• Role of Flagella in Adhesion of Escherichia coli to Abiotic Surfaces.

  2015cited by this paper
• Bacteriocin-producing strains of Lactobacillus plantarum inhibit adhesion of Staphylococcus aureus to extracellular matrix: quantitative insight and implications in antibacterial therapy.

  2015cited by this paper
• Fate, activity, and impact of ingested bacteria within the human gut microbiota.

  2015cited by this paper
• Structural and molecular insights into novel surface‐exposed mucus adhesins from Lactobacillus reuteri human strains

  2014cited by this paper
• Heterologous Protein Secretion in Lactobacilli with Modified pSIP Vectors

  2014cited by this paper
• Functional genomics of lactic acid bacteria: from food to health

  2014cited by this paper
• Weissella confusa: problems with identification of an opportunistic pathogen that has been found in fermented foods and proposed as a probiotic

  2014cited by this paper
• Genetic basis for nitrate resistance in Desulfovibrio strains

  2014cited by this paper
• Role of Lactobacillus reuteri cell and mucus-binding protein A (CmbA) in adhesion to intestinal epithelial cells and mucus in vitro.

  2014cited by this paper
• Nutritional Basis for Colonization Resistance by Human Commensal Escherichia coli Strains HS and Nissle 1917 against E. coli O157:H7 in the Mouse Intestine

  2013cited by this paper
• Surface-associated MUC5B mucins promote protease activity in Lactobacillus fermentum biofilms

  2013cited by this paper
• Mucin-type O-glycans and their roles in intestinal homeostasis.

  2013cited by this paper
• Positive Outcomes Influence the Rate and Time to Publication, but Not the Impact Factor of Publications of Clinical Trial Results

  2013cited by this paper
• Unraveling the Role of Surface Mucus-Binding Protein and Pili in Muco-Adhesion of Lactococcus lactis

  2013cited by this paper
• Surface Display of N-Terminally Anchored Invasin by Lactobacillus plantarum Activates NF-κB in Monocytes

  2012cited by this paper
• The human microbiome: at the interface of health and disease

  2012cited by this paper
• Mucin biopolymers prevent bacterial aggregation by retaining cells in the free-swimming state.

  2012cited by this paper
• Escherichia coli Nissle 1917 Targets and Restrains Mouse B16 Melanoma and 4T1 Breast Tumors through Expression of Azurin Protein

  2012cited by this paper
• In vitro testing of commercial and potential probiotic lactic acid bacteria.

  2012cited by this paper
• How glycan metabolism shapes the human gut microbiota

  2012cited by this paper
• More than a marine propeller--the flagellum of the probiotic Escherichia coli strain Nissle 1917 is the major adhesin mediating binding to human mucus.

  2012cited by this paper
• Evaluation of the functional potential of Weissella and Lactobacillus isolates obtained from Nigerian traditional fermented foods and cow's intestine.

  2011cited by this paper
• The human gut microbiome: ecology and recent evolutionary changes.

  2011cited by this paper
• Diversification of the gut symbiont Lactobacillus reuteri as a result of host-driven evolution

  2010cited by this paper
• Probiotic Escherichia coli strain Nissle 1917 outcompetes intestinal pathogens during biofilm formation.

  2010cited by this paper
• Strain-specific diversity of mucus-binding proteins in the adhesion and aggregation properties of Lactobacillus reuteri.

  2010cited by this paper
• Survival of the probiotic Escherichia coli Nissle 1917 (EcN) in the gastrointestinal tract given in combination with oral mesalamine to healthy volunteers

  2010cited by this paper
• protein expression and preventing apoptosis in a murine model of colitis Probiotic mixture VSL#3 protects the epithelial barrier by maintaining tight junction

  2010cited by this paper
• Host-microbial symbiosis in the vertebrate gastrointestinal tract and the Lactobacillus reuteri paradigm

  2010cited by this paper
• Genome-wide analysis of signal peptide functionality in Lactobacillus plantarum WCFS1

  2009cited by this paper
• Comparative genomic analysis of Lactobacillus rhamnosus GG reveals pili containing a human- mucus binding protein

  2009cited by this paper
• Comparison of in vitro models to study bacterial adhesion to the intestinal epithelium

  2009cited by this paper
• A complex, but uniform O-glycosylation of the human MUC2 mucin from colonic biopsies analyzed by nanoLC/MSn.

  2009cited by this paper
• Probiotic mixture VSL#3 protects the epithelial barrier by maintaining tight junction protein expression and preventing apoptosis in a murine model of colitis.

  2009cited by this paper
• Identification of a Gene Cluster for the Biosynthesis of a Long, Galactose-Rich Exopolysaccharide in Lactobacillus rhamnosus GG and Functional Analysis of the Priming Glycosyltransferase

  2009cited by this paper
• Micro- and macrorheology of mucus.

  2009cited by this paper
• Escherichia coli Nissle 1917 for probiotic use in piglets: evidence for intestinal colonization

  2009cited by this paper
• Characterization of cellulose production in Escherichia coli Nissle 1917 and its biological consequences.

  2009cited by this paper
• F1C Fimbriae Play an Important Role in Biofilm Formation and Intestinal Colonization by the Escherichia coli Commensal Strain Nissle 1917

  2008cited by this paper
• Comparison of Carbon Nutrition for Pathogenic and Commensal Escherichia coli Strains in the Mouse Intestine

  2008cited by this paper
• The adhesion of putative probiotic lactobacilli to cultured epithelial cells and porcine intestinal mucus

  2008cited by this paper
• Strain‐ and matrix‐dependent adhesion of Lactobacillus plantarum is mediated by proteinaceous bacterial compounds

  2007cited by this paper
• Rheology of gastric mucin exhibits a pH-dependent sol-gel transition.

  2007cited by this paper
• Molecular structure and rheological properties of short-side-chain heavily glycosylated porcine stomach mucin.

  2007cited by this paper
• A Mucus Adhesion Promoting Protein, MapA, Mediates the Adhesion of Lactobacillus reuteri to Caco-2 Human Intestinal Epithelial Cells

  2006cited by this paper
• Comparative analysis of proteins with a mucus-binding domain found exclusively in lactic acid bacteria.

  2006cited by this paper
• A synthetic promoter library for constitutive gene expression in Lactobacillus plantarum.

  2006cited by this paper
• Carboxyfluorescein diacetate succinimidyl ester fluorescent dye for cell labeling.

  2005cited by this paper
• The cell surface of Lactobacillus reuteri ATCC 55730 highlighted by identification of 126 extracellular proteins from the genome sequence.

  2005cited by this paper
• Differential mucin expression in colon carcinoma HT‐29 clones with variable resistance to 5‐fluorouracil and methotrexate

  2004cited by this paper
• A New Assay Using Surface Plasmon Resonance (SPR) to Determine Binding of the Lactobacillus acidophilus Group to Human Colonic Mucin

  2004cited by this paper
• Intracellular Bacterial Biofilm-Like Pods in Urinary Tract Infections

  2003cited by this paper
• Purification and Characterization of a Surface Protein from Lactobacillus fermentum 104R That Binds to Porcine Small Intestinal Mucus and Gastric Mucin

  2002cited by this paper
• Investigation of the Functional Role of Ctp Proteins in the Cyanobacterium Synechocystis sp. PCC 6803

  2002cited by this paper
• A high-molecular-mass cell-surface protein from Lactobacillus reuteri 1063 adheres to mucus components.

  2002cited by this paper
• Addition of mucin to the growth medium triggers mucus-binding activity in different strains of Lactobacillus reuteri in vitro.

  2001cited by this paper
• Quantitative Approach in the Study of Adhesion of Lactic Acid Bacteria to Intestinal Cells and Their Competition with Enterobacteria

  2000cited by this paper
• Prophylaxis of pouchitis onset with probiotic therapy: A double‐blind, placebo‐controlled trial

  2000cited by this paper
• One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products.

  2000cited by this paper
• A collagen binding protein from Lactobacillus reuteri is part of an ABC transporter system?

  1996cited by this paper
• Cell surface protein receptors in oral streptococci.

  1994cited by this paper
• Purification of collagen-binding proteins ofLactobacillus reuteri NCIB 11951

  1994cited by this paper

• No citing papers are available for this paper.