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Glycine significantly enhances bacterial membrane vesicle production: a powerful approach for isolation of LPS‐reduced membrane vesicles of probiotic Escherichia coli

Summary Bacterial membrane vesicles (MVs) have attracted strong interest in recent years as novel nanoparticle delivery platforms. Glycine is known to induce morphological changes in the outer layer of bacteria. We report here that glycine dramatically facilitates MV production in a flagella‐deficie...

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Published in:Microbial biotechnology 2020-07, Vol.13 (4), p.1162-1178
Main Authors: Hirayama, Satoru, Nakao, Ryoma
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description Summary Bacterial membrane vesicles (MVs) have attracted strong interest in recent years as novel nanoparticle delivery platforms. Glycine is known to induce morphological changes in the outer layer of bacteria. We report here that glycine dramatically facilitates MV production in a flagella‐deficient mutant of the non‐pathogenic probiotic Escherichia coli strain Nissle 1917. Supplementation of culture medium with 1.0% glycine induced cell deformation at the early exponential phase, eventually followed by quasi‐lysis during the late exponential to stationary phase. Glycine supplementation also significantly increased the number of MVs with enlarged particle size and altered the protein profile with an increase in the inner membrane and cytoplasmic protein contents as compared to non‐induced MVs. Of note, the endotoxin activity of glycine‐induced MVs was approximately eightfold or sixfold lower than that of non‐induced MVs when compared at equal protein or lipid concentrations respectively. Nevertheless, glycine‐induced MVs efficiently induced both immune responses in a mouse macrophage‐like cell line and adjuvanticity in an intranasal vaccine mouse model, comparable to those of non‐induced MVs. We propose that the present method of inducing MV production with glycine can be used for emerging biotechnological applications of MVs that have immunomodulatory activities, while dramatically reducing the presence of endotoxins. Physiologically excessive amount of glycine significantly enhanced MV production from a flagella‐deficient clone of probiotic Escherichia coli strain Nissle 1917. The glycine‐induced MVs could elicit strong immune responses, while dramatically reducing the presence of endotoxins.
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subjects Adjuvanticity
Animals
Bacteria
Bacterial vesicles
Cell culture
Deficient mutant
E coli
Endotoxins
Escherichia coli
Escherichia coli - genetics
Flagella
Glycine
Immune response
Immunomodulation
Lipids
Lipopolysaccharides
Lysis
Macrophages
Membrane proteins
Membrane vesicles
Membranes
Mice
Morphology
Nanoparticles
Probiotics
Proteins
Stationary phase
Supplements
Vaccines
Vesicles
title Glycine significantly enhances bacterial membrane vesicle production: a powerful approach for isolation of LPS‐reduced membrane vesicles of probiotic Escherichia coli
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