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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 |
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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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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.</description><identifier>ISSN: 1751-7915</identifier><identifier>EISSN: 1751-7915</identifier><identifier>DOI: 10.1111/1751-7915.13572</identifier><identifier>PMID: 32348028</identifier><language>eng</language><publisher>United States: John Wiley & Sons, Inc</publisher><subject>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</subject><ispartof>Microbial biotechnology, 2020-07, Vol.13 (4), p.1162-1178</ispartof><rights>2020 The Authors. published by John Wiley & Sons Ltd and Society for Applied Microbiology.</rights><rights>2020 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.</rights><rights>2020. This work is published under http://creativecommons.org/licenses/by-nc-nd/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c6672-1a48022e34caed011550790fa5992821c8e3aaeaa1da88089232341cc54075993</citedby><cites>FETCH-LOGICAL-c6672-1a48022e34caed011550790fa5992821c8e3aaeaa1da88089232341cc54075993</cites><orcidid>0000-0002-4802-8518</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/2408476229/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2408476229?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,11541,25731,27901,27902,36989,36990,44566,46027,46451,53766,53768,74869</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32348028$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hirayama, Satoru</creatorcontrib><creatorcontrib>Nakao, Ryoma</creatorcontrib><title>Glycine significantly enhances bacterial membrane vesicle production: a powerful approach for isolation of LPS‐reduced membrane vesicles of probiotic Escherichia coli</title><title>Microbial biotechnology</title><addtitle>Microb Biotechnol</addtitle><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.</description><subject>Adjuvanticity</subject><subject>Animals</subject><subject>Bacteria</subject><subject>Bacterial vesicles</subject><subject>Cell culture</subject><subject>Deficient mutant</subject><subject>E coli</subject><subject>Endotoxins</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Flagella</subject><subject>Glycine</subject><subject>Immune response</subject><subject>Immunomodulation</subject><subject>Lipids</subject><subject>Lipopolysaccharides</subject><subject>Lysis</subject><subject>Macrophages</subject><subject>Membrane proteins</subject><subject>Membrane vesicles</subject><subject>Membranes</subject><subject>Mice</subject><subject>Morphology</subject><subject>Nanoparticles</subject><subject>Probiotics</subject><subject>Proteins</subject><subject>Stationary phase</subject><subject>Supplements</subject><subject>Vaccines</subject><subject>Vesicles</subject><issn>1751-7915</issn><issn>1751-7915</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNqFks9u1DAQxiMEomXhzA1Z4sJlW_9J4oQDElSlVFoEEuVsTezxritvvNhJq73xCDwGz8WT4HTLqoUDvtia-c3nmdFXFM8ZPWL5HDNZsblsWXXERCX5g-JwH3l4531QPEnpktKa0oo_Lg4EF2VDeXNY_DzzW-16JMkte2edhn7wW4L9CnqNiXSgB4wOPFnjuouQyStMTnskmxjMqAcX-tcEyCZcY7SjJ7DJCdArYkMkLgUPE0KCJYvPX359_xExV6H5Ry9NSC7tXBicJqdJr_LHeuWA6ODd0-KRBZ_w2e09K76-P704-TBffDo7P3m7mOu6lnzOYBqMoyg1oKGMVRWVLbVQtS1vONMNCgAEYAaahjYtn3bBtK5KKjMjZsX5TtcEuFSb6NYQtyqAUzeBEJcK4jA1rDorOo1WlrSypdC2E4KjNcZAC0aaLmu92Wltxm6NRmM_RPD3RO9nerdSy3ClJK_LqbVZ8epWIIZvI6ZBrV3S6H3eWxiT4qKtBZWS04y-_Au9DGPs86oUL2lTyprzabrjHaVjSCmi3TfDqJocpSbPqMkz6sZRueLF3Rn2_B8LZaDeAdfO4_Z_eurjuwu-U_4NM7XbKQ</recordid><startdate>202007</startdate><enddate>202007</enddate><creator>Hirayama, Satoru</creator><creator>Nakao, Ryoma</creator><general>John Wiley & Sons, Inc</general><general>John Wiley and Sons Inc</general><general>Wiley</general><scope>24P</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QO</scope><scope>7T7</scope><scope>7X7</scope><scope>7XB</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0S</scope><scope>M7P</scope><scope>M7S</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-4802-8518</orcidid></search><sort><creationdate>202007</creationdate><title>Glycine significantly enhances bacterial membrane vesicle production: a powerful approach for isolation of LPS‐reduced membrane vesicles of probiotic Escherichia coli</title><author>Hirayama, Satoru ; 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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.</abstract><cop>United States</cop><pub>John Wiley & Sons, Inc</pub><pmid>32348028</pmid><doi>10.1111/1751-7915.13572</doi><tpages>17</tpages><orcidid>https://orcid.org/0000-0002-4802-8518</orcidid><oa>free_for_read</oa></addata></record> |
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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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