Isolation and separation of the glycan strands from murein of Escherichia coli by reversed-phase high-performance liquid chromatography
The length distribution of the glycan strands in the murein (peptidoglycan) sacculus of Escherichia coli has been analyzed after solubilization of the murein by complete digestion with human serum amidase. The glycan strands released were separated according to length by reversed-phase HPLC on wide-...
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| Veröffentlicht in: | Analytical biochemistry 1990-10, Vol.190 (1), p.120-128 |
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| creator | Harz, Hartmann Burgdorf, Knut Höltje, Joachim-Volker |
| description | The length distribution of the glycan strands in the murein (peptidoglycan) sacculus of
Escherichia coli has been analyzed after solubilization of the murein by complete digestion with human serum amidase. The glycan strands released were separated according to length by reversed-phase HPLC on wide-pore Nucleosil 300 C
18 material at 50°C, employing a convex gradient from 5 to 11% acetonitrile. The length of the fractionated glycan strands, which carry a nonreducing 1,6-anhydromuramic acid as a natural end group, was calculated from the ratio of total to nonreducing terminal muramic acid residues. This was possible after complete hydrolysis of the isolated glycan strands by muramidase followed by separation of the released nonreducing and reducing di- and tetrasaccharides by reversed-phase HPLC on Hypersil C
18. The method established allows the separation of the glycan strands of murein, a poly-GlcNAc(β1-4)MurNAc-polysaccharide, up to a degree of polymerization of approximately 60. The predominant lengths of the glycan strands were 5 to 10 GlcNAc(β1-4)MurNAc disaccharide units. |
| doi_str_mv | 10.1016/0003-2697(90)90144-X |
| format | Article |
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18 material at 50°C, employing a convex gradient from 5 to 11% acetonitrile. The length of the fractionated glycan strands, which carry a nonreducing 1,6-anhydromuramic acid as a natural end group, was calculated from the ratio of total to nonreducing terminal muramic acid residues. This was possible after complete hydrolysis of the isolated glycan strands by muramidase followed by separation of the released nonreducing and reducing di- and tetrasaccharides by reversed-phase HPLC on Hypersil C
18. The method established allows the separation of the glycan strands of murein, a poly-GlcNAc(β1-4)MurNAc-polysaccharide, up to a degree of polymerization of approximately 60. The predominant lengths of the glycan strands were 5 to 10 GlcNAc(β1-4)MurNAc disaccharide units.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/0003-2697(90)90144-X</identifier><identifier>PMID: 2285138</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Acetylglucosamine - metabolism ; Amino Acid Sequence ; Carbohydrate Sequence ; Chromatography, High Pressure Liquid - methods ; Escherichia coli - analysis ; Humans ; Hydrogen-Ion Concentration ; Hydrolysis ; Molecular Sequence Data ; Molecular Weight ; Muramic Acids - metabolism ; Muramidase - metabolism ; Peptidoglycan - isolation & purification ; Peptidoglycan - metabolism ; Polysaccharides - isolation & purification ; Polysaccharides - metabolism</subject><ispartof>Analytical biochemistry, 1990-10, Vol.190 (1), p.120-128</ispartof><rights>1990</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c505t-848089930744f17510337b0bb2e965305e4c2b1163fb142de860379fa4b5bc5c3</citedby><cites>FETCH-LOGICAL-c505t-848089930744f17510337b0bb2e965305e4c2b1163fb142de860379fa4b5bc5c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0003-2697(90)90144-X$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2285138$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Harz, Hartmann</creatorcontrib><creatorcontrib>Burgdorf, Knut</creatorcontrib><creatorcontrib>Höltje, Joachim-Volker</creatorcontrib><title>Isolation and separation of the glycan strands from murein of Escherichia coli by reversed-phase high-performance liquid chromatography</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>The length distribution of the glycan strands in the murein (peptidoglycan) sacculus of
Escherichia coli has been analyzed after solubilization of the murein by complete digestion with human serum amidase. The glycan strands released were separated according to length by reversed-phase HPLC on wide-pore Nucleosil 300 C
18 material at 50°C, employing a convex gradient from 5 to 11% acetonitrile. The length of the fractionated glycan strands, which carry a nonreducing 1,6-anhydromuramic acid as a natural end group, was calculated from the ratio of total to nonreducing terminal muramic acid residues. This was possible after complete hydrolysis of the isolated glycan strands by muramidase followed by separation of the released nonreducing and reducing di- and tetrasaccharides by reversed-phase HPLC on Hypersil C
18. The method established allows the separation of the glycan strands of murein, a poly-GlcNAc(β1-4)MurNAc-polysaccharide, up to a degree of polymerization of approximately 60. The predominant lengths of the glycan strands were 5 to 10 GlcNAc(β1-4)MurNAc disaccharide units.</description><subject>Acetylglucosamine - metabolism</subject><subject>Amino Acid Sequence</subject><subject>Carbohydrate Sequence</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>Escherichia coli - analysis</subject><subject>Humans</subject><subject>Hydrogen-Ion Concentration</subject><subject>Hydrolysis</subject><subject>Molecular Sequence Data</subject><subject>Molecular Weight</subject><subject>Muramic Acids - metabolism</subject><subject>Muramidase - metabolism</subject><subject>Peptidoglycan - isolation & purification</subject><subject>Peptidoglycan - metabolism</subject><subject>Polysaccharides - isolation & purification</subject><subject>Polysaccharides - metabolism</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcFq3DAQhkVpSTdp36AFnUpzcDuyJMu6FEJI2kAglxRyE5I8XqvYliPZgX2Cvna82SXH9iSG_5tfMB8hnxh8Y8Cq7wDAi7LS6quGcw1MiOLhDdkw0FUBHPRbsnlF3pPTnP8AMCZkdUJOyrKWjNcb8vcmx97OIY7Ujg3NONl0GGNL5w7ptt95O9I8pzXPtE1xoMOSMLwQV9l3mILvgqU-9oG6HU34hCljU0ydzUi7sO2KCVMb02BHj7QPj0toqO_WKjvHbbJTt_tA3rW2z_jx-J6R39dX95e_itu7nzeXF7eFlyDnohY11FpzUEK0TEkGnCsHzpWoK8lBovClY6zirWOibLCugCvdWuGk89LzM_Ll0Dul-Lhgns0Qsse-tyPGJZsaSqGUgP-CTCpVAtMrKA6gTzHnhK2ZUhhs2hkGZi_K7C2YvQWjwbyIMg_r2udj_-IGbF6XjmbW_Mchx_UaTwGTyT7ger8mJPSzaWL49wfPdhejyQ</recordid><startdate>19901001</startdate><enddate>19901001</enddate><creator>Harz, Hartmann</creator><creator>Burgdorf, Knut</creator><creator>Höltje, Joachim-Volker</creator><general>Elsevier Inc</general><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>7QL</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>19901001</creationdate><title>Isolation and separation of the glycan strands from murein of Escherichia coli by reversed-phase high-performance liquid chromatography</title><author>Harz, Hartmann ; Burgdorf, Knut ; Höltje, Joachim-Volker</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c505t-848089930744f17510337b0bb2e965305e4c2b1163fb142de860379fa4b5bc5c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Acetylglucosamine - metabolism</topic><topic>Amino Acid Sequence</topic><topic>Carbohydrate Sequence</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Escherichia coli - analysis</topic><topic>Humans</topic><topic>Hydrogen-Ion Concentration</topic><topic>Hydrolysis</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>Muramic Acids - metabolism</topic><topic>Muramidase - metabolism</topic><topic>Peptidoglycan - isolation & purification</topic><topic>Peptidoglycan - metabolism</topic><topic>Polysaccharides - isolation & purification</topic><topic>Polysaccharides - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Harz, Hartmann</creatorcontrib><creatorcontrib>Burgdorf, Knut</creatorcontrib><creatorcontrib>Höltje, Joachim-Volker</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Harz, Hartmann</au><au>Burgdorf, Knut</au><au>Höltje, Joachim-Volker</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation and separation of the glycan strands from murein of Escherichia coli by reversed-phase high-performance liquid chromatography</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>1990-10-01</date><risdate>1990</risdate><volume>190</volume><issue>1</issue><spage>120</spage><epage>128</epage><pages>120-128</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><abstract>The length distribution of the glycan strands in the murein (peptidoglycan) sacculus of
Escherichia coli has been analyzed after solubilization of the murein by complete digestion with human serum amidase. The glycan strands released were separated according to length by reversed-phase HPLC on wide-pore Nucleosil 300 C
18 material at 50°C, employing a convex gradient from 5 to 11% acetonitrile. The length of the fractionated glycan strands, which carry a nonreducing 1,6-anhydromuramic acid as a natural end group, was calculated from the ratio of total to nonreducing terminal muramic acid residues. This was possible after complete hydrolysis of the isolated glycan strands by muramidase followed by separation of the released nonreducing and reducing di- and tetrasaccharides by reversed-phase HPLC on Hypersil C
18. The method established allows the separation of the glycan strands of murein, a poly-GlcNAc(β1-4)MurNAc-polysaccharide, up to a degree of polymerization of approximately 60. The predominant lengths of the glycan strands were 5 to 10 GlcNAc(β1-4)MurNAc disaccharide units.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>2285138</pmid><doi>10.1016/0003-2697(90)90144-X</doi><tpages>9</tpages></addata></record> |
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| ispartof | Analytical biochemistry, 1990-10, Vol.190 (1), p.120-128 |
| issn | 0003-2697 1096-0309 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Acetylglucosamine - metabolism Amino Acid Sequence Carbohydrate Sequence Chromatography, High Pressure Liquid - methods Escherichia coli - analysis Humans Hydrogen-Ion Concentration Hydrolysis Molecular Sequence Data Molecular Weight Muramic Acids - metabolism Muramidase - metabolism Peptidoglycan - isolation & purification Peptidoglycan - metabolism Polysaccharides - isolation & purification Polysaccharides - metabolism |
| title | Isolation and separation of the glycan strands from murein of Escherichia coli by reversed-phase high-performance liquid chromatography |
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