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concentration of polysaccharides and proteins in EPS of Pseudomonas putida and Aureobasidum pullulans as revealed by ¹³C CPMAS NMR spectroscopy
Extracellular polymeric substances were extracted from the bacterial strain Pseudomonas putida and the fungal species Aureobasidium pullulans using three different methods (formaldehyde-NaOH, ethylenediaminetetraacetic acid (EDTA) and cation-exchange-resin). The composition of the extracellular poly...
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Published in: | Applied microbiology and biotechnology 2009-11, Vol.85 (1), p.197-206 |
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description | Extracellular polymeric substances were extracted from the bacterial strain Pseudomonas putida and the fungal species Aureobasidium pullulans using three different methods (formaldehyde-NaOH, ethylenediaminetetraacetic acid (EDTA) and cation-exchange-resin). The composition of the extracellular polymeric substances (EPS) was analysed by biochemical and high-resolution solid state ¹³C nuclear magnetic resonance (NMR) spectroscopic methods. The EPS yield was strongly dependent on the extraction method, with the formaldehyde-NaOH method showing the best extraction efficiency. The NMR method revealed that when using the EDTA extraction method, about 40% of the EDTA accumulated in the EPS and that was responsible for the apparent high extraction yields. EPS protein content determined by the NMR method was up to 30% higher than the protein content determined using the biochemical (Lowry) method for P. putida and for A. pullulans. The average protein carbon content determined by the NMR method was approximately 70% of the total carbon content. NMR results could be supported by elemental analysis, which showed a high nitrogen content (~10%) in the EPS. The carbohydrate carbon content detected with both methods in the cell aggregates and the EPS was approximately 20% in each. In this study, quantitative ¹³C cross-polarisation magic angle spinning NMR spectroscopy was conducted on unlabeled cell strains, and EPS and could be used to quantify protein and carbohydrate of different samples. |
doi_str_mv | 10.1007/s00253-009-2218-y |
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The composition of the extracellular polymeric substances (EPS) was analysed by biochemical and high-resolution solid state ¹³C nuclear magnetic resonance (NMR) spectroscopic methods. The EPS yield was strongly dependent on the extraction method, with the formaldehyde-NaOH method showing the best extraction efficiency. The NMR method revealed that when using the EDTA extraction method, about 40% of the EDTA accumulated in the EPS and that was responsible for the apparent high extraction yields. EPS protein content determined by the NMR method was up to 30% higher than the protein content determined using the biochemical (Lowry) method for P. putida and for A. pullulans. The average protein carbon content determined by the NMR method was approximately 70% of the total carbon content. NMR results could be supported by elemental analysis, which showed a high nitrogen content (~10%) in the EPS. The carbohydrate carbon content detected with both methods in the cell aggregates and the EPS was approximately 20% in each. In this study, quantitative ¹³C cross-polarisation magic angle spinning NMR spectroscopy was conducted on unlabeled cell strains, and EPS and could be used to quantify protein and carbohydrate of different samples.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-009-2218-y</identifier><identifier>PMID: 19795119</identifier><identifier>CODEN: AMBIDG</identifier><language>eng</language><publisher>Berlin/Heidelberg: Berlin/Heidelberg : Springer-Verlag</publisher><subject>Ascomycota - metabolism ; Aureobasidium pullulans ; Bacterial Proteins - analysis ; Biofilms ; Biological and medical sciences ; Biotechnology ; Carbohydrates ; Carbon ; cell aggregates ; Cell culture ; EDTA (chelating agent) ; Extraction processes ; Formaldehyde ; Fundamental and applied biological sciences. Psychology ; Fungal Proteins - analysis ; Life Sciences ; Magnetic Resonance Spectroscopy - methods ; Methods ; Microbial Genetics and Genomics ; Microbiology ; nitrogen content ; NMR ; Nuclear magnetic resonance ; nuclear magnetic resonance spectroscopy ; Polymers ; Polymers - chemistry ; Polymers - isolation & purification ; Polymers - metabolism ; polysaccharides ; Polysaccharides - analysis ; protein content ; Proteins ; Pseudomonas putida ; Pseudomonas putida - metabolism ; Saccharides ; Sodium hydroxide ; Spectroscopy ; Spectrum analysis ; Stainless steel ; Studies</subject><ispartof>Applied microbiology and biotechnology, 2009-11, Vol.85 (1), p.197-206</ispartof><rights>Springer-Verlag 2009</rights><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c293y-b072d43395dfa1e275b90c1a8c8164ccfcd7762d0c334a27bea0b23af67c17ea3</citedby><cites>FETCH-LOGICAL-c293y-b072d43395dfa1e275b90c1a8c8164ccfcd7762d0c334a27bea0b23af67c17ea3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/229603838/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$H</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/229603838?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>314,780,784,11688,27924,27925,36060,36061,44363,74895</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=22347951$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19795119$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Metzger, Ulrich</creatorcontrib><creatorcontrib>Lankes, Ulrich</creatorcontrib><creatorcontrib>Fischpera, Kai</creatorcontrib><creatorcontrib>Frimmel, Fritz H</creatorcontrib><title>concentration of polysaccharides and proteins in EPS of Pseudomonas putida and Aureobasidum pullulans as revealed by ¹³C CPMAS NMR spectroscopy</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>Extracellular polymeric substances were extracted from the bacterial strain Pseudomonas putida and the fungal species Aureobasidium pullulans using three different methods (formaldehyde-NaOH, ethylenediaminetetraacetic acid (EDTA) and cation-exchange-resin). The composition of the extracellular polymeric substances (EPS) was analysed by biochemical and high-resolution solid state ¹³C nuclear magnetic resonance (NMR) spectroscopic methods. The EPS yield was strongly dependent on the extraction method, with the formaldehyde-NaOH method showing the best extraction efficiency. The NMR method revealed that when using the EDTA extraction method, about 40% of the EDTA accumulated in the EPS and that was responsible for the apparent high extraction yields. EPS protein content determined by the NMR method was up to 30% higher than the protein content determined using the biochemical (Lowry) method for P. putida and for A. pullulans. The average protein carbon content determined by the NMR method was approximately 70% of the total carbon content. NMR results could be supported by elemental analysis, which showed a high nitrogen content (~10%) in the EPS. The carbohydrate carbon content detected with both methods in the cell aggregates and the EPS was approximately 20% in each. In this study, quantitative ¹³C cross-polarisation magic angle spinning NMR spectroscopy was conducted on unlabeled cell strains, and EPS and could be used to quantify protein and carbohydrate of different samples.</description><subject>Ascomycota - metabolism</subject><subject>Aureobasidium pullulans</subject><subject>Bacterial Proteins - analysis</subject><subject>Biofilms</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Carbohydrates</subject><subject>Carbon</subject><subject>cell aggregates</subject><subject>Cell culture</subject><subject>EDTA (chelating agent)</subject><subject>Extraction processes</subject><subject>Formaldehyde</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fungal Proteins - analysis</subject><subject>Life Sciences</subject><subject>Magnetic Resonance Spectroscopy - methods</subject><subject>Methods</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>nitrogen content</subject><subject>NMR</subject><subject>Nuclear magnetic resonance</subject><subject>nuclear magnetic resonance spectroscopy</subject><subject>Polymers</subject><subject>Polymers - chemistry</subject><subject>Polymers - isolation & purification</subject><subject>Polymers - metabolism</subject><subject>polysaccharides</subject><subject>Polysaccharides - analysis</subject><subject>protein content</subject><subject>Proteins</subject><subject>Pseudomonas putida</subject><subject>Pseudomonas putida - metabolism</subject><subject>Saccharides</subject><subject>Sodium hydroxide</subject><subject>Spectroscopy</subject><subject>Spectrum analysis</subject><subject>Stainless steel</subject><subject>Studies</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>M0C</sourceid><recordid>eNp9kt-K1DAYxYMo7uzoA3ijQVi8qn5J2qa5HIb1D-zq4LjX4WuSrl06TTdphT6Gz-KVt_tkZpzBBUHJRSD5nZPzcULIMwavGYB8EwF4ITIAlXHOqmx-QBYsFzyDkuUPyQKYLDJZqOqEnMZ4A8B4VZaPyQlTUhWMqQX5bnxvXD8GHFvfU9_QwXdzRGO-YmitixR7S4fgR9f2kbY9Pd9s99gmusn6ne8x0mEaW4u_ydUUnK8xtnbapfOumzpMugQF981h5yytZ3r38-7Hmq43l6st_Xj5mcbBmTH4aPwwPyGPGuyie3rcl-Tq7fmX9fvs4tO7D-vVRWa4EnNWg-Q2F0IVtkHmuCxqBYZhZSpW5sY0xkpZcgtGiBy5rB1CzQU2pTRMOhRL8urgm4a7nVwc9a6NxnUpr_NT1FLkDDiHIpEv_yJv_BT6FE5zrkoQVVpLwg6QSXPE4Bo9hHaHYdYM9L4tfWhLp7b0vi09J83zo_FU75y9VxzrScDZEcBosGsC9qaNfzjORb4nE8cPXExX_bUL9wn_9_qLg6hBr_E6JOOrLQeWA6T_w1T1T0IAK1UBifgF01bCZA</recordid><startdate>200911</startdate><enddate>200911</enddate><creator>Metzger, Ulrich</creator><creator>Lankes, Ulrich</creator><creator>Fischpera, Kai</creator><creator>Frimmel, Fritz H</creator><general>Berlin/Heidelberg : Springer-Verlag</general><general>Springer Berlin Heidelberg</general><general>Springer</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>IQODW</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>7QL</scope><scope>7T7</scope><scope>7WY</scope><scope>7WZ</scope><scope>7X7</scope><scope>7XB</scope><scope>87Z</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8FL</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BEZIV</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FRNLG</scope><scope>FYUFA</scope><scope>F~G</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K60</scope><scope>K6~</scope><scope>K9.</scope><scope>L.-</scope><scope>LK8</scope><scope>M0C</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQBIZ</scope><scope>PQBZA</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope></search><sort><creationdate>200911</creationdate><title>concentration of polysaccharides and proteins in EPS of Pseudomonas putida and Aureobasidum pullulans as revealed by ¹³C CPMAS NMR spectroscopy</title><author>Metzger, Ulrich ; Lankes, Ulrich ; Fischpera, Kai ; Frimmel, Fritz H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c293y-b072d43395dfa1e275b90c1a8c8164ccfcd7762d0c334a27bea0b23af67c17ea3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Ascomycota - metabolism</topic><topic>Aureobasidium pullulans</topic><topic>Bacterial Proteins - analysis</topic><topic>Biofilms</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Carbohydrates</topic><topic>Carbon</topic><topic>cell aggregates</topic><topic>Cell culture</topic><topic>EDTA (chelating agent)</topic><topic>Extraction processes</topic><topic>Formaldehyde</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fungal Proteins - analysis</topic><topic>Life Sciences</topic><topic>Magnetic Resonance Spectroscopy - methods</topic><topic>Methods</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>nitrogen content</topic><topic>NMR</topic><topic>Nuclear magnetic resonance</topic><topic>nuclear magnetic resonance spectroscopy</topic><topic>Polymers</topic><topic>Polymers - chemistry</topic><topic>Polymers - isolation & purification</topic><topic>Polymers - metabolism</topic><topic>polysaccharides</topic><topic>Polysaccharides - analysis</topic><topic>protein content</topic><topic>Proteins</topic><topic>Pseudomonas putida</topic><topic>Pseudomonas putida - metabolism</topic><topic>Saccharides</topic><topic>Sodium hydroxide</topic><topic>Spectroscopy</topic><topic>Spectrum analysis</topic><topic>Stainless steel</topic><topic>Studies</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Metzger, Ulrich</creatorcontrib><creatorcontrib>Lankes, Ulrich</creatorcontrib><creatorcontrib>Fischpera, Kai</creatorcontrib><creatorcontrib>Frimmel, Fritz H</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>ABI/INFORM Collection</collection><collection>ABI/INFORM Global (PDF only)</collection><collection>ProQuest Health and Medical</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ABI/INFORM Global (Alumni Edition)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ABI/INFORM Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>Business Premium Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Business Premium Collection (Alumni)</collection><collection>Health Research Premium Collection</collection><collection>ABI/INFORM Global (Corporate)</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Business Collection (Alumni Edition)</collection><collection>ProQuest Business Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ABI/INFORM Professional Advanced</collection><collection>ProQuest Biological Science Collection</collection><collection>ABI/INFORM Global</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest Science Journals</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>ProQuest Biological Science Journals</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>One Business (ProQuest)</collection><collection>ProQuest One Business (Alumni)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Metzger, Ulrich</au><au>Lankes, Ulrich</au><au>Fischpera, Kai</au><au>Frimmel, Fritz H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>concentration of polysaccharides and proteins in EPS of Pseudomonas putida and Aureobasidum pullulans as revealed by ¹³C CPMAS NMR spectroscopy</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2009-11</date><risdate>2009</risdate><volume>85</volume><issue>1</issue><spage>197</spage><epage>206</epage><pages>197-206</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><coden>AMBIDG</coden><abstract>Extracellular polymeric substances were extracted from the bacterial strain Pseudomonas putida and the fungal species Aureobasidium pullulans using three different methods (formaldehyde-NaOH, ethylenediaminetetraacetic acid (EDTA) and cation-exchange-resin). The composition of the extracellular polymeric substances (EPS) was analysed by biochemical and high-resolution solid state ¹³C nuclear magnetic resonance (NMR) spectroscopic methods. The EPS yield was strongly dependent on the extraction method, with the formaldehyde-NaOH method showing the best extraction efficiency. The NMR method revealed that when using the EDTA extraction method, about 40% of the EDTA accumulated in the EPS and that was responsible for the apparent high extraction yields. EPS protein content determined by the NMR method was up to 30% higher than the protein content determined using the biochemical (Lowry) method for P. putida and for A. pullulans. The average protein carbon content determined by the NMR method was approximately 70% of the total carbon content. NMR results could be supported by elemental analysis, which showed a high nitrogen content (~10%) in the EPS. The carbohydrate carbon content detected with both methods in the cell aggregates and the EPS was approximately 20% in each. In this study, quantitative ¹³C cross-polarisation magic angle spinning NMR spectroscopy was conducted on unlabeled cell strains, and EPS and could be used to quantify protein and carbohydrate of different samples.</abstract><cop>Berlin/Heidelberg</cop><pub>Berlin/Heidelberg : Springer-Verlag</pub><pmid>19795119</pmid><doi>10.1007/s00253-009-2218-y</doi><tpages>10</tpages></addata></record> |
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subjects | Ascomycota - metabolism Aureobasidium pullulans Bacterial Proteins - analysis Biofilms Biological and medical sciences Biotechnology Carbohydrates Carbon cell aggregates Cell culture EDTA (chelating agent) Extraction processes Formaldehyde Fundamental and applied biological sciences. Psychology Fungal Proteins - analysis Life Sciences Magnetic Resonance Spectroscopy - methods Methods Microbial Genetics and Genomics Microbiology nitrogen content NMR Nuclear magnetic resonance nuclear magnetic resonance spectroscopy Polymers Polymers - chemistry Polymers - isolation & purification Polymers - metabolism polysaccharides Polysaccharides - analysis protein content Proteins Pseudomonas putida Pseudomonas putida - metabolism Saccharides Sodium hydroxide Spectroscopy Spectrum analysis Stainless steel Studies |
title | concentration of polysaccharides and proteins in EPS of Pseudomonas putida and Aureobasidum pullulans as revealed by ¹³C CPMAS NMR spectroscopy |
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