Mixed‐Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase
The mixed‐linkage (1→3),(1→4)‐d‐glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. It holds potential for application in the brewery, animal feed, and biofuel industries. Several defined MLG oligosaccharides obtained by au...
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| Veröffentlicht in: | Chemistry : a European journal 2017-03, Vol.23 (13), p.3191-3196 |
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| creator | Dallabernardina, Pietro Schuhmacher, Frank Seeberger, Peter H. Pfrengle, Fabian |
| description | The mixed‐linkage (1→3),(1→4)‐d‐glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. It holds potential for application in the brewery, animal feed, and biofuel industries. Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C‐3 or C‐4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. Light‐induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. After incubation of the MLG oligosaccharides with lichenase, the digestion products were analyzed by HPLC‐MS. These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed‐linkage β‐glucanases.
Assembly and deconstruction: The automated glycan assembly of mixed‐linkage glucan oligosaccharides as biochemical tools for analyzing the substrate specificities of lichenase and other mixed‐linkage β‐glucanases is reported (see scheme). |
| doi_str_mv | 10.1002/chem.201605479 |
| format | Article |
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Assembly and deconstruction: The automated glycan assembly of mixed‐linkage glucan oligosaccharides as biochemical tools for analyzing the substrate specificities of lichenase and other mixed‐linkage β‐glucanases is reported (see scheme).</description><identifier>ISSN: 0947-6539</identifier><identifier>EISSN: 1521-3765</identifier><identifier>DOI: 10.1002/chem.201605479</identifier><identifier>PMID: 28084659</identifier><identifier>CODEN: CEUJED</identifier><language>eng</language><publisher>Germany: Wiley Subscription Services, Inc</publisher><subject>Assembly ; automated glycan assembly ; Automation ; Bacillus subtilis ; Bacillus subtilis - chemistry ; Bacillus subtilis - enzymology ; Bacillus subtilis - metabolism ; Biochemistry ; carbohydrates ; Catalytic Domain ; Chemistry ; Chromatography, High Pressure Liquid ; Digestion ; Glucan ; Glucans - chemistry ; Glucans - metabolism ; Glycan ; Glycoside Hydrolases - chemistry ; Glycoside Hydrolases - metabolism ; lichenase ; Mass Spectrometry ; mixed-linkage glucan ; Oligosaccharides ; Oligosaccharides - chemistry ; Oligosaccharides - metabolism ; plant cell wall ; Substrate Specificity ; Substrates</subject><ispartof>Chemistry : a European journal, 2017-03, Vol.23 (13), p.3191-3196</ispartof><rights>2017 Wiley‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><rights>2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.</rights><rights>2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5169-b742816901b502aefd9966eb9276b4d21cfd8009de9d0ee1d34f0296bbb5604d3</citedby><cites>FETCH-LOGICAL-c5169-b742816901b502aefd9966eb9276b4d21cfd8009de9d0ee1d34f0296bbb5604d3</cites><orcidid>0000-0002-2586-4118 ; 0000-0003-2206-6636</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fchem.201605479$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fchem.201605479$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28084659$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Dallabernardina, Pietro</creatorcontrib><creatorcontrib>Schuhmacher, Frank</creatorcontrib><creatorcontrib>Seeberger, Peter H.</creatorcontrib><creatorcontrib>Pfrengle, Fabian</creatorcontrib><title>Mixed‐Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase</title><title>Chemistry : a European journal</title><addtitle>Chemistry</addtitle><description>The mixed‐linkage (1→3),(1→4)‐d‐glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. It holds potential for application in the brewery, animal feed, and biofuel industries. Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C‐3 or C‐4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. Light‐induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. After incubation of the MLG oligosaccharides with lichenase, the digestion products were analyzed by HPLC‐MS. These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed‐linkage β‐glucanases.
Assembly and deconstruction: The automated glycan assembly of mixed‐linkage glucan oligosaccharides as biochemical tools for analyzing the substrate specificities of lichenase and other mixed‐linkage β‐glucanases is reported (see scheme).</description><subject>Assembly</subject><subject>automated glycan assembly</subject><subject>Automation</subject><subject>Bacillus subtilis</subject><subject>Bacillus subtilis - chemistry</subject><subject>Bacillus subtilis - enzymology</subject><subject>Bacillus subtilis - metabolism</subject><subject>Biochemistry</subject><subject>carbohydrates</subject><subject>Catalytic Domain</subject><subject>Chemistry</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Digestion</subject><subject>Glucan</subject><subject>Glucans - chemistry</subject><subject>Glucans - metabolism</subject><subject>Glycan</subject><subject>Glycoside Hydrolases - chemistry</subject><subject>Glycoside Hydrolases - metabolism</subject><subject>lichenase</subject><subject>Mass Spectrometry</subject><subject>mixed-linkage glucan</subject><subject>Oligosaccharides</subject><subject>Oligosaccharides - chemistry</subject><subject>Oligosaccharides - metabolism</subject><subject>plant cell wall</subject><subject>Substrate Specificity</subject><subject>Substrates</subject><issn>0947-6539</issn><issn>1521-3765</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqN0stu00AUBmALgWgobFmikdiwSZi7PcsoLSlSqiKlrK25HDdTbE-YsQHveIQ-I0_CRClFYkM3c5G-80tH5xTFa4IXBGP63u6gW1BMJBa8VE-KGRGUzFkpxdNihhUv51IwdVK8SOkWY6wkY8-LE1rhikuhZsXdpf8B7tfPu43vv-gbQOt2tLpHV62_CUlbu9PRO0joUwxutOCQmdByHEKnh_xZt9NBL1OCzrQT2kL8BkgndB1CezjRGQwQO98DGnaAtqNJQ8ylaLsH6xtv_TCh0KCNz530OsHL4lmj2wSv7u_T4vOH8-vVxXxztf64Wm7mVhCp5qbktMoPTIzAVEPjlJISjKKlNNxRYhtX5X4dKIcBiGO8wVRJY4yQmDt2Wrw75u5j-DpCGurOJwttq3sIY6pJpVillGL0EbSqVCkEZY-gknCevcj07T_0Noyxzz1nVXJGiOJVVoujsjGkFKGp99F3Ok41wfVhA-rDBtQPG5AL3tzHjqYD98D_jDwDdQTffQvTf-Lq1cX55d_w3wrhvk0</recordid><startdate>20170302</startdate><enddate>20170302</enddate><creator>Dallabernardina, Pietro</creator><creator>Schuhmacher, Frank</creator><creator>Seeberger, Peter H.</creator><creator>Pfrengle, Fabian</creator><general>Wiley Subscription Services, 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>7SR</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>K9.</scope><scope>7X8</scope><scope>7QO</scope><scope>FR3</scope><scope>P64</scope><orcidid>https://orcid.org/0000-0002-2586-4118</orcidid><orcidid>https://orcid.org/0000-0003-2206-6636</orcidid></search><sort><creationdate>20170302</creationdate><title>Mixed‐Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase</title><author>Dallabernardina, Pietro ; Schuhmacher, Frank ; Seeberger, Peter H. ; Pfrengle, Fabian</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5169-b742816901b502aefd9966eb9276b4d21cfd8009de9d0ee1d34f0296bbb5604d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Assembly</topic><topic>automated glycan assembly</topic><topic>Automation</topic><topic>Bacillus subtilis</topic><topic>Bacillus subtilis - chemistry</topic><topic>Bacillus subtilis - enzymology</topic><topic>Bacillus subtilis - metabolism</topic><topic>Biochemistry</topic><topic>carbohydrates</topic><topic>Catalytic Domain</topic><topic>Chemistry</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Digestion</topic><topic>Glucan</topic><topic>Glucans - chemistry</topic><topic>Glucans - metabolism</topic><topic>Glycan</topic><topic>Glycoside Hydrolases - chemistry</topic><topic>Glycoside Hydrolases - metabolism</topic><topic>lichenase</topic><topic>Mass Spectrometry</topic><topic>mixed-linkage glucan</topic><topic>Oligosaccharides</topic><topic>Oligosaccharides - chemistry</topic><topic>Oligosaccharides - metabolism</topic><topic>plant cell wall</topic><topic>Substrate Specificity</topic><topic>Substrates</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Dallabernardina, Pietro</creatorcontrib><creatorcontrib>Schuhmacher, Frank</creatorcontrib><creatorcontrib>Seeberger, Peter H.</creatorcontrib><creatorcontrib>Pfrengle, Fabian</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Engineered Materials Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Chemistry : a European journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Dallabernardina, Pietro</au><au>Schuhmacher, Frank</au><au>Seeberger, Peter H.</au><au>Pfrengle, Fabian</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Mixed‐Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase</atitle><jtitle>Chemistry : a European journal</jtitle><addtitle>Chemistry</addtitle><date>2017-03-02</date><risdate>2017</risdate><volume>23</volume><issue>13</issue><spage>3191</spage><epage>3196</epage><pages>3191-3196</pages><issn>0947-6539</issn><eissn>1521-3765</eissn><coden>CEUJED</coden><abstract>The mixed‐linkage (1→3),(1→4)‐d‐glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. It holds potential for application in the brewery, animal feed, and biofuel industries. Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C‐3 or C‐4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. Light‐induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. After incubation of the MLG oligosaccharides with lichenase, the digestion products were analyzed by HPLC‐MS. These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed‐linkage β‐glucanases.
Assembly and deconstruction: The automated glycan assembly of mixed‐linkage glucan oligosaccharides as biochemical tools for analyzing the substrate specificities of lichenase and other mixed‐linkage β‐glucanases is reported (see scheme).</abstract><cop>Germany</cop><pub>Wiley Subscription Services, Inc</pub><pmid>28084659</pmid><doi>10.1002/chem.201605479</doi><tpages>6</tpages><orcidid>https://orcid.org/0000-0002-2586-4118</orcidid><orcidid>https://orcid.org/0000-0003-2206-6636</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Assembly automated glycan assembly Automation Bacillus subtilis Bacillus subtilis - chemistry Bacillus subtilis - enzymology Bacillus subtilis - metabolism Biochemistry carbohydrates Catalytic Domain Chemistry Chromatography, High Pressure Liquid Digestion Glucan Glucans - chemistry Glucans - metabolism Glycan Glycoside Hydrolases - chemistry Glycoside Hydrolases - metabolism lichenase Mass Spectrometry mixed-linkage glucan Oligosaccharides Oligosaccharides - chemistry Oligosaccharides - metabolism plant cell wall Substrate Specificity Substrates |
| title | Mixed‐Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase |
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