Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris
Chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. Whi...
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container_title | Applied microbiology and biotechnology |
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creator | He, Wenqin Zhu, Yuanyuan Shirke, Abhijeet Sun, Xiaojun Liu, Jian Gross, Richard A. Koffas, Mattheos A. G. Linhardt, Robert J. Li, Ming |
description | Chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. While the enzymes involved in the biosynthesis of chondroitin sulfates are well known, the cloning end expression of these membrane-bound Golgi enzymes continue to pose challenges. The major chondroitin-4-sulfotransferase,
Homo sapiens
C4ST-1, had been previously cloned and expressed from mammalian CHO, COS-7, and HEK 293 cells, and its activity was shown to require glycosylation. In the current study, a C4ST-1 construct was designed and expressed in both
Escherichia coli
and
Pichia pastoris
in its non-glycosylated and glycosylated forms. Both constructs showed similar activity albeit different kinetic parameters when acting on a microbially prepared unsulfated chondroitin substrate. Moreover, the glycosylated form of C4ST-1 showed lower stability than the non-glycosylated form. |
doi_str_mv | 10.1007/s00253-017-8411-5 |
format | Article |
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Homo sapiens
C4ST-1, had been previously cloned and expressed from mammalian CHO, COS-7, and HEK 293 cells, and its activity was shown to require glycosylation. In the current study, a C4ST-1 construct was designed and expressed in both
Escherichia coli
and
Pichia pastoris
in its non-glycosylated and glycosylated forms. Both constructs showed similar activity albeit different kinetic parameters when acting on a microbially prepared unsulfated chondroitin substrate. Moreover, the glycosylated form of C4ST-1 showed lower stability than the non-glycosylated form.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-017-8411-5</identifier><identifier>PMID: 28761999</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Biomedical and Life Sciences ; Biosynthesis ; Biotechnologically Relevant Enzymes and Proteins ; Biotechnology ; Chondroitin sulfate ; Cloning ; E coli ; Enzymes ; Escherichia coli ; Functional foods & nutraceuticals ; Gene expression ; Genetic aspects ; Glycosaminoglycans ; Glycosylation ; Golgi apparatus ; Life Sciences ; Microbial Genetics and Genomics ; Microbiology ; Observations ; Pichia pastoris ; Polysaccharides ; Saccharides ; Sulfates ; Sulfotransferase ; Yeast ; Yeasts (Fungi)</subject><ispartof>Applied microbiology and biotechnology, 2017-09, Vol.101 (18), p.6919-6928</ispartof><rights>Springer-Verlag GmbH Germany 2017</rights><rights>COPYRIGHT 2017 Springer</rights><rights>Applied Microbiology and Biotechnology is a copyright of Springer, 2017.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c576t-828f8edb512cdbd99c2afbcf9ea1ecd083843bd15a0fdc6a12b200b7d809593a3</citedby><cites>FETCH-LOGICAL-c576t-828f8edb512cdbd99c2afbcf9ea1ecd083843bd15a0fdc6a12b200b7d809593a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-017-8411-5$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-017-8411-5$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28761999$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>He, Wenqin</creatorcontrib><creatorcontrib>Zhu, Yuanyuan</creatorcontrib><creatorcontrib>Shirke, Abhijeet</creatorcontrib><creatorcontrib>Sun, Xiaojun</creatorcontrib><creatorcontrib>Liu, Jian</creatorcontrib><creatorcontrib>Gross, Richard A.</creatorcontrib><creatorcontrib>Koffas, Mattheos A. G.</creatorcontrib><creatorcontrib>Linhardt, Robert J.</creatorcontrib><creatorcontrib>Li, Ming</creatorcontrib><title>Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>Chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. While the enzymes involved in the biosynthesis of chondroitin sulfates are well known, the cloning end expression of these membrane-bound Golgi enzymes continue to pose challenges. The major chondroitin-4-sulfotransferase,
Homo sapiens
C4ST-1, had been previously cloned and expressed from mammalian CHO, COS-7, and HEK 293 cells, and its activity was shown to require glycosylation. In the current study, a C4ST-1 construct was designed and expressed in both
Escherichia coli
and
Pichia pastoris
in its non-glycosylated and glycosylated forms. Both constructs showed similar activity albeit different kinetic parameters when acting on a microbially prepared unsulfated chondroitin substrate. Moreover, the glycosylated form of C4ST-1 showed lower stability than the non-glycosylated form.</description><subject>Biomedical and Life Sciences</subject><subject>Biosynthesis</subject><subject>Biotechnologically Relevant Enzymes and Proteins</subject><subject>Biotechnology</subject><subject>Chondroitin sulfate</subject><subject>Cloning</subject><subject>E coli</subject><subject>Enzymes</subject><subject>Escherichia coli</subject><subject>Functional foods & nutraceuticals</subject><subject>Gene expression</subject><subject>Genetic aspects</subject><subject>Glycosaminoglycans</subject><subject>Glycosylation</subject><subject>Golgi apparatus</subject><subject>Life Sciences</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Observations</subject><subject>Pichia pastoris</subject><subject>Polysaccharides</subject><subject>Saccharides</subject><subject>Sulfates</subject><subject>Sulfotransferase</subject><subject>Yeast</subject><subject>Yeasts (Fungi)</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp1kUtr3DAUhUVoSaZpf0A2wZBNulAqyZYtLUOYtoFASl9bIesxo-CRpro2JP8-mjp9TEnRQuje7x7u0UHohJILSkj3DghhvMaEdlg0lGJ-gBa0qRkmLW1eoEVpcNxxKY7QK4A7QigTbXuIjpjoWiqlXKDvy_ttdgAhxSr5yqxTtDmFMUTc4FsM0-DTmHUE77IGV4VYLcGsXQ5mHXRl0hAqHW31aX5vNYwpB3iNXno9gHvzdB-jb--XX68-4pvbD9dXlzfY8K4dsWDCC2d7TpmxvZXSMO1746XT1BlLRC2aureUa-KtaTVlPSOk76wgksta18fofNbd5vRjcjCqTQDjhkFHlyZQVDLOhGiFKOjZP-hdmnIs2xWqJoJ1jeB_qJUenArxp3uzE1WXnFBel58lhbp4hirHuk0wKTofSn1v4O3eQGFGdz-u9ASgrr983mfpzJqcALLzapvDRucHRYna5a7m3FWJV-1yV7u1T5_MTf3G2d8Tv4IuAJsBKK24cvkv9_9VfQS5krXo</recordid><startdate>20170901</startdate><enddate>20170901</enddate><creator>He, Wenqin</creator><creator>Zhu, Yuanyuan</creator><creator>Shirke, Abhijeet</creator><creator>Sun, Xiaojun</creator><creator>Liu, Jian</creator><creator>Gross, Richard A.</creator><creator>Koffas, Mattheos A. 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G.</au><au>Linhardt, Robert J.</au><au>Li, Ming</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2017-09-01</date><risdate>2017</risdate><volume>101</volume><issue>18</issue><spage>6919</spage><epage>6928</epage><pages>6919-6928</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>Chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. While the enzymes involved in the biosynthesis of chondroitin sulfates are well known, the cloning end expression of these membrane-bound Golgi enzymes continue to pose challenges. The major chondroitin-4-sulfotransferase,
Homo sapiens
C4ST-1, had been previously cloned and expressed from mammalian CHO, COS-7, and HEK 293 cells, and its activity was shown to require glycosylation. In the current study, a C4ST-1 construct was designed and expressed in both
Escherichia coli
and
Pichia pastoris
in its non-glycosylated and glycosylated forms. Both constructs showed similar activity albeit different kinetic parameters when acting on a microbially prepared unsulfated chondroitin substrate. Moreover, the glycosylated form of C4ST-1 showed lower stability than the non-glycosylated form.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>28761999</pmid><doi>10.1007/s00253-017-8411-5</doi><tpages>10</tpages></addata></record> |
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subjects | Biomedical and Life Sciences Biosynthesis Biotechnologically Relevant Enzymes and Proteins Biotechnology Chondroitin sulfate Cloning E coli Enzymes Escherichia coli Functional foods & nutraceuticals Gene expression Genetic aspects Glycosaminoglycans Glycosylation Golgi apparatus Life Sciences Microbial Genetics and Genomics Microbiology Observations Pichia pastoris Polysaccharides Saccharides Sulfates Sulfotransferase Yeast Yeasts (Fungi) |
title | Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris |
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