Hyaluronan synthesis in cultured tobacco cells (BY-2) expressing a chlorovirus enzyme: Cytological studies
Extraction of hyaluronan from animals or microbial fermentation has risks including contamination with pathogens and microbial toxins. In this work, tobacco cultured‐cells (BY‐2) were successfully transformed with a chloroviral hyaluronan synthase (cvHAS) gene to produce hyaluronan. Cytological stud...
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| Veröffentlicht in: | Biotechnology and bioengineering 2013-04, Vol.110 (4), p.1174-1179 |
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| creator | Rakkhumkaew, Numfon Shibatani, Shigeo Kawasaki, Takeru Fujie, Makoto Yamada, Takashi |
| description | Extraction of hyaluronan from animals or microbial fermentation has risks including contamination with pathogens and microbial toxins. In this work, tobacco cultured‐cells (BY‐2) were successfully transformed with a chloroviral hyaluronan synthase (cvHAS) gene to produce hyaluronan. Cytological studies revealed accumulation of HA on the cells, and also in subcellular fractions (protoplasts, miniplasts, vacuoplasts, and vacuoles). Transgenic BY‐2 cells harboring a vSPO‐cvHAS construct containing the vacuolar targeting signal of sporamin connected to the N‐terminus of cvHAS accumulated significant amounts of HA in vacuoles. These results suggested that cvHAS successfully functions on the vacuolar membrane and synthesizes/transports HA into vacuoles. Efficient synthesis of HA using this system provides a new method for practical production of HA. Biotechnol. Bioeng. 2013; 110: 1174–1179. © 2012 Wiley Periodicals, Inc.
In this study, the authors describe the novel production of HA in cultured tobacco cells (BY‐2). The tobacco cells were successfully transformed with the chloroviral hyaluronan synthase (cv‐has) gene with or without a vacuolar targeting signal. The results showed that the sporamin vacuolar targeting signal (vSPO) operated well in BY‐2 cells, targeting the cvHAS protein to the vacuolar membrane where HA was synthesized and transported into the storage vacuole. |
| doi_str_mv | 10.1002/bit.24783 |
| format | Article |
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In this study, the authors describe the novel production of HA in cultured tobacco cells (BY‐2). The tobacco cells were successfully transformed with the chloroviral hyaluronan synthase (cv‐has) gene with or without a vacuolar targeting signal. The results showed that the sporamin vacuolar targeting signal (vSPO) operated well in BY‐2 cells, targeting the cvHAS protein to the vacuolar membrane where HA was synthesized and transported into the storage vacuole.</description><identifier>ISSN: 0006-3592</identifier><identifier>EISSN: 1097-0290</identifier><identifier>DOI: 10.1002/bit.24783</identifier><identifier>PMID: 23404209</identifier><identifier>CODEN: BIBIAU</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Base Sequence ; BY-2 ; Cell culture ; Cell Wall ; Cells, Cultured ; Chlorella virus ; Contamination ; DNA Primers ; Enzymes ; Enzymes - metabolism ; Fermentation ; Glucuronosyltransferase - genetics ; HAS ; hyaluronan ; Hyaluronan Synthases ; Hyaluronic Acid - biosynthesis ; Hyaluronic Acid - metabolism ; Nicotiana - cytology ; Nicotiana - enzymology ; Organelles - metabolism ; Pathogens ; Plants, Genetically Modified ; Polymerase Chain Reaction ; Subcellular Fractions - enzymology ; Tobacco ; Toxins</subject><ispartof>Biotechnology and bioengineering, 2013-04, Vol.110 (4), p.1174-1179</ispartof><rights>Copyright © 2012 Wiley Periodicals, Inc.</rights><rights>Copyright John Wiley and Sons, Limited Apr 2013</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5253-c50a1f7a44b12ff414cbb1288b3d964bd85380cfd491dc03894966867c306ffb3</citedby><cites>FETCH-LOGICAL-c5253-c50a1f7a44b12ff414cbb1288b3d964bd85380cfd491dc03894966867c306ffb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fbit.24783$$EPDF$$P50$$Gwiley$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fbit.24783$$EHTML$$P50$$Gwiley$$Hfree_for_read</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23404209$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rakkhumkaew, Numfon</creatorcontrib><creatorcontrib>Shibatani, Shigeo</creatorcontrib><creatorcontrib>Kawasaki, Takeru</creatorcontrib><creatorcontrib>Fujie, Makoto</creatorcontrib><creatorcontrib>Yamada, Takashi</creatorcontrib><title>Hyaluronan synthesis in cultured tobacco cells (BY-2) expressing a chlorovirus enzyme: Cytological studies</title><title>Biotechnology and bioengineering</title><addtitle>Biotechnol. Bioeng</addtitle><description>Extraction of hyaluronan from animals or microbial fermentation has risks including contamination with pathogens and microbial toxins. In this work, tobacco cultured‐cells (BY‐2) were successfully transformed with a chloroviral hyaluronan synthase (cvHAS) gene to produce hyaluronan. Cytological studies revealed accumulation of HA on the cells, and also in subcellular fractions (protoplasts, miniplasts, vacuoplasts, and vacuoles). Transgenic BY‐2 cells harboring a vSPO‐cvHAS construct containing the vacuolar targeting signal of sporamin connected to the N‐terminus of cvHAS accumulated significant amounts of HA in vacuoles. These results suggested that cvHAS successfully functions on the vacuolar membrane and synthesizes/transports HA into vacuoles. Efficient synthesis of HA using this system provides a new method for practical production of HA. Biotechnol. Bioeng. 2013; 110: 1174–1179. © 2012 Wiley Periodicals, Inc.
In this study, the authors describe the novel production of HA in cultured tobacco cells (BY‐2). The tobacco cells were successfully transformed with the chloroviral hyaluronan synthase (cv‐has) gene with or without a vacuolar targeting signal. The results showed that the sporamin vacuolar targeting signal (vSPO) operated well in BY‐2 cells, targeting the cvHAS protein to the vacuolar membrane where HA was synthesized and transported into the storage vacuole.</description><subject>Base Sequence</subject><subject>BY-2</subject><subject>Cell culture</subject><subject>Cell Wall</subject><subject>Cells, Cultured</subject><subject>Chlorella virus</subject><subject>Contamination</subject><subject>DNA Primers</subject><subject>Enzymes</subject><subject>Enzymes - metabolism</subject><subject>Fermentation</subject><subject>Glucuronosyltransferase - genetics</subject><subject>HAS</subject><subject>hyaluronan</subject><subject>Hyaluronan Synthases</subject><subject>Hyaluronic Acid - biosynthesis</subject><subject>Hyaluronic Acid - metabolism</subject><subject>Nicotiana - cytology</subject><subject>Nicotiana - enzymology</subject><subject>Organelles - metabolism</subject><subject>Pathogens</subject><subject>Plants, Genetically Modified</subject><subject>Polymerase Chain Reaction</subject><subject>Subcellular Fractions - enzymology</subject><subject>Tobacco</subject><subject>Toxins</subject><issn>0006-3592</issn><issn>1097-0290</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>EIF</sourceid><recordid>eNqFkUtPFTEUxxsjkQu68AuYJm5gMdDX9OFOLvIIRBODMa6aTqcDvfZOr-0MMnx6ChdYkBg355H8zj_nnD8A7zHawwiR_cYPe4QJSV-BGUZKVIgo9BrMEEK8orUim2Ar50VpheT8DdgklCFGkJqBxclkwphib3qYp364ctln6HtoxzCMybVwiI2xNkLrQshw5-BXRXahu1kll7PvL6GB9irEFK99GjN0_e20dJ_gfBpiiJfemgDzMLbe5bdgozMhu3ePeRv8OPpyMT-pzr8dn84_n1e2JjUtERncCcNYg0nXMcxsUyopG9oqzppW1lQi27VM4dYiKhVTnEsuLEW86xq6DXbWuqsU_4wuD3rp8_32pndxzBqzcn15F6f_RykmnONayoJ-fIEu4pj6csgDJQQTlBdqd03ZFHNOrtOr5JcmTRojfe-VLl7pB68K--FRcWyWrn0mn8wpwP4a-OuDm_6tpA9OL54kq_WEz4O7eZ4w6bfmgopa__x6rM8OyRE_-670Ib0DvUir5Q</recordid><startdate>201304</startdate><enddate>201304</enddate><creator>Rakkhumkaew, Numfon</creator><creator>Shibatani, Shigeo</creator><creator>Kawasaki, Takeru</creator><creator>Fujie, Makoto</creator><creator>Yamada, Takashi</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley Subscription Services, Inc</general><scope>BSCLL</scope><scope>24P</scope><scope>WIN</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>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7T7</scope><scope>7TA</scope><scope>7TB</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>201304</creationdate><title>Hyaluronan synthesis in cultured tobacco cells (BY-2) expressing a chlorovirus enzyme: Cytological studies</title><author>Rakkhumkaew, Numfon ; Shibatani, Shigeo ; Kawasaki, Takeru ; Fujie, Makoto ; Yamada, Takashi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5253-c50a1f7a44b12ff414cbb1288b3d964bd85380cfd491dc03894966867c306ffb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Base Sequence</topic><topic>BY-2</topic><topic>Cell culture</topic><topic>Cell Wall</topic><topic>Cells, Cultured</topic><topic>Chlorella virus</topic><topic>Contamination</topic><topic>DNA Primers</topic><topic>Enzymes</topic><topic>Enzymes - metabolism</topic><topic>Fermentation</topic><topic>Glucuronosyltransferase - genetics</topic><topic>HAS</topic><topic>hyaluronan</topic><topic>Hyaluronan Synthases</topic><topic>Hyaluronic Acid - biosynthesis</topic><topic>Hyaluronic Acid - metabolism</topic><topic>Nicotiana - cytology</topic><topic>Nicotiana - enzymology</topic><topic>Organelles - metabolism</topic><topic>Pathogens</topic><topic>Plants, Genetically Modified</topic><topic>Polymerase Chain Reaction</topic><topic>Subcellular Fractions - enzymology</topic><topic>Tobacco</topic><topic>Toxins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rakkhumkaew, Numfon</creatorcontrib><creatorcontrib>Shibatani, Shigeo</creatorcontrib><creatorcontrib>Kawasaki, Takeru</creatorcontrib><creatorcontrib>Fujie, Makoto</creatorcontrib><creatorcontrib>Yamada, Takashi</creatorcontrib><collection>Istex</collection><collection>Wiley Online Library (Open Access Collection)</collection><collection>Wiley Online Library (Open Access Collection)</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biotechnology and bioengineering</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rakkhumkaew, Numfon</au><au>Shibatani, Shigeo</au><au>Kawasaki, Takeru</au><au>Fujie, Makoto</au><au>Yamada, Takashi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Hyaluronan synthesis in cultured tobacco cells (BY-2) expressing a chlorovirus enzyme: Cytological studies</atitle><jtitle>Biotechnology and bioengineering</jtitle><addtitle>Biotechnol. Bioeng</addtitle><date>2013-04</date><risdate>2013</risdate><volume>110</volume><issue>4</issue><spage>1174</spage><epage>1179</epage><pages>1174-1179</pages><issn>0006-3592</issn><eissn>1097-0290</eissn><coden>BIBIAU</coden><abstract>Extraction of hyaluronan from animals or microbial fermentation has risks including contamination with pathogens and microbial toxins. In this work, tobacco cultured‐cells (BY‐2) were successfully transformed with a chloroviral hyaluronan synthase (cvHAS) gene to produce hyaluronan. Cytological studies revealed accumulation of HA on the cells, and also in subcellular fractions (protoplasts, miniplasts, vacuoplasts, and vacuoles). Transgenic BY‐2 cells harboring a vSPO‐cvHAS construct containing the vacuolar targeting signal of sporamin connected to the N‐terminus of cvHAS accumulated significant amounts of HA in vacuoles. These results suggested that cvHAS successfully functions on the vacuolar membrane and synthesizes/transports HA into vacuoles. Efficient synthesis of HA using this system provides a new method for practical production of HA. Biotechnol. Bioeng. 2013; 110: 1174–1179. © 2012 Wiley Periodicals, Inc.
In this study, the authors describe the novel production of HA in cultured tobacco cells (BY‐2). The tobacco cells were successfully transformed with the chloroviral hyaluronan synthase (cv‐has) gene with or without a vacuolar targeting signal. The results showed that the sporamin vacuolar targeting signal (vSPO) operated well in BY‐2 cells, targeting the cvHAS protein to the vacuolar membrane where HA was synthesized and transported into the storage vacuole.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>23404209</pmid><doi>10.1002/bit.24783</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Base Sequence BY-2 Cell culture Cell Wall Cells, Cultured Chlorella virus Contamination DNA Primers Enzymes Enzymes - metabolism Fermentation Glucuronosyltransferase - genetics HAS hyaluronan Hyaluronan Synthases Hyaluronic Acid - biosynthesis Hyaluronic Acid - metabolism Nicotiana - cytology Nicotiana - enzymology Organelles - metabolism Pathogens Plants, Genetically Modified Polymerase Chain Reaction Subcellular Fractions - enzymology Tobacco Toxins |
| title | Hyaluronan synthesis in cultured tobacco cells (BY-2) expressing a chlorovirus enzyme: Cytological studies |
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