Heterologous expression of a lectin from Pleurocybella porrigens (PPL) in Phanerochaete sordida YK-624.
Identifieur interne : 000318 ( Main/Exploration ); précédent : 000317; suivant : 000319Heterologous expression of a lectin from Pleurocybella porrigens (PPL) in Phanerochaete sordida YK-624.
Auteurs : Tomohiro Suzuki [Japon] ; Hideo Dohra [Japon] ; Saori Omae [Japon] ; Yoshino Takeshima [Japon] ; Jae-Hoon Choi [Japon] ; Hirofumi Hirai [Japon] ; Hirokazu Kawagishi [Japon]Source :
- Journal of microbiological methods [ 1872-8359 ] ; 2014.
Descripteurs français
- KwdFr :
- Agaricales (génétique), Expression des gènes (MeSH), Humains (MeSH), Japon (MeSH), Lectines (biosynthèse), Lectines (génétique), Phanerochaete (génétique), Protéines recombinantes (biosynthèse), Protéines recombinantes (génétique), Régions promotrices (génétique) (MeSH), Signaux de triage des protéines (génétique), Spectrométrie de masse MALDI (MeSH), Tests d'hémagglutination (MeSH), Électrophorèse sur gel de polyacrylamide (MeSH).
- MESH :
- biosynthèse : Lectines, Protéines recombinantes.
- génétique : Agaricales, Lectines, Phanerochaete, Protéines recombinantes, Signaux de triage des protéines.
- Expression des gènes, Humains, Japon, Régions promotrices (génétique), Spectrométrie de masse MALDI, Tests d'hémagglutination, Électrophorèse sur gel de polyacrylamide.
- Wicri :
- geographic : Japon.
English descriptors
- KwdEn :
- Agaricales (genetics), Electrophoresis, Polyacrylamide Gel (MeSH), Gene Expression (MeSH), Hemagglutination Tests (MeSH), Humans (MeSH), Japan (MeSH), Lectins (biosynthesis), Lectins (genetics), Phanerochaete (genetics), Promoter Regions, Genetic (MeSH), Protein Sorting Signals (genetics), Recombinant Proteins (biosynthesis), Recombinant Proteins (genetics), Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization (MeSH).
- MESH :
- chemical , biosynthesis : Lectins, Recombinant Proteins.
- chemical , genetics : Lectins, Protein Sorting Signals, Recombinant Proteins.
- geographic : Japan.
- genetics : Agaricales, Phanerochaete.
- Electrophoresis, Polyacrylamide Gel, Gene Expression, Hemagglutination Tests, Humans, Promoter Regions, Genetic, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization.
Abstract
Pleurocybella porrigens is a mushroom-forming fungus, which had been consumed as a traditional food in Japan. However, in 2004, 55 people got poisoned by eating the mushroom and 17 people among them died of acute encephalopathy. We have already reported the purification, characterization, and cDNA cloning of a lectin from the mushroom (PPL) which might have caused the poisoning. Here, we report the heterologous expression of recombinant PPL by basidiomycete Phanerochaete sordida YK-624. The glyceraldehyde 3-phosphate dehydrogenase gene promoter was used to drive the expression of the PPL gene (ppl) in P. sordida YK-624. Furthermore, the signal peptide of lignin peroxidase which is an extracellular protein was used to secrete rPPL into extracellular region. Fifteen regenerated clones were cultured on Kirk HNHC broth, and the presence of lectin activity in the culture broth was checked by agglutination assays. The results indicated that the culture broth of rPPL-6 clone showed the strongest hemagglutination activity, and it was therefore used for subsequent analysis. The heterologous expression of rPPL by P. sordida YK-624 was confirmed by SDS-PAGE, lectin activity by the hemagglutination assay, and mass of rPPL by MALDI-TOF respectively, indicating that the extracellular secretion of rPPL as active form was successful.
DOI: 10.1016/j.mimet.2014.02.016
PubMed: 24631556
Affiliations:
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Le document en format XML
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Electronic address: ahhirai@ipc.shizuoka.ac.jp.</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529</wicri:regionArea><wicri:noRegion>Shizuoka 422-8529</wicri:noRegion></affiliation></author><author><name sortKey="Kawagishi, Hirokazu" sort="Kawagishi, Hirokazu" uniqKey="Kawagishi H" first="Hirokazu" last="Kawagishi">Hirokazu Kawagishi</name><affiliation wicri:level="1"><nlm:affiliation>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan. 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Electronic address: ahhirai@ipc.shizuoka.ac.jp.</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529</wicri:regionArea><wicri:noRegion>Shizuoka 422-8529</wicri:noRegion></affiliation></author><author><name sortKey="Kawagishi, Hirokazu" sort="Kawagishi, Hirokazu" uniqKey="Kawagishi H" first="Hirokazu" last="Kawagishi">Hirokazu Kawagishi</name><affiliation wicri:level="1"><nlm:affiliation>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan. Electronic address: achkawa@ipc.shizuoka.ac.jp.</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529</wicri:regionArea><wicri:noRegion>Shizuoka 422-8529</wicri:noRegion></affiliation></author></analytic><series><title level="j">Journal of microbiological methods</title><idno type="eISSN">1872-8359</idno><imprint><date when="2014" type="published">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agaricales (genetics)</term><term>Electrophoresis, Polyacrylamide Gel (MeSH)</term><term>Gene Expression (MeSH)</term><term>Hemagglutination Tests (MeSH)</term><term>Humans (MeSH)</term><term>Japan (MeSH)</term><term>Lectins (biosynthesis)</term><term>Lectins (genetics)</term><term>Phanerochaete (genetics)</term><term>Promoter Regions, Genetic (MeSH)</term><term>Protein Sorting Signals (genetics)</term><term>Recombinant Proteins (biosynthesis)</term><term>Recombinant Proteins (genetics)</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Agaricales (génétique)</term><term>Expression des gènes (MeSH)</term><term>Humains (MeSH)</term><term>Japon (MeSH)</term><term>Lectines (biosynthèse)</term><term>Lectines (génétique)</term><term>Phanerochaete (génétique)</term><term>Protéines recombinantes (biosynthèse)</term><term>Protéines recombinantes (génétique)</term><term>Régions promotrices (génétique) (MeSH)</term><term>Signaux de triage des protéines (génétique)</term><term>Spectrométrie de masse MALDI (MeSH)</term><term>Tests d'hémagglutination (MeSH)</term><term>Électrophorèse sur gel de polyacrylamide (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Lectins</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Lectins</term><term>Protein Sorting Signals</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="geographic" xml:lang="en"><term>Japan</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Lectines</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agaricales</term><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Agaricales</term><term>Lectines</term><term>Phanerochaete</term><term>Protéines recombinantes</term><term>Signaux de triage des protéines</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Electrophoresis, Polyacrylamide Gel</term><term>Gene Expression</term><term>Hemagglutination Tests</term><term>Humans</term><term>Promoter Regions, Genetic</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Expression des gènes</term><term>Humains</term><term>Japon</term><term>Régions promotrices (génétique)</term><term>Spectrométrie de masse MALDI</term><term>Tests d'hémagglutination</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords><keywords scheme="Wicri" type="geographic" xml:lang="fr"><term>Japon</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Pleurocybella porrigens is a mushroom-forming fungus, which had been consumed as a traditional food in Japan. However, in 2004, 55 people got poisoned by eating the mushroom and 17 people among them died of acute encephalopathy. We have already reported the purification, characterization, and cDNA cloning of a lectin from the mushroom (PPL) which might have caused the poisoning. Here, we report the heterologous expression of recombinant PPL by basidiomycete Phanerochaete sordida YK-624. The glyceraldehyde 3-phosphate dehydrogenase gene promoter was used to drive the expression of the PPL gene (ppl) in P. sordida YK-624. Furthermore, the signal peptide of lignin peroxidase which is an extracellular protein was used to secrete rPPL into extracellular region. Fifteen regenerated clones were cultured on Kirk HNHC broth, and the presence of lectin activity in the culture broth was checked by agglutination assays. The results indicated that the culture broth of rPPL-6 clone showed the strongest hemagglutination activity, and it was therefore used for subsequent analysis. The heterologous expression of rPPL by P. sordida YK-624 was confirmed by SDS-PAGE, lectin activity by the hemagglutination assay, and mass of rPPL by MALDI-TOF respectively, indicating that the extracellular secretion of rPPL as active form was successful. </div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">24631556</PMID><DateCompleted><Year>2015</Year><Month>01</Month><Day>26</Day></DateCompleted><DateRevised><Year>2014</Year><Month>04</Month><Day>18</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1872-8359</ISSN><JournalIssue CitedMedium="Internet"><Volume>100</Volume><PubDate><Year>2014</Year><Month>May</Month></PubDate></JournalIssue><Title>Journal of microbiological methods</Title><ISOAbbreviation>J Microbiol Methods</ISOAbbreviation></Journal><ArticleTitle>Heterologous expression of a lectin from Pleurocybella porrigens (PPL) in Phanerochaete sordida YK-624.</ArticleTitle><Pagination><MedlinePgn>70-6</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.mimet.2014.02.016</ELocationID><ELocationID EIdType="pii" ValidYN="Y">S0167-7012(14)00056-6</ELocationID><Abstract><AbstractText>Pleurocybella porrigens is a mushroom-forming fungus, which had been consumed as a traditional food in Japan. However, in 2004, 55 people got poisoned by eating the mushroom and 17 people among them died of acute encephalopathy. We have already reported the purification, characterization, and cDNA cloning of a lectin from the mushroom (PPL) which might have caused the poisoning. Here, we report the heterologous expression of recombinant PPL by basidiomycete Phanerochaete sordida YK-624. The glyceraldehyde 3-phosphate dehydrogenase gene promoter was used to drive the expression of the PPL gene (ppl) in P. sordida YK-624. Furthermore, the signal peptide of lignin peroxidase which is an extracellular protein was used to secrete rPPL into extracellular region. Fifteen regenerated clones were cultured on Kirk HNHC broth, and the presence of lectin activity in the culture broth was checked by agglutination assays. The results indicated that the culture broth of rPPL-6 clone showed the strongest hemagglutination activity, and it was therefore used for subsequent analysis. The heterologous expression of rPPL by P. sordida YK-624 was confirmed by SDS-PAGE, lectin activity by the hemagglutination assay, and mass of rPPL by MALDI-TOF respectively, indicating that the extracellular secretion of rPPL as active form was successful. </AbstractText><CopyrightInformation>Copyright © 2014 Elsevier B.V. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Suzuki</LastName><ForeName>Tomohiro</ForeName><Initials>T</Initials><AffiliationInfo><Affiliation>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Dohra</LastName><ForeName>Hideo</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Omae</LastName><ForeName>Saori</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Graduate School of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Takeshima</LastName><ForeName>Yoshino</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Choi</LastName><ForeName>Jae-Hoon</ForeName><Initials>JH</Initials><AffiliationInfo><Affiliation>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hirai</LastName><ForeName>Hirofumi</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan. Electronic address: ahhirai@ipc.shizuoka.ac.jp.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kawagishi</LastName><ForeName>Hirokazu</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan. Electronic address: achkawa@ipc.shizuoka.ac.jp.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2014</Year><Month>03</Month><Day>11</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>J Microbiol Methods</MedlineTA><NlmUniqueID>8306883</NlmUniqueID><ISSNLinking>0167-7012</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D037102">Lectins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D021382">Protein Sorting Signals</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000363" MajorTopicYN="N">Agaricales</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004591" MajorTopicYN="N">Electrophoresis, Polyacrylamide Gel</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015870" MajorTopicYN="Y">Gene Expression</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006386" MajorTopicYN="N">Hemagglutination Tests</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007564" MajorTopicYN="N" Type="Geographic">Japan</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D037102" MajorTopicYN="N">Lectins</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="Y">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020075" MajorTopicYN="N">Phanerochaete</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011401" MajorTopicYN="N">Promoter Regions, Genetic</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D021382" MajorTopicYN="N">Protein Sorting Signals</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D019032" MajorTopicYN="N">Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</DescriptorName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Heterologous expression</Keyword><Keyword MajorTopicYN="N">Lectin</Keyword><Keyword MajorTopicYN="N">Phanerochaete sordida YK-624</Keyword><Keyword MajorTopicYN="N">Pleurocybella porrigens</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2013</Year><Month>10</Month><Day>31</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2014</Year><Month>02</Month><Day>26</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2014</Year><Month>02</Month><Day>28</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2014</Year><Month>3</Month><Day>18</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2014</Year><Month>3</Month><Day>19</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2015</Year><Month>1</Month><Day>27</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">24631556</ArticleId><ArticleId IdType="pii">S0167-7012(14)00056-6</ArticleId><ArticleId IdType="doi">10.1016/j.mimet.2014.02.016</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Japon</li></country></list><tree><country name="Japon"><noRegion><name sortKey="Suzuki, Tomohiro" sort="Suzuki, Tomohiro" uniqKey="Suzuki T" first="Tomohiro" last="Suzuki">Tomohiro Suzuki</name></noRegion><name sortKey="Choi, Jae Hoon" sort="Choi, Jae Hoon" uniqKey="Choi J" first="Jae-Hoon" last="Choi">Jae-Hoon Choi</name><name sortKey="Dohra, Hideo" sort="Dohra, Hideo" uniqKey="Dohra H" first="Hideo" last="Dohra">Hideo Dohra</name><name sortKey="Hirai, Hirofumi" sort="Hirai, Hirofumi" uniqKey="Hirai H" first="Hirofumi" last="Hirai">Hirofumi Hirai</name><name sortKey="Kawagishi, Hirokazu" sort="Kawagishi, Hirokazu" uniqKey="Kawagishi H" first="Hirokazu" last="Kawagishi">Hirokazu Kawagishi</name><name sortKey="Omae, Saori" sort="Omae, Saori" uniqKey="Omae S" first="Saori" last="Omae">Saori Omae</name><name sortKey="Takeshima, Yoshino" sort="Takeshima, Yoshino" uniqKey="Takeshima Y" first="Yoshino" last="Takeshima">Yoshino Takeshima</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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