Tandem repeated expression of lunasin gene in Pichia pastoris and its anti-inflammatory activity in vitro.
Identifieur interne : 000471 ( Main/Exploration ); précédent : 000470; suivant : 000472Tandem repeated expression of lunasin gene in Pichia pastoris and its anti-inflammatory activity in vitro.
Auteurs : Yingying Zhu [Belgique] ; Everaert Nadia [Belgique] ; Yang Yao [République populaire de Chine] ; Zhenxing Shi [Belgique] ; Guixing Ren [République populaire de Chine]Source :
- Journal of bioscience and bioengineering [ 1347-4421 ] ; 2018.
Descripteurs français
- KwdFr :
- Animaux (MeSH), Anti-inflammatoires (métabolisme), Cellules cultivées (MeSH), Clonage moléculaire (méthodes), Lipopolysaccharides (MeSH), Macrophages (effets des médicaments et des substances chimiques), Macrophages (métabolisme), Multimérisation de protéines (génétique), Organismes génétiquement modifiés (MeSH), Pichia (génétique), Pichia (métabolisme), Protéines de soja (génétique), Protéines de soja (métabolisme), Protéines recombinantes (génétique), Protéines recombinantes (métabolisme), Régulation de l'expression des gènes fongiques (MeSH), Souris (MeSH), Séquences répétées en tandem (génétique).
- MESH :
- effets des médicaments et des substances chimiques : Macrophages.
- génétique : Multimérisation de protéines, Pichia, Protéines de soja, Protéines recombinantes, Séquences répétées en tandem.
- métabolisme : Anti-inflammatoires, Macrophages, Pichia, Protéines de soja, Protéines recombinantes.
- méthodes : Clonage moléculaire.
- Animaux, Cellules cultivées, Lipopolysaccharides, Organismes génétiquement modifiés, Régulation de l'expression des gènes fongiques, Souris.
English descriptors
- KwdEn :
- Animals (MeSH), Anti-Inflammatory Agents (metabolism), Cells, Cultured (MeSH), Cloning, Molecular (methods), Gene Expression Regulation, Fungal (MeSH), Lipopolysaccharides (MeSH), Macrophages (drug effects), Macrophages (metabolism), Mice (MeSH), Organisms, Genetically Modified (MeSH), Pichia (genetics), Pichia (metabolism), Protein Multimerization (genetics), Recombinant Proteins (genetics), Recombinant Proteins (metabolism), Soybean Proteins (genetics), Soybean Proteins (metabolism), Tandem Repeat Sequences (genetics).
- MESH :
- chemical , genetics : Recombinant Proteins, Soybean Proteins.
- chemical , metabolism : Anti-Inflammatory Agents, Recombinant Proteins, Soybean Proteins.
- drug effects : Macrophages.
- genetics : Pichia, Protein Multimerization, Tandem Repeat Sequences.
- metabolism : Macrophages, Pichia.
- methods : Cloning, Molecular.
- Animals, Cells, Cultured, Gene Expression Regulation, Fungal, Lipopolysaccharides, Mice, Organisms, Genetically Modified.
Abstract
Lunasin is a novel promising health-beneficial peptide derived from soybean. However, the application of lunasin is limited by its high cost. In this study, we developed a successful protocol for expression of a dimer formation protein containing 4 tandem repeated lunasin analogs (lunasin-4) in Pichia pastoris. The expression level at the optimal condition (initial pH 7.0, 1.0% final methanol concentration and induction for 72 h at 26 °C) was 0.24 mg/mL cell-free broth. Lunasin analog, obtained from purified lunasin-4 protein through enterokinase digestion and ultrafiltration, significantly decreased (p < 0.05) the release of nitric oxide (NO), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in a dose-dependent manner. In addition, intracellular signaling array analysis demonstrated down-regulated levels of phosphorylated Akt, mechanistic target of rapamycin (mTOR) and p70 s6 kinase (p70s6k) and an up-regulated level of glycogen synthase kinase-3β (GSK-3β) after lunasin analog treatment. These results suggest that lunasin analog exerted anti-inflammatory activities in LPS-stimulated RAW264.7 cells partly via inhibiting the activation of Akt/mTOR/p70s6k signaling pathway. In conclusion, this study provides a potential strategy for recombinant production of bioactive lunasin in industry.
DOI: 10.1016/j.jbiosc.2018.01.013
PubMed: 29506820
Affiliations:
- Belgique, République populaire de Chine
- Province de Liège, Région wallonne
- Liège, Pékin
- Université de Liège
Links toward previous steps (curation, corpus...)
Le document en format XML
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Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, TERRA Teaching and Research Centre, University of Liège, Passage des Déportés 2, 5030 Gembloux, Belgium.</nlm:affiliation><country xml:lang="fr">Belgique</country><wicri:regionArea>Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), 80 South Xueyuan Road, Haidian, Beijing 100081, People's Republic of China; Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, TERRA Teaching and Research Centre, University of Liège, Passage des Déportés 2, 5030 Gembloux</wicri:regionArea><orgName type="university">Université de Liège</orgName><placeName><settlement type="city">Liège</settlement><region type="region" nuts="1">Région wallonne</region><region type="province" nuts="1">Province de Liège</region></placeName></affiliation></author><author><name sortKey="Nadia, Everaert" sort="Nadia, Everaert" uniqKey="Nadia E" first="Everaert" last="Nadia">Everaert Nadia</name><affiliation wicri:level="4"><nlm:affiliation>Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, TERRA Teaching and Research Centre, University of Liège, Passage des Déportés 2, 5030 Gembloux, Belgium.</nlm:affiliation><country xml:lang="fr">Belgique</country><wicri:regionArea>Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, TERRA Teaching and Research Centre, University of Liège, Passage des Déportés 2, 5030 Gembloux</wicri:regionArea><orgName type="university">Université de Liège</orgName><placeName><settlement type="city">Liège</settlement><region type="region" nuts="1">Région wallonne</region><region type="province" nuts="1">Province de Liège</region></placeName></affiliation></author><author><name sortKey="Yao, Yang" sort="Yao, Yang" uniqKey="Yao Y" first="Yang" last="Yao">Yang Yao</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), 80 South Xueyuan Road, Haidian, Beijing 100081, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), 80 South Xueyuan Road, Haidian, Beijing 100081</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Shi, Zhenxing" sort="Shi, Zhenxing" uniqKey="Shi Z" first="Zhenxing" last="Shi">Zhenxing Shi</name><affiliation wicri:level="4"><nlm:affiliation>Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), 80 South Xueyuan Road, Haidian, Beijing 100081, People's Republic of China; 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However, the application of lunasin is limited by its high cost. In this study, we developed a successful protocol for expression of a dimer formation protein containing 4 tandem repeated lunasin analogs (lunasin-4) in Pichia pastoris. The expression level at the optimal condition (initial pH 7.0, 1.0% final methanol concentration and induction for 72 h at 26 °C) was 0.24 mg/mL cell-free broth. Lunasin analog, obtained from purified lunasin-4 protein through enterokinase digestion and ultrafiltration, significantly decreased (p < 0.05) the release of nitric oxide (NO), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in a dose-dependent manner. In addition, intracellular signaling array analysis demonstrated down-regulated levels of phosphorylated Akt, mechanistic target of rapamycin (mTOR) and p70 s6 kinase (p70s6k) and an up-regulated level of glycogen synthase kinase-3β (GSK-3β) after lunasin analog treatment. These results suggest that lunasin analog exerted anti-inflammatory activities in LPS-stimulated RAW264.7 cells partly via inhibiting the activation of Akt/mTOR/p70s6k signaling pathway. In conclusion, this study provides a potential strategy for recombinant production of bioactive lunasin in industry.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" IndexingMethod="Curated" Owner="NLM"><PMID Version="1">29506820</PMID><DateCompleted><Year>2018</Year><Month>08</Month><Day>31</Day></DateCompleted><DateRevised><Year>2018</Year><Month>12</Month><Day>02</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1347-4421</ISSN><JournalIssue CitedMedium="Internet"><Volume>126</Volume><Issue>1</Issue><PubDate><Year>2018</Year><Month>Jul</Month></PubDate></JournalIssue><Title>Journal of bioscience and bioengineering</Title><ISOAbbreviation>J Biosci Bioeng</ISOAbbreviation></Journal><ArticleTitle>Tandem repeated expression of lunasin gene in Pichia pastoris and its anti-inflammatory activity in vitro.</ArticleTitle><Pagination><MedlinePgn>1-8</MedlinePgn></Pagination><ELocationID EIdType="pii" ValidYN="Y">S1389-1723(17)31218-5</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.jbiosc.2018.01.013</ELocationID><Abstract><AbstractText>Lunasin is a novel promising health-beneficial peptide derived from soybean. However, the application of lunasin is limited by its high cost. In this study, we developed a successful protocol for expression of a dimer formation protein containing 4 tandem repeated lunasin analogs (lunasin-4) in Pichia pastoris. The expression level at the optimal condition (initial pH 7.0, 1.0% final methanol concentration and induction for 72 h at 26 °C) was 0.24 mg/mL cell-free broth. Lunasin analog, obtained from purified lunasin-4 protein through enterokinase digestion and ultrafiltration, significantly decreased (p < 0.05) the release of nitric oxide (NO), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in a dose-dependent manner. In addition, intracellular signaling array analysis demonstrated down-regulated levels of phosphorylated Akt, mechanistic target of rapamycin (mTOR) and p70 s6 kinase (p70s6k) and an up-regulated level of glycogen synthase kinase-3β (GSK-3β) after lunasin analog treatment. These results suggest that lunasin analog exerted anti-inflammatory activities in LPS-stimulated RAW264.7 cells partly via inhibiting the activation of Akt/mTOR/p70s6k signaling pathway. In conclusion, this study provides a potential strategy for recombinant production of bioactive lunasin in industry.</AbstractText><CopyrightInformation>Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Zhu</LastName><ForeName>Yingying</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), 80 South Xueyuan Road, Haidian, Beijing 100081, People's Republic of China; Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, TERRA Teaching and Research Centre, University of Liège, Passage des Déportés 2, 5030 Gembloux, Belgium.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Nadia</LastName><ForeName>Everaert</ForeName><Initials>E</Initials><AffiliationInfo><Affiliation>Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, TERRA Teaching and Research Centre, University of Liège, Passage des Déportés 2, 5030 Gembloux, Belgium.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yao</LastName><ForeName>Yang</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), 80 South Xueyuan Road, Haidian, Beijing 100081, People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Shi</LastName><ForeName>Zhenxing</ForeName><Initials>Z</Initials><AffiliationInfo><Affiliation>Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), 80 South Xueyuan Road, Haidian, Beijing 100081, People's Republic of China; Laboratory of Biomass and Green Technologies, Gembloux Agro-Bio Tech, University of Liege, Passage des Déportés 2, 5030 Gembloux, Belgium.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ren</LastName><ForeName>Guixing</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), 80 South Xueyuan Road, Haidian, Beijing 100081, People's Republic of China. Electronic address: renguixing@caas.cn.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2018</Year><Month>03</Month><Day>03</Day></ArticleDate></Article><MedlineJournalInfo><Country>Japan</Country><MedlineTA>J Biosci Bioeng</MedlineTA><NlmUniqueID>100888800</NlmUniqueID><ISSNLinking>1347-4421</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000893">Anti-Inflammatory Agents</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C120514">GM2S-1 protein, Glycine max</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D008070">Lipopolysaccharides</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant 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MajorTopicYN="N">Lipopolysaccharides</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008264" MajorTopicYN="N">Macrophages</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D051379" MajorTopicYN="N">Mice</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D030781" MajorTopicYN="N">Organisms, Genetically Modified</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010843" MajorTopicYN="N">Pichia</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D055503" MajorTopicYN="N">Protein Multimerization</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D030262" MajorTopicYN="N">Soybean Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020080" MajorTopicYN="N">Tandem Repeat Sequences</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Akt/mTOR/p70s6k pathway</Keyword><Keyword MajorTopicYN="N">Anti-inflammatory</Keyword><Keyword MajorTopicYN="N">Fermentation conditions</Keyword><Keyword MajorTopicYN="N">Lunasin</Keyword><Keyword MajorTopicYN="N">Pichia pastoris</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2017</Year><Month>12</Month><Day>15</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2018</Year><Month>01</Month><Day>16</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2018</Year><Month>01</Month><Day>18</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2018</Year><Month>3</Month><Day>7</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2018</Year><Month>9</Month><Day>1</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2018</Year><Month>3</Month><Day>7</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">29506820</ArticleId><ArticleId IdType="pii">S1389-1723(17)31218-5</ArticleId><ArticleId IdType="doi">10.1016/j.jbiosc.2018.01.013</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Belgique</li><li>République populaire de Chine</li></country><region><li>Province de Liège</li><li>Région wallonne</li></region><settlement><li>Liège</li><li>Pékin</li></settlement><orgName><li>Université de Liège</li></orgName></list><tree><country name="Belgique"><region name="Région wallonne"><name sortKey="Zhu, Yingying" sort="Zhu, Yingying" uniqKey="Zhu Y" first="Yingying" last="Zhu">Yingying Zhu</name></region><name sortKey="Nadia, Everaert" sort="Nadia, Everaert" uniqKey="Nadia E" first="Everaert" last="Nadia">Everaert Nadia</name><name sortKey="Shi, Zhenxing" sort="Shi, Zhenxing" uniqKey="Shi Z" first="Zhenxing" last="Shi">Zhenxing Shi</name></country><country name="République populaire de Chine"><noRegion><name sortKey="Yao, Yang" sort="Yao, Yang" uniqKey="Yao Y" first="Yang" last="Yao">Yang Yao</name></noRegion><name sortKey="Ren, Guixing" sort="Ren, Guixing" uniqKey="Ren G" first="Guixing" last="Ren">Guixing Ren</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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