An improved process for the production of highly purified recombinant thaumatin tagged-variants.
Identifieur interne : 000080 ( Main/Curation ); précédent : 000079; suivant : 000081An improved process for the production of highly purified recombinant thaumatin tagged-variants.
Auteurs : Robert D. Healey [Australie] ; Helene Lebhar [Australie] ; Simon Hornung [Australie] ; Pall Thordarson [Australie] ; Christopher P. Marquis [Australie]Source :
- Food chemistry [ 1873-7072 ] ; 2017.
Descripteurs français
- KwdFr :
- MESH :
- composition chimique : Protéines végétales.
- Marantaceae, Pichia, Spectrométrie de masse en tandem, Édulcorants.
English descriptors
- KwdEn :
- MESH :
- chemical , chemistry : Plant Proteins.
- Marantaceae, Pichia, Sweetening Agents, Tandem Mass Spectrometry.
Abstract
The sweetest tasting molecule known is the protein thaumatin, first isolated from the katemfe fruit, Thaumatococcus daniellii. Thaumatin is used in the food and beverage industry as a low-calorie sugar substitute. Thaumatin interacts with taste receptors in the oral cavity eliciting a persistent sweet taste and a bitter, liquorice flavor. Recombinant thaumatin was expressed in Pichia pastoris and through a co-expression strategy with a molecular chaperone, yields of one engineered thaumatin variant increased by greater than two-fold. A detailed purification strategy for thaumatin is reported resulting in a homogenous sample recovered at a yield of 42%. The recombinant thaumatins were extensively characterised using size exclusion chromatography for homogeneity, reversed-phase HPLC for purity (99%), peptide digest LC-MS/MS for sequence determination, and circular dichroism and tryptophan fluorescence spectroscopies for conformational characterisation. These new thaumatin variants are amenable for bioconjugation, providing chemical biology tools for thaumatin:taste receptor interaction studies.
DOI: 10.1016/j.foodchem.2017.06.018
PubMed: 28764073
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Healey</name><affiliation wicri:level="1"><nlm:affiliation>School of Chemistry, The Australian Centre for Nanomedicine and the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney, NSW, Australia. Electronic address: robert.healey@unsw.edu.au.</nlm:affiliation><country xml:lang="fr">Australie</country><wicri:regionArea>School of Chemistry, The Australian Centre for Nanomedicine and the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney, NSW</wicri:regionArea></affiliation></author><author><name sortKey="Lebhar, Helene" sort="Lebhar, Helene" uniqKey="Lebhar H" first="Helene" last="Lebhar">Helene Lebhar</name><affiliation wicri:level="1"><nlm:affiliation>School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia. Electronic address: h.lebhar@unsw.edu.au.</nlm:affiliation><country xml:lang="fr">Australie</country><wicri:regionArea>School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW</wicri:regionArea></affiliation></author><author><name sortKey="Hornung, Simon" sort="Hornung, Simon" uniqKey="Hornung S" first="Simon" last="Hornung">Simon Hornung</name><affiliation wicri:level="1"><nlm:affiliation>School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia. Electronic address: si.hornung@web.de.</nlm:affiliation><country xml:lang="fr">Australie</country><wicri:regionArea>School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW</wicri:regionArea></affiliation></author><author><name sortKey="Thordarson, Pall" sort="Thordarson, Pall" uniqKey="Thordarson P" first="Pall" last="Thordarson">Pall Thordarson</name><affiliation wicri:level="1"><nlm:affiliation>School of Chemistry, The Australian Centre for Nanomedicine and the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney, NSW, Australia. Electronic address: p.thordarson@unsw.edu.au.</nlm:affiliation><country xml:lang="fr">Australie</country><wicri:regionArea>School of Chemistry, The Australian Centre for Nanomedicine and the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney, NSW</wicri:regionArea></affiliation></author><author><name sortKey="Marquis, Christopher P" sort="Marquis, Christopher P" uniqKey="Marquis C" first="Christopher P" last="Marquis">Christopher P. Marquis</name><affiliation wicri:level="1"><nlm:affiliation>School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia. Electronic address: c.marquis@unsw.edu.au.</nlm:affiliation><country xml:lang="fr">Australie</country><wicri:regionArea>School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW</wicri:regionArea></affiliation></author></analytic><series><title level="j">Food chemistry</title><idno type="eISSN">1873-7072</idno><imprint><date when="2017" type="published">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Marantaceae (MeSH)</term><term>Pichia (MeSH)</term><term>Plant Proteins (chemistry)</term><term>Sweetening Agents (MeSH)</term><term>Tandem Mass Spectrometry (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Marantaceae (MeSH)</term><term>Pichia (MeSH)</term><term>Protéines végétales (composition chimique)</term><term>Spectrométrie de masse en tandem (MeSH)</term><term>Édulcorants (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Plant Proteins</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Protéines végétales</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Marantaceae</term><term>Pichia</term><term>Sweetening Agents</term><term>Tandem Mass Spectrometry</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Marantaceae</term><term>Pichia</term><term>Spectrométrie de masse en tandem</term><term>Édulcorants</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The sweetest tasting molecule known is the protein thaumatin, first isolated from the katemfe fruit, Thaumatococcus daniellii. Thaumatin is used in the food and beverage industry as a low-calorie sugar substitute. Thaumatin interacts with taste receptors in the oral cavity eliciting a persistent sweet taste and a bitter, liquorice flavor. Recombinant thaumatin was expressed in Pichia pastoris and through a co-expression strategy with a molecular chaperone, yields of one engineered thaumatin variant increased by greater than two-fold. A detailed purification strategy for thaumatin is reported resulting in a homogenous sample recovered at a yield of 42%. The recombinant thaumatins were extensively characterised using size exclusion chromatography for homogeneity, reversed-phase HPLC for purity (99%), peptide digest LC-MS/MS for sequence determination, and circular dichroism and tryptophan fluorescence spectroscopies for conformational characterisation. These new thaumatin variants are amenable for bioconjugation, providing chemical biology tools for thaumatin:taste receptor interaction studies.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" IndexingMethod="Curated" Owner="NLM"><PMID Version="1">28764073</PMID><DateCompleted><Year>2017</Year><Month>10</Month><Day>31</Day></DateCompleted><DateRevised><Year>2018</Year><Month>12</Month><Day>02</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1873-7072</ISSN><JournalIssue CitedMedium="Internet"><Volume>237</Volume><PubDate><Year>2017</Year><Month>Dec</Month><Day>15</Day></PubDate></JournalIssue><Title>Food chemistry</Title><ISOAbbreviation>Food Chem</ISOAbbreviation></Journal><ArticleTitle>An improved process for the production of highly purified recombinant thaumatin tagged-variants.</ArticleTitle><Pagination><MedlinePgn>825-832</MedlinePgn></Pagination><ELocationID EIdType="pii" ValidYN="Y">S0308-8146(17)31001-4</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.foodchem.2017.06.018</ELocationID><Abstract><AbstractText>The sweetest tasting molecule known is the protein thaumatin, first isolated from the katemfe fruit, Thaumatococcus daniellii. Thaumatin is used in the food and beverage industry as a low-calorie sugar substitute. Thaumatin interacts with taste receptors in the oral cavity eliciting a persistent sweet taste and a bitter, liquorice flavor. Recombinant thaumatin was expressed in Pichia pastoris and through a co-expression strategy with a molecular chaperone, yields of one engineered thaumatin variant increased by greater than two-fold. A detailed purification strategy for thaumatin is reported resulting in a homogenous sample recovered at a yield of 42%. The recombinant thaumatins were extensively characterised using size exclusion chromatography for homogeneity, reversed-phase HPLC for purity (99%), peptide digest LC-MS/MS for sequence determination, and circular dichroism and tryptophan fluorescence spectroscopies for conformational characterisation. These new thaumatin variants are amenable for bioconjugation, providing chemical biology tools for thaumatin:taste receptor interaction studies.</AbstractText><CopyrightInformation>Copyright © 2017 Elsevier Ltd. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Healey</LastName><ForeName>Robert D</ForeName><Initials>RD</Initials><AffiliationInfo><Affiliation>School of Chemistry, The Australian Centre for Nanomedicine and the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney, NSW, Australia. Electronic address: robert.healey@unsw.edu.au.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Lebhar</LastName><ForeName>Helene</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia. Electronic address: h.lebhar@unsw.edu.au.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hornung</LastName><ForeName>Simon</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia. Electronic address: si.hornung@web.de.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Thordarson</LastName><ForeName>Pall</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>School of Chemistry, The Australian Centre for Nanomedicine and the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney, NSW, Australia. Electronic address: p.thordarson@unsw.edu.au.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Marquis</LastName><ForeName>Christopher P</ForeName><Initials>CP</Initials><AffiliationInfo><Affiliation>School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia. Electronic address: c.marquis@unsw.edu.au.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2017</Year><Month>06</Month><Day>13</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Food Chem</MedlineTA><NlmUniqueID>7702639</NlmUniqueID><ISSNLinking>0308-8146</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010940">Plant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D013549">Sweetening Agents</NameOfSubstance></Chemical><Chemical><RegistryNumber>53850-34-3</RegistryNumber><NameOfSubstance UI="C003427">thaumatin protein, plant</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D032426" MajorTopicYN="N">Marantaceae</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010843" MajorTopicYN="N">Pichia</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010940" MajorTopicYN="N">Plant Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013549" MajorTopicYN="N">Sweetening Agents</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D053719" MajorTopicYN="N">Tandem Mass Spectrometry</DescriptorName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Fermentation</Keyword><Keyword MajorTopicYN="N">GPCR</Keyword><Keyword MajorTopicYN="N">Pichia pastoris</Keyword><Keyword MajorTopicYN="N">Protein disulfide isomerase</Keyword><Keyword MajorTopicYN="N">Sweet tasting protein</Keyword><Keyword MajorTopicYN="N">Sweetener</Keyword><Keyword MajorTopicYN="N">Thaumatin</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2017</Year><Month>02</Month><Day>28</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2017</Year><Month>05</Month><Day>03</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2017</Year><Month>06</Month><Day>05</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2017</Year><Month>8</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2017</Year><Month>8</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2017</Year><Month>11</Month><Day>1</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">28764073</ArticleId><ArticleId IdType="pii">S0308-8146(17)31001-4</ArticleId><ArticleId IdType="doi">10.1016/j.foodchem.2017.06.018</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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