Functional Expression and One-Step Protein Purification of Manganese Peroxidase 1 (rMnP1) from Phanerochaetechrysosporium Using the E. coli-Expression System.
Identifieur interne : 000088 ( Main/Exploration ); précédent : 000087; suivant : 000089Functional Expression and One-Step Protein Purification of Manganese Peroxidase 1 (rMnP1) from Phanerochaetechrysosporium Using the E. coli-Expression System.
Auteurs : Angel De La Cruz Pech-Canul [Mexique] ; Javier Carrillo-Campos [Mexique] ; María De Lourdes Ballinas-Casarrubias [Mexique] ; Rosa Lidia Solis-Oviedo [Mexique] ; Selena Karina Hernández-Rasc N [Mexique] ; Le N Raúl Hernández-Ochoa [Mexique] ; Néstor Gutiérrez-Méndez [Mexique] ; Antonio García-Triana [Mexique]Source :
- International journal of molecular sciences [ 1422-0067 ] ; 2020.
Descripteurs français
- KwdFr :
- Dosages enzymatiques (MeSH), Escherichia coli (métabolisme), Expression des gènes (MeSH), Normes de référence (MeSH), Peroxidases (isolement et purification), Phanerochaete (enzymologie), Protéines de fusion recombinantes (génétique), Protéines de fusion recombinantes (isolement et purification), Protéines de fusion recombinantes (métabolisme), Solubilité (MeSH), Séquence d'acides aminés (MeSH), Vecteurs génétiques (métabolisme).
- MESH :
- enzymologie : Phanerochaete.
- génétique : Protéines de fusion recombinantes.
- isolement et purification : Peroxidases, Protéines de fusion recombinantes.
- métabolisme : Escherichia coli, Protéines de fusion recombinantes, Vecteurs génétiques.
- Dosages enzymatiques, Expression des gènes, Normes de référence, Solubilité, Séquence d'acides aminés.
English descriptors
- KwdEn :
- Amino Acid Sequence (MeSH), Enzyme Assays (MeSH), Escherichia coli (metabolism), Gene Expression (MeSH), Genetic Vectors (metabolism), Peroxidases (isolation & purification), Phanerochaete (enzymology), Recombinant Fusion Proteins (genetics), Recombinant Fusion Proteins (isolation & purification), Recombinant Fusion Proteins (metabolism), Reference Standards (MeSH), Solubility (MeSH).
- MESH :
- chemical , genetics : Recombinant Fusion Proteins.
- chemical , isolation & purification : Peroxidases, Recombinant Fusion Proteins.
- enzymology : Phanerochaete.
- metabolism : Escherichia coli, Genetic Vectors, Recombinant Fusion Proteins.
- Amino Acid Sequence, Enzyme Assays, Gene Expression, Reference Standards, Solubility.
Abstract
Manganese peroxidases (MnP) from the white-rot fungi Phanerochaete chrysosporium catalyse the oxidation of Mn2+ to Mn3+, a strong oxidizer able to oxidize a wide variety of organic compounds. Different approaches have been used to unravel the enzymatic properties and potential applications of MnP. However, these efforts have been hampered by the limited production of native MnP by fungi. Heterologous expression of MnP has been achieved in both eukaryotic and prokaryotic expression systems, although with limited production and many disadvantages in the process. Here we described a novel molecular approach for the expression and purification of manganese peroxidase isoform 1 (MnP1) from P. chrysosporium using an E. coli-expression system. The proposed strategy involved the codon optimization and chemical synthesis of the MnP1 gene for optimised expression in the E. coli T7 shuffle host. Recombinant MnP1 (rMnP1) was expressed as a fusion protein, which was recovered from solubilised inclusion bodies. rMnP1 was purified from the fusion protein using intein-based protein purification techniques and a one-step affinity chromatography. The designated strategy allowed production of an active enzyme able to oxidize guaiacol or Mn2+.
DOI: 10.3390/ijms21020416
PubMed: 31936493
PubMed Central: PMC7013543
Affiliations:
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Le document en format XML
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<i>Phanerochaete</i><i>chrysosporium</i> Using the <i>E. coli</i>-Expression System.</title><author><name sortKey="Pech Canul, Angel De La Cruz" sort="Pech Canul, Angel De La Cruz" uniqKey="Pech Canul A" first="Angel De La Cruz" last="Pech-Canul">Angel De La Cruz Pech-Canul</name><affiliation wicri:level="1"><nlm:affiliation>CONACyT-Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</nlm:affiliation><country xml:lang="fr">Mexique</country><wicri:regionArea>CONACyT-Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125</wicri:regionArea><wicri:noRegion>Chihuahua 31125</wicri:noRegion></affiliation></author><author><name sortKey="Carrillo Campos, Javier" sort="Carrillo Campos, Javier" uniqKey="Carrillo Campos J" first="Javier" last="Carrillo-Campos">Javier Carrillo-Campos</name><affiliation wicri:level="1"><nlm:affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</nlm:affiliation><country xml:lang="fr">Mexique</country><wicri:regionArea>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125</wicri:regionArea><wicri:noRegion>Chihuahua 31125</wicri:noRegion></affiliation></author><author><name sortKey="Ballinas Casarrubias, Maria De Lourdes" sort="Ballinas Casarrubias, Maria De Lourdes" uniqKey="Ballinas Casarrubias M" first="María De Lourdes" last="Ballinas-Casarrubias">María De Lourdes Ballinas-Casarrubias</name><affiliation wicri:level="1"><nlm:affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</nlm:affiliation><country xml:lang="fr">Mexique</country><wicri:regionArea>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125</wicri:regionArea><wicri:noRegion>Chihuahua 31125</wicri:noRegion></affiliation></author><author><name sortKey="Solis Oviedo, Rosa Lidia" sort="Solis Oviedo, Rosa Lidia" uniqKey="Solis Oviedo R" first="Rosa Lidia" last="Solis-Oviedo">Rosa Lidia Solis-Oviedo</name><affiliation wicri:level="1"><nlm:affiliation>Healthy Life Importadora, Exportadora Y Productora De Alimentos Y Bebidas, Sociedad De Responsabilidad Limitada De Capital Variable, Chihuahua 31125, Mexico.</nlm:affiliation><country xml:lang="fr">Mexique</country><wicri:regionArea>Healthy Life Importadora, Exportadora Y Productora De Alimentos Y Bebidas, Sociedad De Responsabilidad Limitada De Capital Variable, Chihuahua 31125</wicri:regionArea><wicri:noRegion>Chihuahua 31125</wicri:noRegion></affiliation></author><author><name sortKey="Hernandez Rasc N, Selena Karina" sort="Hernandez Rasc N, Selena Karina" uniqKey="Hernandez Rasc N S" first="Selena Karina" last="Hernández-Rasc N">Selena Karina Hernández-Rasc N</name><affiliation wicri:level="1"><nlm:affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</nlm:affiliation><country 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Gutiérrez-Méndez</name><affiliation wicri:level="1"><nlm:affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</nlm:affiliation><country xml:lang="fr">Mexique</country><wicri:regionArea>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125</wicri:regionArea><wicri:noRegion>Chihuahua 31125</wicri:noRegion></affiliation></author><author><name sortKey="Garcia Triana, Antonio" sort="Garcia Triana, Antonio" uniqKey="Garcia Triana A" first="Antonio" last="García-Triana">Antonio García-Triana</name><affiliation wicri:level="1"><nlm:affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</nlm:affiliation><country xml:lang="fr">Mexique</country><wicri:regionArea>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125</wicri:regionArea><wicri:noRegion>Chihuahua 31125</wicri:noRegion></affiliation></author></analytic><series><title level="j">International journal of molecular sciences</title><idno type="eISSN">1422-0067</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence (MeSH)</term><term>Enzyme Assays (MeSH)</term><term>Escherichia coli (metabolism)</term><term>Gene Expression (MeSH)</term><term>Genetic Vectors (metabolism)</term><term>Peroxidases (isolation & purification)</term><term>Phanerochaete (enzymology)</term><term>Recombinant Fusion Proteins (genetics)</term><term>Recombinant Fusion Proteins (isolation & purification)</term><term>Recombinant Fusion Proteins (metabolism)</term><term>Reference Standards (MeSH)</term><term>Solubility (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Dosages enzymatiques (MeSH)</term><term>Escherichia coli (métabolisme)</term><term>Expression des gènes (MeSH)</term><term>Normes de référence (MeSH)</term><term>Peroxidases (isolement et purification)</term><term>Phanerochaete (enzymologie)</term><term>Protéines de fusion recombinantes (génétique)</term><term>Protéines de fusion recombinantes (isolement et purification)</term><term>Protéines de fusion recombinantes (métabolisme)</term><term>Solubilité (MeSH)</term><term>Séquence d'acides aminés (MeSH)</term><term>Vecteurs génétiques (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Recombinant Fusion Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Peroxidases</term><term>Recombinant Fusion Proteins</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines de fusion recombinantes</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Peroxidases</term><term>Protéines de fusion recombinantes</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Escherichia coli</term><term>Genetic Vectors</term><term>Recombinant Fusion Proteins</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Escherichia coli</term><term>Protéines de fusion recombinantes</term><term>Vecteurs génétiques</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Enzyme Assays</term><term>Gene Expression</term><term>Reference Standards</term><term>Solubility</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Dosages enzymatiques</term><term>Expression des gènes</term><term>Normes de référence</term><term>Solubilité</term><term>Séquence d'acides aminés</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Manganese peroxidases (MnP) from the white-rot fungi <i>Phanerochaete chrysosporium</i> catalyse the oxidation of Mn<sup>2+</sup> to Mn<sup>3+</sup>, a strong oxidizer able to oxidize a wide variety of organic compounds. Different approaches have been used to unravel the enzymatic properties and potential applications of MnP. However, these efforts have been hampered by the limited production of native MnP by fungi. Heterologous expression of MnP has been achieved in both eukaryotic and prokaryotic expression systems, although with limited production and many disadvantages in the process. Here we described a novel molecular approach for the expression and purification of manganese peroxidase isoform 1 (MnP1) from <i>P. chrysosporium</i> using an <i>E. coli</i>-expression system. The proposed strategy involved the codon optimization and chemical synthesis of the MnP1 gene for optimised expression in the <i>E. coli</i> T7 shuffle host. Recombinant MnP1 (rMnP1) was expressed as a fusion protein, which was recovered from solubilised inclusion bodies. rMnP1 was purified from the fusion protein using intein-based protein purification techniques and a one-step affinity chromatography. The designated strategy allowed production of an active enzyme able to oxidize guaiacol or Mn<sup>2+</sup>.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">31936493</PMID><DateCompleted><Year>2020</Year><Month>09</Month><Day>30</Day></DateCompleted><DateRevised><Year>2020</Year><Month>09</Month><Day>30</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1422-0067</ISSN><JournalIssue CitedMedium="Internet"><Volume>21</Volume><Issue>2</Issue><PubDate><Year>2020</Year><Month>Jan</Month><Day>09</Day></PubDate></JournalIssue><Title>International journal of molecular sciences</Title><ISOAbbreviation>Int J Mol Sci</ISOAbbreviation></Journal><ArticleTitle>Functional Expression and One-Step Protein Purification of Manganese Peroxidase 1 (rMnP1) from <i>Phanerochaete</i><i>chrysosporium</i> Using the <i>E. coli</i>-Expression System.</ArticleTitle><ELocationID EIdType="pii" ValidYN="Y">E416</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.3390/ijms21020416</ELocationID><Abstract><AbstractText>Manganese peroxidases (MnP) from the white-rot fungi <i>Phanerochaete chrysosporium</i> catalyse the oxidation of Mn<sup>2+</sup> to Mn<sup>3+</sup>, a strong oxidizer able to oxidize a wide variety of organic compounds. Different approaches have been used to unravel the enzymatic properties and potential applications of MnP. However, these efforts have been hampered by the limited production of native MnP by fungi. Heterologous expression of MnP has been achieved in both eukaryotic and prokaryotic expression systems, although with limited production and many disadvantages in the process. Here we described a novel molecular approach for the expression and purification of manganese peroxidase isoform 1 (MnP1) from <i>P. chrysosporium</i> using an <i>E. coli</i>-expression system. The proposed strategy involved the codon optimization and chemical synthesis of the MnP1 gene for optimised expression in the <i>E. coli</i> T7 shuffle host. Recombinant MnP1 (rMnP1) was expressed as a fusion protein, which was recovered from solubilised inclusion bodies. rMnP1 was purified from the fusion protein using intein-based protein purification techniques and a one-step affinity chromatography. The designated strategy allowed production of an active enzyme able to oxidize guaiacol or Mn<sup>2+</sup>.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Pech-Canul</LastName><ForeName>Angel De La Cruz</ForeName><Initials>AC</Initials><Identifier Source="ORCID">0000-0002-1874-322X</Identifier><AffiliationInfo><Affiliation>CONACyT-Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Carrillo-Campos</LastName><ForeName>Javier</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ballinas-Casarrubias</LastName><ForeName>María de Lourdes</ForeName><Initials>ML</Initials><AffiliationInfo><Affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Solis-Oviedo</LastName><ForeName>Rosa Lidia</ForeName><Initials>RL</Initials><Identifier Source="ORCID">0000-0001-9801-5283</Identifier><AffiliationInfo><Affiliation>Healthy Life Importadora, Exportadora Y Productora De Alimentos Y Bebidas, Sociedad De Responsabilidad Limitada De Capital Variable, Chihuahua 31125, Mexico.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hernández-Rascón</LastName><ForeName>Selena Karina</ForeName><Initials>SK</Initials><AffiliationInfo><Affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hernández-Ochoa</LastName><ForeName>León Raúl</ForeName><Initials>LR</Initials><AffiliationInfo><Affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Gutiérrez-Méndez</LastName><ForeName>Néstor</ForeName><Initials>N</Initials><Identifier Source="ORCID">0000-0002-0791-3211</Identifier><AffiliationInfo><Affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>García-Triana</LastName><ForeName>Antonio</ForeName><Initials>A</Initials><Identifier Source="ORCID">0000-0002-4762-9135</Identifier><AffiliationInfo><Affiliation>Faculty of Chemical Sciences, Autonomous University of Chihuahua, Campus II, Chihuahua 31125, Mexico.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>CB-2011-01-000000000167036</GrantID><Agency>Consejo Nacional de Ciencia y Tecnología</Agency><Country></Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>01</Month><Day>09</Day></ArticleDate></Article><MedlineJournalInfo><Country>Switzerland</Country><MedlineTA>Int J Mol Sci</MedlineTA><NlmUniqueID>101092791</NlmUniqueID><ISSNLinking>1422-0067</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011993">Recombinant Fusion Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.11.1.-</RegistryNumber><NameOfSubstance UI="D010544">Peroxidases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.11.1.13</RegistryNumber><NameOfSubstance UI="C051129">manganese peroxidase</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000595" MajorTopicYN="N">Amino Acid Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D057075" MajorTopicYN="N">Enzyme Assays</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004926" MajorTopicYN="N">Escherichia coli</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015870" MajorTopicYN="Y">Gene Expression</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005822" MajorTopicYN="N">Genetic Vectors</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010544" MajorTopicYN="N">Peroxidases</DescriptorName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020075" MajorTopicYN="N">Phanerochaete</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011993" MajorTopicYN="N">Recombinant Fusion Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012015" MajorTopicYN="N">Reference Standards</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012995" MajorTopicYN="N">Solubility</DescriptorName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Escherichia coli</Keyword><Keyword MajorTopicYN="N">MnP1</Keyword><Keyword MajorTopicYN="N">Phanerochaete</Keyword><Keyword MajorTopicYN="N">affinity chromatography</Keyword><Keyword MajorTopicYN="N">chitin binding domain (CBD)</Keyword><Keyword MajorTopicYN="N">codon-optimization</Keyword><Keyword MajorTopicYN="N">inteins</Keyword><Keyword MajorTopicYN="N">manganese peroxidase</Keyword><Keyword MajorTopicYN="N">protein purification</Keyword><Keyword MajorTopicYN="N">recombinant</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>12</Month><Day>03</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2019</Year><Month>12</Month><Day>14</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2019</Year><Month>12</Month><Day>19</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>1</Month><Day>16</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>1</Month><Day>16</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>10</Month><Day>2</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">31936493</ArticleId><ArticleId IdType="pii">ijms21020416</ArticleId><ArticleId 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IdType="pubmed">27908283</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed><affiliations><list><country><li>Mexique</li></country></list><tree><country name="Mexique"><noRegion><name sortKey="Pech Canul, Angel De La Cruz" sort="Pech Canul, Angel De La Cruz" uniqKey="Pech Canul A" first="Angel De La Cruz" last="Pech-Canul">Angel De La Cruz Pech-Canul</name></noRegion><name sortKey="Ballinas Casarrubias, Maria De Lourdes" sort="Ballinas Casarrubias, Maria De Lourdes" uniqKey="Ballinas Casarrubias M" first="María De Lourdes" last="Ballinas-Casarrubias">María De Lourdes Ballinas-Casarrubias</name><name sortKey="Carrillo Campos, Javier" sort="Carrillo Campos, Javier" uniqKey="Carrillo Campos J" first="Javier" last="Carrillo-Campos">Javier Carrillo-Campos</name><name sortKey="Garcia Triana, Antonio" sort="Garcia Triana, Antonio" uniqKey="Garcia Triana A" first="Antonio" last="García-Triana">Antonio García-Triana</name><name sortKey="Gutierrez Mendez, Nestor" sort="Gutierrez Mendez, Nestor" uniqKey="Gutierrez Mendez N" first="Néstor" last="Gutiérrez-Méndez">Néstor Gutiérrez-Méndez</name><name sortKey="Hernandez Ochoa, Le N Raul" sort="Hernandez Ochoa, Le N Raul" uniqKey="Hernandez Ochoa L" first="Le N Raúl" last="Hernández-Ochoa">Le N Raúl Hernández-Ochoa</name><name sortKey="Hernandez Rasc N, Selena Karina" sort="Hernandez Rasc N, Selena Karina" uniqKey="Hernandez Rasc N S" first="Selena Karina" last="Hernández-Rasc N">Selena Karina Hernández-Rasc N</name><name sortKey="Solis Oviedo, Rosa Lidia" sort="Solis Oviedo, Rosa Lidia" uniqKey="Solis Oviedo R" first="Rosa Lidia" last="Solis-Oviedo">Rosa Lidia Solis-Oviedo</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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