Site-directed mutagenesis of manganese peroxidase from Phanerochaete chrysosporium in an in vitro expression system.
Identifieur interne : 000603 ( Main/Exploration ); précédent : 000602; suivant : 000604Site-directed mutagenesis of manganese peroxidase from Phanerochaete chrysosporium in an in vitro expression system.
Auteurs : Xiaomei Zhang [République populaire de Chine] ; Yuxiao Wang ; Lushan Wang ; Guanjun Chen ; Weifeng Liu ; Peiji GaoSource :
- Journal of biotechnology [ 0168-1656 ] ; 2009.
Descripteurs français
- KwdFr :
- Cinétique (MeSH), Domaine catalytique (génétique), Modèles moléculaires (MeSH), Mutagenèse dirigée (MeSH), Peroxidases (composition chimique), Peroxidases (génétique), Peroxidases (métabolisme), Phanerochaete (enzymologie), Phanerochaete (génétique), Protéines fongiques (composition chimique), Protéines fongiques (génétique), Protéines fongiques (métabolisme), Pyrogallol (analogues et dérivés), Pyrogallol (métabolisme), Simulation numérique (MeSH), Sites de fixation (génétique).
- MESH :
- analogues et dérivés : Pyrogallol.
- composition chimique : Peroxidases, Protéines fongiques.
- enzymologie : Phanerochaete.
- génétique : Domaine catalytique, Peroxidases, Phanerochaete, Protéines fongiques, Sites de fixation.
- métabolisme : Peroxidases, Protéines fongiques, Pyrogallol.
- Cinétique, Modèles moléculaires, Mutagenèse dirigée, Simulation numérique.
English descriptors
- KwdEn :
- Binding Sites (genetics), Catalytic Domain (genetics), Computer Simulation (MeSH), Fungal Proteins (chemistry), Fungal Proteins (genetics), Fungal Proteins (metabolism), Kinetics (MeSH), Models, Molecular (MeSH), Mutagenesis, Site-Directed (MeSH), Peroxidases (chemistry), Peroxidases (genetics), Peroxidases (metabolism), Phanerochaete (enzymology), Phanerochaete (genetics), Pyrogallol (analogs & derivatives), Pyrogallol (metabolism).
- MESH :
- chemical , analogs & derivatives : Pyrogallol.
- chemical , chemistry : Fungal Proteins, Peroxidases.
- enzymology : Phanerochaete.
- genetics : Binding Sites, Catalytic Domain, Fungal Proteins, Peroxidases, Phanerochaete.
- chemical , metabolism : Fungal Proteins, Peroxidases, Pyrogallol.
- Computer Simulation, Kinetics, Models, Molecular, Mutagenesis, Site-Directed.
Abstract
An enzymatically active fungal manganese peroxidase (MnP) from Phanerochaete chrysosporium was synthesized in an in vitro coupled transcription-translation system. The synthesized MnP had the expected molecular mass (43,000Da) and catalyzed the oxidation of 2,6-dimethoxyphenol (DMP) in the presence of hydrogen peroxide and Mn(II). A distal arginine residue (Arg 42) of the peroxide binding pocket and the potential N-glycosylation site (Asn 131) was site-directed mutagenized and corresponding mutant enzymes were also in vitro synthesized. Activities of the mutant enzymes towards 2,6-DMP were not significantly compromised although their dynamic characteristics were obviously different from the wild-type enzyme. The effect of the mutations was explained by using a computer-based three-dimensional modeling. These results demonstrated that in vitro expression of MnP provided a convenient and efficient system for characterization of fungal peroxidases.
DOI: 10.1016/j.jbiotec.2008.10.006
PubMed: 19027802
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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first="Lushan" last="Wang">Lushan Wang</name></author><author><name sortKey="Chen, Guanjun" sort="Chen, Guanjun" uniqKey="Chen G" first="Guanjun" last="Chen">Guanjun Chen</name></author><author><name sortKey="Liu, Weifeng" sort="Liu, Weifeng" uniqKey="Liu W" first="Weifeng" last="Liu">Weifeng Liu</name></author><author><name sortKey="Gao, Peiji" sort="Gao, Peiji" uniqKey="Gao P" first="Peiji" last="Gao">Peiji Gao</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2009">2009</date><idno type="RBID">pubmed:19027802</idno><idno type="pmid">19027802</idno><idno type="doi">10.1016/j.jbiotec.2008.10.006</idno><idno type="wicri:Area/Main/Corpus">000656</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000656</idno><idno type="wicri:Area/Main/Curation">000656</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000656</idno><idno 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type="abstract" xml:lang="en">An enzymatically active fungal manganese peroxidase (MnP) from Phanerochaete chrysosporium was synthesized in an in vitro coupled transcription-translation system. The synthesized MnP had the expected molecular mass (43,000Da) and catalyzed the oxidation of 2,6-dimethoxyphenol (DMP) in the presence of hydrogen peroxide and Mn(II). A distal arginine residue (Arg 42) of the peroxide binding pocket and the potential N-glycosylation site (Asn 131) was site-directed mutagenized and corresponding mutant enzymes were also in vitro synthesized. Activities of the mutant enzymes towards 2,6-DMP were not significantly compromised although their dynamic characteristics were obviously different from the wild-type enzyme. The effect of the mutations was explained by using a computer-based three-dimensional modeling. These results demonstrated that in vitro expression of MnP provided a convenient and efficient system for characterization of fungal peroxidases.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">19027802</PMID><DateCompleted><Year>2009</Year><Month>07</Month><Day>31</Day></DateCompleted><DateRevised><Year>2015</Year><Month>11</Month><Day>19</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">0168-1656</ISSN><JournalIssue CitedMedium="Print"><Volume>139</Volume><Issue>2</Issue><PubDate><Year>2009</Year><Month>Jan</Month><Day>15</Day></PubDate></JournalIssue><Title>Journal of biotechnology</Title><ISOAbbreviation>J Biotechnol</ISOAbbreviation></Journal><ArticleTitle>Site-directed mutagenesis of manganese peroxidase from Phanerochaete chrysosporium in an in vitro expression system.</ArticleTitle><Pagination><MedlinePgn>176-8</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.jbiotec.2008.10.006</ELocationID><Abstract><AbstractText>An enzymatically active fungal manganese peroxidase (MnP) from Phanerochaete chrysosporium was synthesized in an in vitro coupled transcription-translation system. The synthesized MnP had the expected molecular mass (43,000Da) and catalyzed the oxidation of 2,6-dimethoxyphenol (DMP) in the presence of hydrogen peroxide and Mn(II). A distal arginine residue (Arg 42) of the peroxide binding pocket and the potential N-glycosylation site (Asn 131) was site-directed mutagenized and corresponding mutant enzymes were also in vitro synthesized. Activities of the mutant enzymes towards 2,6-DMP were not significantly compromised although their dynamic characteristics were obviously different from the wild-type enzyme. The effect of the mutations was explained by using a computer-based three-dimensional modeling. These results demonstrated that in vitro expression of MnP provided a convenient and efficient system for characterization of fungal peroxidases.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Xiaomei</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>The State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Jinan 250100, Shandong, PR China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Yuxiao</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Lushan</ForeName><Initials>L</Initials></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>Guanjun</ForeName><Initials>G</Initials></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Weifeng</ForeName><Initials>W</Initials></Author><Author ValidYN="Y"><LastName>Gao</LastName><ForeName>Peiji</ForeName><Initials>P</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2008</Year><Month>11</Month><Day>05</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>J Biotechnol</MedlineTA><NlmUniqueID>8411927</NlmUniqueID><ISSNLinking>0168-1656</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D005656">Fungal Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>01Y4A2QXY0</RegistryNumber><NameOfSubstance UI="D011748">Pyrogallol</NameOfSubstance></Chemical><Chemical><RegistryNumber>4UQT464H8K</RegistryNumber><NameOfSubstance UI="C010120">pyrogallol 1,3-dimethyl ether</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="D001665" MajorTopicYN="N">Binding Sites</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020134" MajorTopicYN="N">Catalytic Domain</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D003198" MajorTopicYN="N">Computer Simulation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005656" MajorTopicYN="N">Fungal Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D007700" MajorTopicYN="N">Kinetics</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008958" MajorTopicYN="N">Models, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D016297" MajorTopicYN="Y">Mutagenesis, Site-Directed</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010544" MajorTopicYN="N">Peroxidases</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020075" MajorTopicYN="N">Phanerochaete</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011748" MajorTopicYN="N">Pyrogallol</DescriptorName><QualifierName UI="Q000031" MajorTopicYN="N">analogs & derivatives</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2008</Year><Month>06</Month><Day>12</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2008</Year><Month>10</Month><Day>15</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2008</Year><Month>10</Month><Day>21</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2008</Year><Month>11</Month><Day>26</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2008</Year><Month>11</Month><Day>26</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2009</Year><Month>8</Month><Day>1</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">19027802</ArticleId><ArticleId IdType="pii">S0168-1656(08)02411-5</ArticleId><ArticleId IdType="doi">10.1016/j.jbiotec.2008.10.006</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><noCountry><name sortKey="Chen, Guanjun" sort="Chen, Guanjun" uniqKey="Chen G" first="Guanjun" last="Chen">Guanjun Chen</name><name sortKey="Gao, Peiji" sort="Gao, Peiji" uniqKey="Gao P" first="Peiji" last="Gao">Peiji Gao</name><name sortKey="Liu, Weifeng" sort="Liu, Weifeng" uniqKey="Liu W" first="Weifeng" last="Liu">Weifeng Liu</name><name sortKey="Wang, Lushan" sort="Wang, Lushan" uniqKey="Wang L" first="Lushan" last="Wang">Lushan Wang</name><name sortKey="Wang, Yuxiao" sort="Wang, Yuxiao" uniqKey="Wang Y" first="Yuxiao" last="Wang">Yuxiao Wang</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Zhang, Xiaomei" sort="Zhang, Xiaomei" uniqKey="Zhang X" first="Xiaomei" last="Zhang">Xiaomei Zhang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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