Pyranose 2-oxidase from Phanerochaete chrysosporium--further biochemical characterisation.
Identifieur interne : 000C00 ( Main/Exploration ); précédent : 000B99; suivant : 000C01Pyranose 2-oxidase from Phanerochaete chrysosporium--further biochemical characterisation.
Auteurs : M J Artolozaga [Allemagne] ; E. Kubátová ; J. Volc ; H M KaliszSource :
- Applied microbiology and biotechnology [ 0175-7598 ] ; 1997.
Descripteurs français
- KwdFr :
- MESH :
- enzymologie : Basidiomycota.
- isolement et purification : Carbohydrate dehydrogenases.
- métabolisme : Carbohydrate dehydrogenases.
- Cinétique, Concentration en ions d'hydrogène, Température.
English descriptors
- KwdEn :
- MESH :
- chemical , isolation & purification : Carbohydrate Dehydrogenases.
- enzymology : Basidiomycota.
- chemical , metabolism : Carbohydrate Dehydrogenases.
- Hydrogen-Ion Concentration, Kinetics, Temperature.
Abstract
Pyranose 2-oxidase (P2O) was purified 43-fold to apparent homogeneity from the basidiomycete Phanerochaete chrysosporium using liquid chromatography on phenyl Sepharose, Mono Q (twice) and phenyl Superose. The native enzyme has a molecular mass of about 250 kDa (based on native PAGE) and is composed of four identical subunits of 65 kDa. It contains three isoforms of isoelectric point (pI) 5.0, 5.05 and 5.15 and does not appear to be a glycoprotein. P2O is optimally stable at pH 8.0 and up to 60 degrees C. It is active over a broad pH range (5.0-9.0) with maximum activity at pH 8.0-8.5 and at 55 degrees C, and a broad substrate specificity. D-Glucose is the preferred substrate, but 1-beta-aurothioglucose, 6-deoxy-D-glucose, L-sorbose, D-xylose, 5-thioglucose, D-glucono-1,5-lactone, maltose and 2-deoxy-D-glucose are also oxidised at relatively high rates. A Ping Pong Bi Bi mechanism was demonstrated for the P2O reaction at pH 8.0, with a catalytic constant (kcat) of 111.0 s-1 and an affinity constant (Km) of 1.43 mM for D-glucose and 83.2 microM for oxygen. Whereas the steady-state kinetics for glucose oxidation were unaffected by the medium at pH > or = 7.0, at low pH both pH and buffer composition affected the P2O kinetics with the kcat/K(m) value decreasing with decreasing pH. The greatest effect was observed in acetate buffer (0.1 M, pH 4.5), where the kcat decreased to 60.9 s-1 and the K(m) increased to 240 mM. The activity of P2O was completely inhibited by 10 mM HgCl2, AgNO3 and ZnCl2, and 50% by lead acetate, CuCl2 and MnCl2.
DOI: 10.1007/s002530050964
PubMed: 9210340
Affiliations:
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Volc</name></author><author><name sortKey="Kalisz, H M" sort="Kalisz, H M" uniqKey="Kalisz H" first="H M" last="Kalisz">H M Kalisz</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1997">1997</date><idno type="RBID">pubmed:9210340</idno><idno type="pmid">9210340</idno><idno type="doi">10.1007/s002530050964</idno><idno type="wicri:Area/Main/Corpus">000C13</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000C13</idno><idno type="wicri:Area/Main/Curation">000C13</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000C13</idno><idno type="wicri:Area/Main/Exploration">000C13</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Pyranose 2-oxidase from Phanerochaete chrysosporium--further biochemical characterisation.</title><author><name sortKey="Artolozaga, M J" sort="Artolozaga, M J" uniqKey="Artolozaga M" first="M J" last="Artolozaga">M J Artolozaga</name><affiliation wicri:level="1"><nlm:affiliation>GBF-Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>GBF-Gesellschaft für Biotechnologische Forschung, Braunschweig</wicri:regionArea><wicri:noRegion>Braunschweig</wicri:noRegion><wicri:noRegion>Braunschweig</wicri:noRegion><wicri:noRegion>Braunschweig</wicri:noRegion></affiliation></author><author><name sortKey="Kubatova, E" sort="Kubatova, E" uniqKey="Kubatova E" first="E" last="Kubátová">E. Kubátová</name></author><author><name sortKey="Volc, J" sort="Volc, J" uniqKey="Volc J" first="J" last="Volc">J. Volc</name></author><author><name sortKey="Kalisz, H M" sort="Kalisz, H M" uniqKey="Kalisz H" first="H M" last="Kalisz">H M Kalisz</name></author></analytic><series><title level="j">Applied microbiology and biotechnology</title><idno type="ISSN">0175-7598</idno><imprint><date when="1997" type="published">1997</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Basidiomycota (enzymology)</term><term>Carbohydrate Dehydrogenases (isolation & purification)</term><term>Carbohydrate Dehydrogenases (metabolism)</term><term>Hydrogen-Ion Concentration (MeSH)</term><term>Kinetics (MeSH)</term><term>Temperature (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Basidiomycota (enzymologie)</term><term>Carbohydrate dehydrogenases (isolement et purification)</term><term>Carbohydrate dehydrogenases (métabolisme)</term><term>Cinétique (MeSH)</term><term>Concentration en ions d'hydrogène (MeSH)</term><term>Température (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Carbohydrate Dehydrogenases</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Basidiomycota</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Basidiomycota</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Carbohydrate dehydrogenases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Carbohydrate Dehydrogenases</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Carbohydrate dehydrogenases</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Hydrogen-Ion Concentration</term><term>Kinetics</term><term>Temperature</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Cinétique</term><term>Concentration en ions d'hydrogène</term><term>Température</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Pyranose 2-oxidase (P2O) was purified 43-fold to apparent homogeneity from the basidiomycete Phanerochaete chrysosporium using liquid chromatography on phenyl Sepharose, Mono Q (twice) and phenyl Superose. The native enzyme has a molecular mass of about 250 kDa (based on native PAGE) and is composed of four identical subunits of 65 kDa. It contains three isoforms of isoelectric point (pI) 5.0, 5.05 and 5.15 and does not appear to be a glycoprotein. P2O is optimally stable at pH 8.0 and up to 60 degrees C. It is active over a broad pH range (5.0-9.0) with maximum activity at pH 8.0-8.5 and at 55 degrees C, and a broad substrate specificity. D-Glucose is the preferred substrate, but 1-beta-aurothioglucose, 6-deoxy-D-glucose, L-sorbose, D-xylose, 5-thioglucose, D-glucono-1,5-lactone, maltose and 2-deoxy-D-glucose are also oxidised at relatively high rates. A Ping Pong Bi Bi mechanism was demonstrated for the P2O reaction at pH 8.0, with a catalytic constant (kcat) of 111.0 s-1 and an affinity constant (Km) of 1.43 mM for D-glucose and 83.2 microM for oxygen. Whereas the steady-state kinetics for glucose oxidation were unaffected by the medium at pH > or = 7.0, at low pH both pH and buffer composition affected the P2O kinetics with the kcat/K(m) value decreasing with decreasing pH. The greatest effect was observed in acetate buffer (0.1 M, pH 4.5), where the kcat decreased to 60.9 s-1 and the K(m) increased to 240 mM. The activity of P2O was completely inhibited by 10 mM HgCl2, AgNO3 and ZnCl2, and 50% by lead acetate, CuCl2 and MnCl2.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">9210340</PMID><DateCompleted><Year>1997</Year><Month>07</Month><Day>24</Day></DateCompleted><DateRevised><Year>2019</Year><Month>09</Month><Day>14</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0175-7598</ISSN><JournalIssue CitedMedium="Print"><Volume>47</Volume><Issue>5</Issue><PubDate><Year>1997</Year><Month>May</Month></PubDate></JournalIssue><Title>Applied microbiology and biotechnology</Title><ISOAbbreviation>Appl Microbiol Biotechnol</ISOAbbreviation></Journal><ArticleTitle>Pyranose 2-oxidase from Phanerochaete chrysosporium--further biochemical characterisation.</ArticleTitle><Pagination><MedlinePgn>508-14</MedlinePgn></Pagination><Abstract><AbstractText>Pyranose 2-oxidase (P2O) was purified 43-fold to apparent homogeneity from the basidiomycete Phanerochaete chrysosporium using liquid chromatography on phenyl Sepharose, Mono Q (twice) and phenyl Superose. The native enzyme has a molecular mass of about 250 kDa (based on native PAGE) and is composed of four identical subunits of 65 kDa. It contains three isoforms of isoelectric point (pI) 5.0, 5.05 and 5.15 and does not appear to be a glycoprotein. P2O is optimally stable at pH 8.0 and up to 60 degrees C. It is active over a broad pH range (5.0-9.0) with maximum activity at pH 8.0-8.5 and at 55 degrees C, and a broad substrate specificity. D-Glucose is the preferred substrate, but 1-beta-aurothioglucose, 6-deoxy-D-glucose, L-sorbose, D-xylose, 5-thioglucose, D-glucono-1,5-lactone, maltose and 2-deoxy-D-glucose are also oxidised at relatively high rates. A Ping Pong Bi Bi mechanism was demonstrated for the P2O reaction at pH 8.0, with a catalytic constant (kcat) of 111.0 s-1 and an affinity constant (Km) of 1.43 mM for D-glucose and 83.2 microM for oxygen. Whereas the steady-state kinetics for glucose oxidation were unaffected by the medium at pH > or = 7.0, at low pH both pH and buffer composition affected the P2O kinetics with the kcat/K(m) value decreasing with decreasing pH. The greatest effect was observed in acetate buffer (0.1 M, pH 4.5), where the kcat decreased to 60.9 s-1 and the K(m) increased to 240 mM. The activity of P2O was completely inhibited by 10 mM HgCl2, AgNO3 and ZnCl2, and 50% by lead acetate, CuCl2 and MnCl2.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Artolozaga</LastName><ForeName>M J</ForeName><Initials>MJ</Initials><AffiliationInfo><Affiliation>GBF-Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kubátová</LastName><ForeName>E</ForeName><Initials>E</Initials></Author><Author ValidYN="Y"><LastName>Volc</LastName><ForeName>J</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Kalisz</LastName><ForeName>H M</ForeName><Initials>HM</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Appl Microbiol Biotechnol</MedlineTA><NlmUniqueID>8406612</NlmUniqueID><ISSNLinking>0175-7598</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>EC 1.1.-</RegistryNumber><NameOfSubstance UI="D002237">Carbohydrate Dehydrogenases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.1.3.10</RegistryNumber><NameOfSubstance UI="C022489">pyranose oxidase</NameOfSubstance></Chemical></ChemicalList><CitationSubset>B</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D001487" MajorTopicYN="N">Basidiomycota</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D002237" MajorTopicYN="N">Carbohydrate Dehydrogenases</DescriptorName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006863" MajorTopicYN="N">Hydrogen-Ion Concentration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007700" MajorTopicYN="N">Kinetics</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013696" MajorTopicYN="N">Temperature</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1997</Year><Month>5</Month><Day>1</Day></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>1997</Year><Month>5</Month><Day>1</Day><Hour>0</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>1997</Year><Month>5</Month><Day>1</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">9210340</ArticleId><ArticleId IdType="doi">10.1007/s002530050964</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Allemagne</li></country></list><tree><noCountry><name sortKey="Kalisz, H M" sort="Kalisz, H M" uniqKey="Kalisz H" first="H M" last="Kalisz">H M Kalisz</name><name sortKey="Kubatova, E" sort="Kubatova, E" uniqKey="Kubatova E" first="E" last="Kubátová">E. Kubátová</name><name sortKey="Volc, J" sort="Volc, J" uniqKey="Volc J" first="J" last="Volc">J. Volc</name></noCountry><country name="Allemagne"><noRegion><name sortKey="Artolozaga, M J" sort="Artolozaga, M J" uniqKey="Artolozaga M" first="M J" last="Artolozaga">M J Artolozaga</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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