Glyoxal oxidase of Phanerochaete chrysosporium: its characterization and activation by lignin peroxidase.
Identifieur interne : 000F66 ( Main/Corpus ); précédent : 000F65; suivant : 000F67Glyoxal oxidase of Phanerochaete chrysosporium: its characterization and activation by lignin peroxidase.
Auteurs : P J KerstenSource :
- Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 1990.
Abstract
Glyoxal oxidase (GLOX) is an extracellular H2O2-generating enzyme produced by ligninolytic cultures of Phanerochaete chrysosporium. The production, purification, and partial characterization of GLOX from agitated cultures are described here. High-oxygen levels are critical for GLOX production as for lignin peroxidase. GLOX purified by anion-exchange chromatography appears homogeneous by NaDod-SO4/PAGE (molecular mass = 68 kDa). However, analysis by isoelectric focusing indicates two major bands (pI 4.7 and 4.9) that stain as glycoproteins as well as for H2O2-producing activity in the presence of methylglyoxal. Purified GLOX shows a marked stimulation in activity when incubated with Cu2+; full activation takes more than 1 hr with 1 mM CuSO4 at pH 6. The steady-state kinetic parameters for the GLOX oxidation of methylglyoxal, glyceraldehyde, dihydroxyacetone, glycolaldehyde, acetaldehyde, glyoxal, glyoxylic acid, and formaldehyde, were determined by using a lignin peroxidase coupled-assay at pH 4.5. Of these substrates, the best is the extracellular metabolite methylglyoxal with a Km of 0.64 mM an apparent rate of catalysis, kcat, of 198 s1 under air-saturated conditions. The Km for oxygen is greater than the concentration of oxygen possible at ambient pressure--i.e., >1.3 mM at 25 degrees C. Importantly, oxygen-uptake experiments show that purified GLOX is inactive unless coupled to the peroxidase reaction. With this coupled reaction, for each mol of methylglyoxal, veratryl alcohol (a lignin peroxidase substrate), and oxygen consumed, 1 mol each of pyruvate and veratraldehyde is produced. The importance of these results is discussed in relation to the physiology of lignin biodegradation and possible extracellular regulatory mechanisms for the control of oxidase and peroxidase activities.
DOI: 10.1073/pnas.87.8.2936
PubMed: 11607073
PubMed Central: PMC53808
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Glyoxal oxidase of Phanerochaete chrysosporium: its characterization and activation by lignin peroxidase.</title><author><name sortKey="Kersten, P J" sort="Kersten, P J" uniqKey="Kersten P" first="P J" last="Kersten">P J Kersten</name><affiliation><nlm:affiliation>Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA.</nlm:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1990">1990</date><idno type="RBID">pubmed:11607073</idno><idno type="pmid">11607073</idno><idno type="pmc">PMC53808</idno><idno type="doi">10.1073/pnas.87.8.2936</idno><idno type="wicri:Area/Main/Corpus">000F66</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000F66</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Glyoxal oxidase of Phanerochaete chrysosporium: its characterization and activation by lignin peroxidase.</title><author><name sortKey="Kersten, P J" sort="Kersten, P J" uniqKey="Kersten P" first="P J" last="Kersten">P J Kersten</name><affiliation><nlm:affiliation>Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA.</nlm:affiliation></affiliation></author></analytic><series><title level="j">Proceedings of the National Academy of Sciences of the United States of America</title><idno type="ISSN">0027-8424</idno><imprint><date when="1990" type="published">1990</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Glyoxal oxidase (GLOX) is an extracellular H2O2-generating enzyme produced by ligninolytic cultures of Phanerochaete chrysosporium. The production, purification, and partial characterization of GLOX from agitated cultures are described here. High-oxygen levels are critical for GLOX production as for lignin peroxidase. GLOX purified by anion-exchange chromatography appears homogeneous by NaDod-SO4/PAGE (molecular mass = 68 kDa). However, analysis by isoelectric focusing indicates two major bands (pI 4.7 and 4.9) that stain as glycoproteins as well as for H2O2-producing activity in the presence of methylglyoxal. Purified GLOX shows a marked stimulation in activity when incubated with Cu2+; full activation takes more than 1 hr with 1 mM CuSO4 at pH 6. The steady-state kinetic parameters for the GLOX oxidation of methylglyoxal, glyceraldehyde, dihydroxyacetone, glycolaldehyde, acetaldehyde, glyoxal, glyoxylic acid, and formaldehyde, were determined by using a lignin peroxidase coupled-assay at pH 4.5. Of these substrates, the best is the extracellular metabolite methylglyoxal with a Km of 0.64 mM an apparent rate of catalysis, kcat, of 198 s1 under air-saturated conditions. The Km for oxygen is greater than the concentration of oxygen possible at ambient pressure--i.e., >1.3 mM at 25 degrees C. Importantly, oxygen-uptake experiments show that purified GLOX is inactive unless coupled to the peroxidase reaction. With this coupled reaction, for each mol of methylglyoxal, veratryl alcohol (a lignin peroxidase substrate), and oxygen consumed, 1 mol each of pyruvate and veratraldehyde is produced. The importance of these results is discussed in relation to the physiology of lignin biodegradation and possible extracellular regulatory mechanisms for the control of oxidase and peroxidase activities.</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">11607073</PMID><DateCompleted><Year>2001</Year><Month>12</Month><Day>20</Day></DateCompleted><DateRevised><Year>2019</Year><Month>05</Month><Day>01</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0027-8424</ISSN><JournalIssue CitedMedium="Print"><Volume>87</Volume><Issue>8</Issue><PubDate><Year>1990</Year><Month>Apr</Month></PubDate></JournalIssue><Title>Proceedings of the National Academy of Sciences of the United States of America</Title><ISOAbbreviation>Proc Natl Acad Sci U S A</ISOAbbreviation></Journal><ArticleTitle>Glyoxal oxidase of Phanerochaete chrysosporium: its characterization and activation by lignin peroxidase.</ArticleTitle><Pagination><MedlinePgn>2936-40</MedlinePgn></Pagination><Abstract><AbstractText>Glyoxal oxidase (GLOX) is an extracellular H2O2-generating enzyme produced by ligninolytic cultures of Phanerochaete chrysosporium. The production, purification, and partial characterization of GLOX from agitated cultures are described here. High-oxygen levels are critical for GLOX production as for lignin peroxidase. GLOX purified by anion-exchange chromatography appears homogeneous by NaDod-SO4/PAGE (molecular mass = 68 kDa). However, analysis by isoelectric focusing indicates two major bands (pI 4.7 and 4.9) that stain as glycoproteins as well as for H2O2-producing activity in the presence of methylglyoxal. Purified GLOX shows a marked stimulation in activity when incubated with Cu2+; full activation takes more than 1 hr with 1 mM CuSO4 at pH 6. The steady-state kinetic parameters for the GLOX oxidation of methylglyoxal, glyceraldehyde, dihydroxyacetone, glycolaldehyde, acetaldehyde, glyoxal, glyoxylic acid, and formaldehyde, were determined by using a lignin peroxidase coupled-assay at pH 4.5. Of these substrates, the best is the extracellular metabolite methylglyoxal with a Km of 0.64 mM an apparent rate of catalysis, kcat, of 198 s1 under air-saturated conditions. The Km for oxygen is greater than the concentration of oxygen possible at ambient pressure--i.e., >1.3 mM at 25 degrees C. Importantly, oxygen-uptake experiments show that purified GLOX is inactive unless coupled to the peroxidase reaction. With this coupled reaction, for each mol of methylglyoxal, veratryl alcohol (a lignin peroxidase substrate), and oxygen consumed, 1 mol each of pyruvate and veratraldehyde is produced. The importance of these results is discussed in relation to the physiology of lignin biodegradation and possible extracellular regulatory mechanisms for the control of oxidase and peroxidase activities.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Kersten</LastName><ForeName>P J</ForeName><Initials>PJ</Initials><AffiliationInfo><Affiliation>Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Proc Natl Acad Sci U S A</MedlineTA><NlmUniqueID>7505876</NlmUniqueID><ISSNLinking>0027-8424</ISSNLinking></MedlineJournalInfo></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1990</Year><Month>4</Month><Day>1</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2001</Year><Month>10</Month><Day>19</Day><Hour>10</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>1990</Year><Month>4</Month><Day>1</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">11607073</ArticleId><ArticleId IdType="pmc">PMC53808</ArticleId><ArticleId IdType="doi">10.1073/pnas.87.8.2936</ArticleId></ArticleIdList><ReferenceList><Reference><Citation>Proc Natl Acad Sci U S A. 1984 Apr;81(8):2280-4</Citation><ArticleIdList><ArticleId IdType="pubmed">16593451</ArticleId></ArticleIdList></Reference><Reference><Citation>Anal Biochem. 1976 May 7;72:248-54</Citation><ArticleIdList><ArticleId IdType="pubmed">942051</ArticleId></ArticleIdList></Reference><Reference><Citation>Science. 1983 Aug 12;221(4611):661-3</Citation><ArticleIdList><ArticleId IdType="pubmed">17787736</ArticleId></ArticleIdList></Reference><Reference><Citation>J Biol Chem. 1951 Jun;190(2):685-96</Citation><ArticleIdList><ArticleId IdType="pubmed">14841219</ArticleId></ArticleIdList></Reference><Reference><Citation>J Biol Chem. 1986 Feb 5;261(4):1687-93</Citation><ArticleIdList><ArticleId IdType="pubmed">3003081</ArticleId></ArticleIdList></Reference><Reference><Citation>Arch Biochem Biophys. 1985 Nov 1;242(2):329-41</Citation><ArticleIdList><ArticleId IdType="pubmed">4062285</ArticleId></ArticleIdList></Reference><Reference><Citation>J Biol Chem. 1986 May 25;261(15):6900-3</Citation><ArticleIdList><ArticleId IdType="pubmed">3700421</ArticleId></ArticleIdList></Reference><Reference><Citation>Annu Rev Microbiol. 1987;41:465-505</Citation><ArticleIdList><ArticleId IdType="pubmed">3318677</ArticleId></ArticleIdList></Reference><Reference><Citation>Crit Rev Microbiol. 1987;15(2):141-68</Citation><ArticleIdList><ArticleId IdType="pubmed">3322681</ArticleId></ArticleIdList></Reference><Reference><Citation>J Bacteriol. 1987 May;169(5):2195-201</Citation><ArticleIdList><ArticleId IdType="pubmed">3553159</ArticleId></ArticleIdList></Reference><Reference><Citation>J Biol Chem. 1988 May 5;263(13):6074-80</Citation><ArticleIdList><ArticleId IdType="pubmed">2834363</ArticleId></ArticleIdList></Reference><Reference><Citation>Arch Biochem Biophys. 1984 Nov 1;234(2):353-62</Citation><ArticleIdList><ArticleId IdType="pubmed">6497376</ArticleId></ArticleIdList></Reference><Reference><Citation>Biochem Biophys Res Commun. 1981 Mar 31;99(2):373-8</Citation><ArticleIdList><ArticleId IdType="pubmed">7236274</ArticleId></ArticleIdList></Reference><Reference><Citation>Biochem Biophys Res Commun. 1983 Aug 12;114(3):1077-83</Citation><ArticleIdList><ArticleId IdType="pubmed">6615503</ArticleId></ArticleIdList></Reference><Reference><Citation>Biochem Biophys Res Commun. 1984 Jan 30;118(2):437-43</Citation><ArticleIdList><ArticleId IdType="pubmed">6538411</ArticleId></ArticleIdList></Reference><Reference><Citation>Biochem Biophys Res Commun. 1983 Nov 30;117(1):275-81</Citation><ArticleIdList><ArticleId IdType="pubmed">6661224</ArticleId></ArticleIdList></Reference><Reference><Citation>Appl Environ Microbiol. 1985 Nov;50(5):1274-8</Citation><ArticleIdList><ArticleId IdType="pubmed">16346932</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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