Characterization of a catalase-peroxidase from the hyperthermophilic archaeon Archaeoglobus fulgidus
A putative perA gene from Archaeoglobus fulgidus was cloned and expressed in Escherichia coli BL21(DE3), and the recombinant catalase-peroxidase was purified to homogeneity. The enzyme is a homodimer with a subunit molecular mass of 85 kDa. UV-visible spectroscopic analysis indicated the presence of...
Gespeichert in:
Veröffentlicht in: | Extremophiles : life under extreme conditions 2001-10, Vol.5 (5), p.323-332 |
---|---|
Hauptverfasser: | , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 332 |
---|---|
container_issue | 5 |
container_start_page | 323 |
container_title | Extremophiles : life under extreme conditions |
container_volume | 5 |
creator | Kengen, S W Bikker, F J Hagen, W R de Vos, W M van der Oost, J |
description | A putative perA gene from Archaeoglobus fulgidus was cloned and expressed in Escherichia coli BL21(DE3), and the recombinant catalase-peroxidase was purified to homogeneity. The enzyme is a homodimer with a subunit molecular mass of 85 kDa. UV-visible spectroscopic analysis indicated the presence of protoheme IX as a prosthetic group (ferric heme), in a stoichiometry of 0.25 heme per subunit. Electron paramagnetic resonance analysis confirmed the presence of ferric heme and identified the proximal axial ligand as a histidine. The enzyme showed both catalase and peroxidase activity with pH optima of 6.0 and 4.5, respectively. Optimal temperatures of 70 degrees C and 80 degrees C were found for the catalase and peroxidase activity, respectively. The catalase activity strongly exceeded the peroxidase activity, with Vmax values of 9600 and 36 U mg(-1), respectively. Km values for H2O2 of 8.6 and 0.85 mM were found for catalase and peroxidase, respectively. Common heme inhibitors such as cyanide, azide, and hydroxylamine inhibited peroxidase activity. However, unlike all other catalase-peroxidases, the enzyme was also inhibited by 3-amino-1,2,4-triazole. Although the enzyme exhibited a high thermostability, rapid inactivation occurred in the presence of H2O2, with half-life values of less than 1 min. This is the first catalase-peroxidase characterized from a hyperthermophilic microorganism. |
doi_str_mv | 10.1007/s007920100208 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_72267389</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>72267389</sourcerecordid><originalsourceid>FETCH-LOGICAL-c355t-6d58285acb18decd1d394864bded629f7fef73dbb1151af90545b8efa93781513</originalsourceid><addsrcrecordid>eNpVkL1PwzAQxS0EolAYWZEntoAdx449VhVfUiUWmKOLPxqjpC52IlH-egythFjufnr37g0PoStKbikh9V3KQ5Ukc0nkETqjFWNFpYg6_mVaEMHpDJ2n9E4I5flwimaUCqVEJc6QWXYQQY82-i8Yfdjg4DBgDSP0kGyxtTF8epMRuxgGPHYWd7usZohD2Ha-9xpD1B3Y_LzYw7oP7ZSwm_q1N1O6QCcO-mQvD3uO3h7uX5dPxerl8Xm5WBWacT4WwnBZSg66pdJYbahhqpKiao01olSudtbVzLQtpZyCU4RXvJXWgWK1zBKbo5t97jaGj8mmsRl80rbvYWPDlJq6LEXNpMrGYm_UMaQUrWu20Q8Qdw0lzU-tzb9as__6EDy1gzV_7kOP7BtLv3TM</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>72267389</pqid></control><display><type>article</type><title>Characterization of a catalase-peroxidase from the hyperthermophilic archaeon Archaeoglobus fulgidus</title><source>MEDLINE</source><source>SpringerLink Journals - AutoHoldings</source><creator>Kengen, S W ; Bikker, F J ; Hagen, W R ; de Vos, W M ; van der Oost, J</creator><creatorcontrib>Kengen, S W ; Bikker, F J ; Hagen, W R ; de Vos, W M ; van der Oost, J</creatorcontrib><description>A putative perA gene from Archaeoglobus fulgidus was cloned and expressed in Escherichia coli BL21(DE3), and the recombinant catalase-peroxidase was purified to homogeneity. The enzyme is a homodimer with a subunit molecular mass of 85 kDa. UV-visible spectroscopic analysis indicated the presence of protoheme IX as a prosthetic group (ferric heme), in a stoichiometry of 0.25 heme per subunit. Electron paramagnetic resonance analysis confirmed the presence of ferric heme and identified the proximal axial ligand as a histidine. The enzyme showed both catalase and peroxidase activity with pH optima of 6.0 and 4.5, respectively. Optimal temperatures of 70 degrees C and 80 degrees C were found for the catalase and peroxidase activity, respectively. The catalase activity strongly exceeded the peroxidase activity, with Vmax values of 9600 and 36 U mg(-1), respectively. Km values for H2O2 of 8.6 and 0.85 mM were found for catalase and peroxidase, respectively. Common heme inhibitors such as cyanide, azide, and hydroxylamine inhibited peroxidase activity. However, unlike all other catalase-peroxidases, the enzyme was also inhibited by 3-amino-1,2,4-triazole. Although the enzyme exhibited a high thermostability, rapid inactivation occurred in the presence of H2O2, with half-life values of less than 1 min. This is the first catalase-peroxidase characterized from a hyperthermophilic microorganism.</description><identifier>ISSN: 1431-0651</identifier><identifier>EISSN: 1433-4909</identifier><identifier>DOI: 10.1007/s007920100208</identifier><identifier>PMID: 11699646</identifier><language>eng</language><publisher>Germany</publisher><subject>Archaeoglobus fulgidus - enzymology ; Archaeoglobus fulgidus - genetics ; Catalase - antagonists & inhibitors ; Catalase - chemistry ; Catalase - genetics ; Catalase - metabolism ; Cloning, Molecular ; Electron Spin Resonance Spectroscopy ; Enzyme Inhibitors - pharmacology ; Genes, Archaeal ; Hot Temperature ; Kinetics ; Peroxidases - antagonists & inhibitors ; Peroxidases - chemistry ; Peroxidases - genetics ; Peroxidases - metabolism ; Phylogeny ; Recombinant Proteins - chemistry ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Sequence Analysis, Protein ; Space life sciences ; Spectrophotometry</subject><ispartof>Extremophiles : life under extreme conditions, 2001-10, Vol.5 (5), p.323-332</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c355t-6d58285acb18decd1d394864bded629f7fef73dbb1151af90545b8efa93781513</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11699646$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kengen, S W</creatorcontrib><creatorcontrib>Bikker, F J</creatorcontrib><creatorcontrib>Hagen, W R</creatorcontrib><creatorcontrib>de Vos, W M</creatorcontrib><creatorcontrib>van der Oost, J</creatorcontrib><title>Characterization of a catalase-peroxidase from the hyperthermophilic archaeon Archaeoglobus fulgidus</title><title>Extremophiles : life under extreme conditions</title><addtitle>Extremophiles</addtitle><description>A putative perA gene from Archaeoglobus fulgidus was cloned and expressed in Escherichia coli BL21(DE3), and the recombinant catalase-peroxidase was purified to homogeneity. The enzyme is a homodimer with a subunit molecular mass of 85 kDa. UV-visible spectroscopic analysis indicated the presence of protoheme IX as a prosthetic group (ferric heme), in a stoichiometry of 0.25 heme per subunit. Electron paramagnetic resonance analysis confirmed the presence of ferric heme and identified the proximal axial ligand as a histidine. The enzyme showed both catalase and peroxidase activity with pH optima of 6.0 and 4.5, respectively. Optimal temperatures of 70 degrees C and 80 degrees C were found for the catalase and peroxidase activity, respectively. The catalase activity strongly exceeded the peroxidase activity, with Vmax values of 9600 and 36 U mg(-1), respectively. Km values for H2O2 of 8.6 and 0.85 mM were found for catalase and peroxidase, respectively. Common heme inhibitors such as cyanide, azide, and hydroxylamine inhibited peroxidase activity. However, unlike all other catalase-peroxidases, the enzyme was also inhibited by 3-amino-1,2,4-triazole. Although the enzyme exhibited a high thermostability, rapid inactivation occurred in the presence of H2O2, with half-life values of less than 1 min. This is the first catalase-peroxidase characterized from a hyperthermophilic microorganism.</description><subject>Archaeoglobus fulgidus - enzymology</subject><subject>Archaeoglobus fulgidus - genetics</subject><subject>Catalase - antagonists & inhibitors</subject><subject>Catalase - chemistry</subject><subject>Catalase - genetics</subject><subject>Catalase - metabolism</subject><subject>Cloning, Molecular</subject><subject>Electron Spin Resonance Spectroscopy</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Genes, Archaeal</subject><subject>Hot Temperature</subject><subject>Kinetics</subject><subject>Peroxidases - antagonists & inhibitors</subject><subject>Peroxidases - chemistry</subject><subject>Peroxidases - genetics</subject><subject>Peroxidases - metabolism</subject><subject>Phylogeny</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>Sequence Analysis, Protein</subject><subject>Space life sciences</subject><subject>Spectrophotometry</subject><issn>1431-0651</issn><issn>1433-4909</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkL1PwzAQxS0EolAYWZEntoAdx449VhVfUiUWmKOLPxqjpC52IlH-egythFjufnr37g0PoStKbikh9V3KQ5Ukc0nkETqjFWNFpYg6_mVaEMHpDJ2n9E4I5flwimaUCqVEJc6QWXYQQY82-i8Yfdjg4DBgDSP0kGyxtTF8epMRuxgGPHYWd7usZohD2Ha-9xpD1B3Y_LzYw7oP7ZSwm_q1N1O6QCcO-mQvD3uO3h7uX5dPxerl8Xm5WBWacT4WwnBZSg66pdJYbahhqpKiao01olSudtbVzLQtpZyCU4RXvJXWgWK1zBKbo5t97jaGj8mmsRl80rbvYWPDlJq6LEXNpMrGYm_UMaQUrWu20Q8Qdw0lzU-tzb9as__6EDy1gzV_7kOP7BtLv3TM</recordid><startdate>20011001</startdate><enddate>20011001</enddate><creator>Kengen, S W</creator><creator>Bikker, F J</creator><creator>Hagen, W R</creator><creator>de Vos, W M</creator><creator>van der Oost, J</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20011001</creationdate><title>Characterization of a catalase-peroxidase from the hyperthermophilic archaeon Archaeoglobus fulgidus</title><author>Kengen, S W ; Bikker, F J ; Hagen, W R ; de Vos, W M ; van der Oost, J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c355t-6d58285acb18decd1d394864bded629f7fef73dbb1151af90545b8efa93781513</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Archaeoglobus fulgidus - enzymology</topic><topic>Archaeoglobus fulgidus - genetics</topic><topic>Catalase - antagonists & inhibitors</topic><topic>Catalase - chemistry</topic><topic>Catalase - genetics</topic><topic>Catalase - metabolism</topic><topic>Cloning, Molecular</topic><topic>Electron Spin Resonance Spectroscopy</topic><topic>Enzyme Inhibitors - pharmacology</topic><topic>Genes, Archaeal</topic><topic>Hot Temperature</topic><topic>Kinetics</topic><topic>Peroxidases - antagonists & inhibitors</topic><topic>Peroxidases - chemistry</topic><topic>Peroxidases - genetics</topic><topic>Peroxidases - metabolism</topic><topic>Phylogeny</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><topic>Sequence Analysis, Protein</topic><topic>Space life sciences</topic><topic>Spectrophotometry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kengen, S W</creatorcontrib><creatorcontrib>Bikker, F J</creatorcontrib><creatorcontrib>Hagen, W R</creatorcontrib><creatorcontrib>de Vos, W M</creatorcontrib><creatorcontrib>van der Oost, J</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Extremophiles : life under extreme conditions</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kengen, S W</au><au>Bikker, F J</au><au>Hagen, W R</au><au>de Vos, W M</au><au>van der Oost, J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of a catalase-peroxidase from the hyperthermophilic archaeon Archaeoglobus fulgidus</atitle><jtitle>Extremophiles : life under extreme conditions</jtitle><addtitle>Extremophiles</addtitle><date>2001-10-01</date><risdate>2001</risdate><volume>5</volume><issue>5</issue><spage>323</spage><epage>332</epage><pages>323-332</pages><issn>1431-0651</issn><eissn>1433-4909</eissn><abstract>A putative perA gene from Archaeoglobus fulgidus was cloned and expressed in Escherichia coli BL21(DE3), and the recombinant catalase-peroxidase was purified to homogeneity. The enzyme is a homodimer with a subunit molecular mass of 85 kDa. UV-visible spectroscopic analysis indicated the presence of protoheme IX as a prosthetic group (ferric heme), in a stoichiometry of 0.25 heme per subunit. Electron paramagnetic resonance analysis confirmed the presence of ferric heme and identified the proximal axial ligand as a histidine. The enzyme showed both catalase and peroxidase activity with pH optima of 6.0 and 4.5, respectively. Optimal temperatures of 70 degrees C and 80 degrees C were found for the catalase and peroxidase activity, respectively. The catalase activity strongly exceeded the peroxidase activity, with Vmax values of 9600 and 36 U mg(-1), respectively. Km values for H2O2 of 8.6 and 0.85 mM were found for catalase and peroxidase, respectively. Common heme inhibitors such as cyanide, azide, and hydroxylamine inhibited peroxidase activity. However, unlike all other catalase-peroxidases, the enzyme was also inhibited by 3-amino-1,2,4-triazole. Although the enzyme exhibited a high thermostability, rapid inactivation occurred in the presence of H2O2, with half-life values of less than 1 min. This is the first catalase-peroxidase characterized from a hyperthermophilic microorganism.</abstract><cop>Germany</cop><pmid>11699646</pmid><doi>10.1007/s007920100208</doi><tpages>10</tpages></addata></record> |
fulltext | fulltext |
identifier | ISSN: 1431-0651 |
ispartof | Extremophiles : life under extreme conditions, 2001-10, Vol.5 (5), p.323-332 |
issn | 1431-0651 1433-4909 |
language | eng |
recordid | cdi_proquest_miscellaneous_72267389 |
source | MEDLINE; SpringerLink Journals - AutoHoldings |
subjects | Archaeoglobus fulgidus - enzymology Archaeoglobus fulgidus - genetics Catalase - antagonists & inhibitors Catalase - chemistry Catalase - genetics Catalase - metabolism Cloning, Molecular Electron Spin Resonance Spectroscopy Enzyme Inhibitors - pharmacology Genes, Archaeal Hot Temperature Kinetics Peroxidases - antagonists & inhibitors Peroxidases - chemistry Peroxidases - genetics Peroxidases - metabolism Phylogeny Recombinant Proteins - chemistry Recombinant Proteins - genetics Recombinant Proteins - metabolism Sequence Analysis, Protein Space life sciences Spectrophotometry |
title | Characterization of a catalase-peroxidase from the hyperthermophilic archaeon Archaeoglobus fulgidus |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-12T01%3A07%3A27IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Characterization%20of%20a%20catalase-peroxidase%20from%20the%20hyperthermophilic%20archaeon%20Archaeoglobus%20fulgidus&rft.jtitle=Extremophiles%20:%20life%20under%20extreme%20conditions&rft.au=Kengen,%20S%20W&rft.date=2001-10-01&rft.volume=5&rft.issue=5&rft.spage=323&rft.epage=332&rft.pages=323-332&rft.issn=1431-0651&rft.eissn=1433-4909&rft_id=info:doi/10.1007/s007920100208&rft_dat=%3Cproquest_cross%3E72267389%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=72267389&rft_id=info:pmid/11699646&rfr_iscdi=true |