Expression, and Molecular and Enzymatic Characterization of Cu-Containing Nitrite Reductase from a Marine Ammonia-Oxidizing Gammaproteobacterium, Nitrosococcus oceani
Ammonia-oxidizing bacteria (AOB) remove intracellular nitrite to prevent its toxicity by a nitrifier denitrification pathway involving two denitrifying enzymes, nitrite reductase and nitric oxide reductase. Here, a Cu-containing nitrite reductase from Nitrosococcus oceani strain NS58, a gammaproteob...
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| Veröffentlicht in: | Microbes and Environments 2012, Vol.27(4), pp.407-412 |
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| creator | Kondo, Keitaro Yoshimatsu, Katsuhiko Fujiwara, Taketomo |
| description | Ammonia-oxidizing bacteria (AOB) remove intracellular nitrite to prevent its toxicity by a nitrifier denitrification pathway involving two denitrifying enzymes, nitrite reductase and nitric oxide reductase. Here, a Cu-containing nitrite reductase from Nitrosococcus oceani strain NS58, a gammaproteobacterial marine AOB, was expressed in Escherichia coli and purified to homogeneity. Sequence homology analysis indicated that the nitrite reductase from N. oceani was phylogenetically closer to its counterparts from denitrifying bacteria than that of the betaproteobacterium Nitrosomonas europaea. The recombinant enzyme was a homotrimer of a 32 kDa subunit molecule. The enzyme was green in the oxidized state with absorption peaks at 455 nm and 575 nm. EPR spectroscopy indicated the presence of type 2 Cu. Molecular activities and the affinity constant for the nitrite were determined to be 1.6×103 s−1 and 52 mM, respectively. |
| doi_str_mv | 10.1264/jsme2.ME11310 |
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Here, a Cu-containing nitrite reductase from Nitrosococcus oceani strain NS58, a gammaproteobacterial marine AOB, was expressed in Escherichia coli and purified to homogeneity. Sequence homology analysis indicated that the nitrite reductase from N. oceani was phylogenetically closer to its counterparts from denitrifying bacteria than that of the betaproteobacterium Nitrosomonas europaea. The recombinant enzyme was a homotrimer of a 32 kDa subunit molecule. The enzyme was green in the oxidized state with absorption peaks at 455 nm and 575 nm. EPR spectroscopy indicated the presence of type 2 Cu. Molecular activities and the affinity constant for the nitrite were determined to be 1.6×103 s−1 and 52 mM, respectively.</description><identifier>ISSN: 1342-6311</identifier><identifier>EISSN: 1347-4405</identifier><identifier>DOI: 10.1264/jsme2.ME11310</identifier><identifier>PMID: 22641151</identifier><language>eng</language><publisher>Japan: Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology / Japanese Society of Plant Microbe Interactions / Japanese Society for Extremophiles</publisher><subject>Ammonia - metabolism ; Base Sequence ; Cu-containing nitrite reductase ; Denitrification ; Escherichia coli ; Escherichia coli - genetics ; Gammaproteobacteria - enzymology ; Gammaproteobacteria - genetics ; Gammaproteobacteria - metabolism ; marine ammonia-oxidizing gammaproteobacteria ; Molecular Sequence Data ; nitrifier denitrification pathway ; Nitrite Reductases - chemistry ; Nitrite Reductases - genetics ; Nitrite Reductases - metabolism ; Nitrosococcus oceani ; Nitrosomonas europaea ; Nitrosomonas europaea - enzymology ; Oxidation-Reduction ; Phylogeny ; Recombinant Proteins - biosynthesis ; Sequence Analysis, DNA</subject><ispartof>Microbes and Environments, 2012, Vol.27(4), pp.407-412</ispartof><rights>2012 Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology</rights><rights>Copyright Japan Science and Technology Agency 2012</rights><rights>Copyright © 2012 by the Japanese Society of Microbial Ecology / the Japanese Society of Soil Microbiology 2012</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c625t-bb2d3a488ee583004834dc6b89351c0f785034911804142a6ba601aac2b9f4483</citedby><cites>FETCH-LOGICAL-c625t-bb2d3a488ee583004834dc6b89351c0f785034911804142a6ba601aac2b9f4483</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4103548/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4103548/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,724,777,781,882,1877,4010,27904,27905,27906,53772,53774</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22641151$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kondo, Keitaro</creatorcontrib><creatorcontrib>Yoshimatsu, Katsuhiko</creatorcontrib><creatorcontrib>Fujiwara, Taketomo</creatorcontrib><title>Expression, and Molecular and Enzymatic Characterization of Cu-Containing Nitrite Reductase from a Marine Ammonia-Oxidizing Gammaproteobacterium, Nitrosococcus oceani</title><title>Microbes and Environments</title><addtitle>Microbes Environ.</addtitle><description>Ammonia-oxidizing bacteria (AOB) remove intracellular nitrite to prevent its toxicity by a nitrifier denitrification pathway involving two denitrifying enzymes, nitrite reductase and nitric oxide reductase. Here, a Cu-containing nitrite reductase from Nitrosococcus oceani strain NS58, a gammaproteobacterial marine AOB, was expressed in Escherichia coli and purified to homogeneity. Sequence homology analysis indicated that the nitrite reductase from N. oceani was phylogenetically closer to its counterparts from denitrifying bacteria than that of the betaproteobacterium Nitrosomonas europaea. The recombinant enzyme was a homotrimer of a 32 kDa subunit molecule. The enzyme was green in the oxidized state with absorption peaks at 455 nm and 575 nm. EPR spectroscopy indicated the presence of type 2 Cu. Molecular activities and the affinity constant for the nitrite were determined to be 1.6×103 s−1 and 52 mM, respectively.</description><subject>Ammonia - metabolism</subject><subject>Base Sequence</subject><subject>Cu-containing nitrite reductase</subject><subject>Denitrification</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Gammaproteobacteria - enzymology</subject><subject>Gammaproteobacteria - genetics</subject><subject>Gammaproteobacteria - metabolism</subject><subject>marine ammonia-oxidizing gammaproteobacteria</subject><subject>Molecular Sequence Data</subject><subject>nitrifier denitrification pathway</subject><subject>Nitrite Reductases - chemistry</subject><subject>Nitrite Reductases - genetics</subject><subject>Nitrite Reductases - metabolism</subject><subject>Nitrosococcus oceani</subject><subject>Nitrosomonas europaea</subject><subject>Nitrosomonas europaea - enzymology</subject><subject>Oxidation-Reduction</subject><subject>Phylogeny</subject><subject>Recombinant Proteins - biosynthesis</subject><subject>Sequence Analysis, DNA</subject><issn>1342-6311</issn><issn>1347-4405</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkk1v1DAQhiMEoqVw5IosceHQFDt2PvaCVEVLQepSCcHZmjiTrVexvdgOavcH8Tvx7oYVcLE9mmdezXjeLHvN6BUrKvF-EwwWV6slY5zRJ9k546LOhaDl08O7yCvO2Fn2IoQNpZyXdfE8OytSJWMlO89-LR-2HkPQzl4SsD1ZuRHVNII_REu7ezQQtSLtPXhQEb3epdhZ4gbSTnnrbARttV2TLzp6HZF8xX5SEQKSwTtDgKzAa4vk2hhnNeR3D7rXu33FDRgDW-8iuu6oPZnLg44LTjmlpkCcQrD6ZfZsgDHgq_m-yL5_XH5rP-W3dzef2-vbXFVFGfOuK3oOomkQy4ZTKhouelV1zYKXTNGhbkrKxYKxhgomCqg6qCgDUEW3GESiL7IPR93t1BnsFdroYZRbrw34R-lAy38zVt_LtfspBaO8PAi8mwW8-zFhiNLooHAcwaKbgmS8qhPW0EVC3_6HbtzkbRpPMlHVtajook5UfqRU-pTgcTg1w6jcO0AeHCBnByT-zd8TnOg_K09AewQ2IcIaTwD4tOYRZ7milmJ_zLKnrEo2kGj5b12PyEg</recordid><startdate>2012</startdate><enddate>2012</enddate><creator>Kondo, Keitaro</creator><creator>Yoshimatsu, Katsuhiko</creator><creator>Fujiwara, Taketomo</creator><general>Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology / Japanese Society of Plant Microbe Interactions / Japanese Society for Extremophiles</general><general>Japan Science and Technology Agency</general><general>Japanese Society of Microbial Ecology/The Japanese Society of Soil Microbiology</general><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>7QL</scope><scope>C1K</scope><scope>F1W</scope><scope>H95</scope><scope>L.G</scope><scope>M7N</scope><scope>7TN</scope><scope>5PM</scope></search><sort><creationdate>2012</creationdate><title>Expression, and Molecular and Enzymatic Characterization of Cu-Containing Nitrite Reductase from a Marine Ammonia-Oxidizing Gammaproteobacterium, Nitrosococcus oceani</title><author>Kondo, Keitaro ; Yoshimatsu, Katsuhiko ; Fujiwara, Taketomo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c625t-bb2d3a488ee583004834dc6b89351c0f785034911804142a6ba601aac2b9f4483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Ammonia - metabolism</topic><topic>Base Sequence</topic><topic>Cu-containing nitrite reductase</topic><topic>Denitrification</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Gammaproteobacteria - enzymology</topic><topic>Gammaproteobacteria - genetics</topic><topic>Gammaproteobacteria - metabolism</topic><topic>marine ammonia-oxidizing gammaproteobacteria</topic><topic>Molecular Sequence Data</topic><topic>nitrifier denitrification pathway</topic><topic>Nitrite Reductases - chemistry</topic><topic>Nitrite Reductases - genetics</topic><topic>Nitrite Reductases - metabolism</topic><topic>Nitrosococcus oceani</topic><topic>Nitrosomonas europaea</topic><topic>Nitrosomonas europaea - enzymology</topic><topic>Oxidation-Reduction</topic><topic>Phylogeny</topic><topic>Recombinant Proteins - biosynthesis</topic><topic>Sequence Analysis, DNA</topic><toplevel>online_resources</toplevel><creatorcontrib>Kondo, Keitaro</creatorcontrib><creatorcontrib>Yoshimatsu, Katsuhiko</creatorcontrib><creatorcontrib>Fujiwara, Taketomo</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Oceanic Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Microbes and Environments</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kondo, Keitaro</au><au>Yoshimatsu, Katsuhiko</au><au>Fujiwara, Taketomo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression, and Molecular and Enzymatic Characterization of Cu-Containing Nitrite Reductase from a Marine Ammonia-Oxidizing Gammaproteobacterium, Nitrosococcus oceani</atitle><jtitle>Microbes and Environments</jtitle><addtitle>Microbes Environ.</addtitle><date>2012</date><risdate>2012</risdate><volume>27</volume><issue>4</issue><spage>407</spage><epage>412</epage><pages>407-412</pages><issn>1342-6311</issn><eissn>1347-4405</eissn><abstract>Ammonia-oxidizing bacteria (AOB) remove intracellular nitrite to prevent its toxicity by a nitrifier denitrification pathway involving two denitrifying enzymes, nitrite reductase and nitric oxide reductase. Here, a Cu-containing nitrite reductase from Nitrosococcus oceani strain NS58, a gammaproteobacterial marine AOB, was expressed in Escherichia coli and purified to homogeneity. Sequence homology analysis indicated that the nitrite reductase from N. oceani was phylogenetically closer to its counterparts from denitrifying bacteria than that of the betaproteobacterium Nitrosomonas europaea. The recombinant enzyme was a homotrimer of a 32 kDa subunit molecule. The enzyme was green in the oxidized state with absorption peaks at 455 nm and 575 nm. EPR spectroscopy indicated the presence of type 2 Cu. 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| subjects | Ammonia - metabolism Base Sequence Cu-containing nitrite reductase Denitrification Escherichia coli Escherichia coli - genetics Gammaproteobacteria - enzymology Gammaproteobacteria - genetics Gammaproteobacteria - metabolism marine ammonia-oxidizing gammaproteobacteria Molecular Sequence Data nitrifier denitrification pathway Nitrite Reductases - chemistry Nitrite Reductases - genetics Nitrite Reductases - metabolism Nitrosococcus oceani Nitrosomonas europaea Nitrosomonas europaea - enzymology Oxidation-Reduction Phylogeny Recombinant Proteins - biosynthesis Sequence Analysis, DNA |
| title | Expression, and Molecular and Enzymatic Characterization of Cu-Containing Nitrite Reductase from a Marine Ammonia-Oxidizing Gammaproteobacterium, Nitrosococcus oceani |
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