Heterologous expression of heterotrophic nitrification genes
School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK Author for correspondence: Stephen Spiro. Tel: + 44 1603 592777. Fax: +44 1603 592250. e-mail: s.spiro@uea.ac.uk ABSTRACT Paracoccus denitrificans is a heterotrophic organism capable of oxidizing ammonia to nitrite during...
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| Veröffentlicht in: | Microbiology (Society for General Microbiology) 1997-12, Vol.143 (12), p.3775-3783 |
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| creator | Crossman, Lisa C Moir, James W. B Enticknap, Julie J Richardson, David J Spiro, Stephen |
| description | School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK
Author for correspondence: Stephen Spiro. Tel: + 44 1603 592777. Fax: +44 1603 592250. e-mail: s.spiro@uea.ac.uk
ABSTRACT
Paracoccus denitrificans is a heterotrophic organism capable of oxidizing ammonia to nitrite during growth on an organic carbon and energy source. This pathway, termed heterotrophic nitrification, requires the concerted action of an ammonia monooxygenase (AMO) and hydroxylamine oxidase (HAO). The genes required for heterotrophic nitrification have been isolated by introducing a Pa. denitrificans genomic library into Pseudomonas putida and screening for the accumulation of nitrite. In contrast to the situation in chemolithoautotrophic ammonia oxidizers, the genes encoding AMO and HAO are present in single linked copies in the genome of Pa. denitrificans. AMO from Pa. denitrificans expressed in Ps. putida is capable of oxidizing ethene (ethylene) to epoxyethane (ethylene oxide), which is indicative of a relaxed substrate specificity. Further, when expressed in the methylotroph Methylobacterium extorquens AM1, the AMO endows on this organism the ability to grow on ethene and methane. Thus, the Pa. denitrificans AMO is capable of oxidizing methane to methanol, as is the case for the AMO from Nitrosomonas europaea. The heterotrophic nitrification genes are moderately toxic in M. extorquens, more toxic in Ps. putida, and non-toxic in Escherichia coli. Toxicity is due to the activity of the gene products in M. extorquens, and both expression and activity in Ps. putida. This is the first time that the genes encoding an active AMO have been expressed in a heterologous host.
Keywords: heterotrophic nitrification, ammonia monooxygenase, hydroxylamine oxidoreductase, Paracoccus denitrificans, Methylobacterium extorquens
Present address: Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.
Present address: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, UK. |
| doi_str_mv | 10.1099/00221287-143-12-3775 |
| format | Article |
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Author for correspondence: Stephen Spiro. Tel: + 44 1603 592777. Fax: +44 1603 592250. e-mail: s.spiro@uea.ac.uk
ABSTRACT
Paracoccus denitrificans is a heterotrophic organism capable of oxidizing ammonia to nitrite during growth on an organic carbon and energy source. This pathway, termed heterotrophic nitrification, requires the concerted action of an ammonia monooxygenase (AMO) and hydroxylamine oxidase (HAO). The genes required for heterotrophic nitrification have been isolated by introducing a Pa. denitrificans genomic library into Pseudomonas putida and screening for the accumulation of nitrite. In contrast to the situation in chemolithoautotrophic ammonia oxidizers, the genes encoding AMO and HAO are present in single linked copies in the genome of Pa. denitrificans. AMO from Pa. denitrificans expressed in Ps. putida is capable of oxidizing ethene (ethylene) to epoxyethane (ethylene oxide), which is indicative of a relaxed substrate specificity. Further, when expressed in the methylotroph Methylobacterium extorquens AM1, the AMO endows on this organism the ability to grow on ethene and methane. Thus, the Pa. denitrificans AMO is capable of oxidizing methane to methanol, as is the case for the AMO from Nitrosomonas europaea. The heterotrophic nitrification genes are moderately toxic in M. extorquens, more toxic in Ps. putida, and non-toxic in Escherichia coli. Toxicity is due to the activity of the gene products in M. extorquens, and both expression and activity in Ps. putida. This is the first time that the genes encoding an active AMO have been expressed in a heterologous host.
Keywords: heterotrophic nitrification, ammonia monooxygenase, hydroxylamine oxidoreductase, Paracoccus denitrificans, Methylobacterium extorquens
Present address: Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.
Present address: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, UK.</description><identifier>ISSN: 1350-0872</identifier><identifier>EISSN: 1465-2080</identifier><identifier>DOI: 10.1099/00221287-143-12-3775</identifier><identifier>PMID: 9421902</identifier><language>eng</language><publisher>Reading: Soc General Microbiol</publisher><subject>Ammonia - metabolism ; Bacteriology ; Biological and medical sciences ; Cloning, Molecular - methods ; Cosmids ; Fundamental and applied biological sciences. Psychology ; Genes, Bacterial ; Genetics ; Genomic Library ; Methylobacterium extorquens ; Microbiology ; Nitrate Reductase ; Nitrate Reductases - metabolism ; Nitrites - metabolism ; Oxidoreductases - genetics ; Oxidoreductases - metabolism ; Paracoccus denitrificans ; Paracoccus denitrificans - genetics ; Paracoccus denitrificans - growth & development ; Paracoccus denitrificans - metabolism ; Pseudomonas putida - growth & development ; Pseudomonas putida - metabolism ; Recombinant Proteins - metabolism ; Restriction Mapping</subject><ispartof>Microbiology (Society for General Microbiology), 1997-12, Vol.143 (12), p.3775-3783</ispartof><rights>1998 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c546t-be5d26f62bad222b32638c3a4280995f9bfff38aec2028033498845c7c2522943</citedby><cites>FETCH-LOGICAL-c546t-be5d26f62bad222b32638c3a4280995f9bfff38aec2028033498845c7c2522943</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2144418$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9421902$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Crossman, Lisa C</creatorcontrib><creatorcontrib>Moir, James W. B</creatorcontrib><creatorcontrib>Enticknap, Julie J</creatorcontrib><creatorcontrib>Richardson, David J</creatorcontrib><creatorcontrib>Spiro, Stephen</creatorcontrib><title>Heterologous expression of heterotrophic nitrification genes</title><title>Microbiology (Society for General Microbiology)</title><addtitle>Microbiology (Reading)</addtitle><description>School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK
Author for correspondence: Stephen Spiro. Tel: + 44 1603 592777. Fax: +44 1603 592250. e-mail: s.spiro@uea.ac.uk
ABSTRACT
Paracoccus denitrificans is a heterotrophic organism capable of oxidizing ammonia to nitrite during growth on an organic carbon and energy source. This pathway, termed heterotrophic nitrification, requires the concerted action of an ammonia monooxygenase (AMO) and hydroxylamine oxidase (HAO). The genes required for heterotrophic nitrification have been isolated by introducing a Pa. denitrificans genomic library into Pseudomonas putida and screening for the accumulation of nitrite. In contrast to the situation in chemolithoautotrophic ammonia oxidizers, the genes encoding AMO and HAO are present in single linked copies in the genome of Pa. denitrificans. AMO from Pa. denitrificans expressed in Ps. putida is capable of oxidizing ethene (ethylene) to epoxyethane (ethylene oxide), which is indicative of a relaxed substrate specificity. Further, when expressed in the methylotroph Methylobacterium extorquens AM1, the AMO endows on this organism the ability to grow on ethene and methane. Thus, the Pa. denitrificans AMO is capable of oxidizing methane to methanol, as is the case for the AMO from Nitrosomonas europaea. The heterotrophic nitrification genes are moderately toxic in M. extorquens, more toxic in Ps. putida, and non-toxic in Escherichia coli. Toxicity is due to the activity of the gene products in M. extorquens, and both expression and activity in Ps. putida. This is the first time that the genes encoding an active AMO have been expressed in a heterologous host.
Keywords: heterotrophic nitrification, ammonia monooxygenase, hydroxylamine oxidoreductase, Paracoccus denitrificans, Methylobacterium extorquens
Present address: Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.
Present address: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, UK.</description><subject>Ammonia - metabolism</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Cloning, Molecular - methods</subject><subject>Cosmids</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes, Bacterial</subject><subject>Genetics</subject><subject>Genomic Library</subject><subject>Methylobacterium extorquens</subject><subject>Microbiology</subject><subject>Nitrate Reductase</subject><subject>Nitrate Reductases - metabolism</subject><subject>Nitrites - metabolism</subject><subject>Oxidoreductases - genetics</subject><subject>Oxidoreductases - metabolism</subject><subject>Paracoccus denitrificans</subject><subject>Paracoccus denitrificans - genetics</subject><subject>Paracoccus denitrificans - growth & development</subject><subject>Paracoccus denitrificans - metabolism</subject><subject>Pseudomonas putida - growth & development</subject><subject>Pseudomonas putida - metabolism</subject><subject>Recombinant Proteins - metabolism</subject><subject>Restriction Mapping</subject><issn>1350-0872</issn><issn>1465-2080</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpNkEFLxDAQhYMo67r6DxR6EA9CNZmkbQpeZFFXWPCi55Bmk22kbWrSRf33Zt26eJow783My4fQOcE3BJflLcYABHiREkZTAiktiuwATQnLsxQwx4fxTTOcYl7AMToJ4R3jKGIyQZOSASkxTNHdQg_au8at3SYk-qv3OgTrusSZpP6VBu_62qqks4O3xio5bOW17nQ4RUdGNkGfjXWG3h4fXueLdPny9Dy_X6YqY_mQVjpbQW5yqOQKACoKOeWKSgY8_iMzZWWMoVxqBTi2KGUl5yxThYIMoGR0hq52e3vvPjY6DKK1QemmkZ2OsQUpCC44gWhkO6PyLgSvjei9baX_FgSLLTTxB01EaIKA2EKLYxfj_k3V6tV-aKQU9ctRl0HJxnjZKRv2NiCMMcKj7Xpnq-26_rRei0iptTFLZV2MrP7f_AGanYH3</recordid><startdate>19971201</startdate><enddate>19971201</enddate><creator>Crossman, Lisa C</creator><creator>Moir, James W. B</creator><creator>Enticknap, Julie J</creator><creator>Richardson, David J</creator><creator>Spiro, Stephen</creator><general>Soc General Microbiol</general><general>Society for General Microbiology</general><scope>IQODW</scope><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>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>19971201</creationdate><title>Heterologous expression of heterotrophic nitrification genes</title><author>Crossman, Lisa C ; Moir, James W. B ; Enticknap, Julie J ; Richardson, David J ; Spiro, Stephen</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c546t-be5d26f62bad222b32638c3a4280995f9bfff38aec2028033498845c7c2522943</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Ammonia - metabolism</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Cloning, Molecular - methods</topic><topic>Cosmids</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes, Bacterial</topic><topic>Genetics</topic><topic>Genomic Library</topic><topic>Methylobacterium extorquens</topic><topic>Microbiology</topic><topic>Nitrate Reductase</topic><topic>Nitrate Reductases - metabolism</topic><topic>Nitrites - metabolism</topic><topic>Oxidoreductases - genetics</topic><topic>Oxidoreductases - metabolism</topic><topic>Paracoccus denitrificans</topic><topic>Paracoccus denitrificans - genetics</topic><topic>Paracoccus denitrificans - growth & development</topic><topic>Paracoccus denitrificans - metabolism</topic><topic>Pseudomonas putida - growth & development</topic><topic>Pseudomonas putida - metabolism</topic><topic>Recombinant Proteins - metabolism</topic><topic>Restriction Mapping</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Crossman, Lisa C</creatorcontrib><creatorcontrib>Moir, James W. B</creatorcontrib><creatorcontrib>Enticknap, Julie J</creatorcontrib><creatorcontrib>Richardson, David J</creatorcontrib><creatorcontrib>Spiro, Stephen</creatorcontrib><collection>Pascal-Francis</collection><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>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Microbiology (Society for General Microbiology)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Crossman, Lisa C</au><au>Moir, James W. B</au><au>Enticknap, Julie J</au><au>Richardson, David J</au><au>Spiro, Stephen</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Heterologous expression of heterotrophic nitrification genes</atitle><jtitle>Microbiology (Society for General Microbiology)</jtitle><addtitle>Microbiology (Reading)</addtitle><date>1997-12-01</date><risdate>1997</risdate><volume>143</volume><issue>12</issue><spage>3775</spage><epage>3783</epage><pages>3775-3783</pages><issn>1350-0872</issn><eissn>1465-2080</eissn><abstract>School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK
Author for correspondence: Stephen Spiro. Tel: + 44 1603 592777. Fax: +44 1603 592250. e-mail: s.spiro@uea.ac.uk
ABSTRACT
Paracoccus denitrificans is a heterotrophic organism capable of oxidizing ammonia to nitrite during growth on an organic carbon and energy source. This pathway, termed heterotrophic nitrification, requires the concerted action of an ammonia monooxygenase (AMO) and hydroxylamine oxidase (HAO). The genes required for heterotrophic nitrification have been isolated by introducing a Pa. denitrificans genomic library into Pseudomonas putida and screening for the accumulation of nitrite. In contrast to the situation in chemolithoautotrophic ammonia oxidizers, the genes encoding AMO and HAO are present in single linked copies in the genome of Pa. denitrificans. AMO from Pa. denitrificans expressed in Ps. putida is capable of oxidizing ethene (ethylene) to epoxyethane (ethylene oxide), which is indicative of a relaxed substrate specificity. Further, when expressed in the methylotroph Methylobacterium extorquens AM1, the AMO endows on this organism the ability to grow on ethene and methane. Thus, the Pa. denitrificans AMO is capable of oxidizing methane to methanol, as is the case for the AMO from Nitrosomonas europaea. The heterotrophic nitrification genes are moderately toxic in M. extorquens, more toxic in Ps. putida, and non-toxic in Escherichia coli. Toxicity is due to the activity of the gene products in M. extorquens, and both expression and activity in Ps. putida. This is the first time that the genes encoding an active AMO have been expressed in a heterologous host.
Keywords: heterotrophic nitrification, ammonia monooxygenase, hydroxylamine oxidoreductase, Paracoccus denitrificans, Methylobacterium extorquens
Present address: Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.
Present address: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, UK.</abstract><cop>Reading</cop><pub>Soc General Microbiol</pub><pmid>9421902</pmid><doi>10.1099/00221287-143-12-3775</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; PubMed Central |
| subjects | Ammonia - metabolism Bacteriology Biological and medical sciences Cloning, Molecular - methods Cosmids Fundamental and applied biological sciences. Psychology Genes, Bacterial Genetics Genomic Library Methylobacterium extorquens Microbiology Nitrate Reductase Nitrate Reductases - metabolism Nitrites - metabolism Oxidoreductases - genetics Oxidoreductases - metabolism Paracoccus denitrificans Paracoccus denitrificans - genetics Paracoccus denitrificans - growth & development Paracoccus denitrificans - metabolism Pseudomonas putida - growth & development Pseudomonas putida - metabolism Recombinant Proteins - metabolism Restriction Mapping |
| title | Heterologous expression of heterotrophic nitrification genes |
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