6-Hydroxypseudooxynicotine Dehydrogenase Delivers Electrons to Electron Transfer Flavoprotein during Nicotine Degradation by Agrobacterium tumefaciens S33
S33 degrades nicotine via a novel hybrid of the pyridine and the pyrrolidine pathways. The hybrid pathway consists of at least six steps involved in oxidoreductive reactions before the -heterocycle can be broken down. Collectively, the six steps allow electron transfer from nicotine and its intermed...
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| Veröffentlicht in: | Applied and environmental microbiology 2019-06, Vol.85 (11), p.1 |
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| creator | Wang, Rongshui Yi, Jihong Shang, Jinmeng Yu, Wenjun Li, Zhifeng Huang, Haiyan Xie, Huijun Wang, Shuning |
| description | S33 degrades nicotine via a novel hybrid of the pyridine and the pyrrolidine pathways. The hybrid pathway consists of at least six steps involved in oxidoreductive reactions before the
-heterocycle can be broken down. Collectively, the six steps allow electron transfer from nicotine and its intermediates to the final acceptor O
via the electron transport chain (ETC). 6-Hydroxypseudooxynicotine oxidase, renamed 6-hydroxypseudooxynicotine dehydrogenase in this study, has been characterized as catalyzing the fourth step using the artificial electron acceptor 2,6-dichlorophenolindophenol. Here, we used biochemical, genetic, and liquid chromatography-mass spectrometry (LC-MS) analyses to determine that 6-hydroxypseudooxynicotine dehydrogenase utilizes the electron transfer flavoprotein (EtfAB) as the physiological electron acceptor to catalyze the dehydrogenation of pseudooxynicotine, an analogue of the true substrate 6-hydroxypseudooxynicotine,
, into 3-succinoyl-semialdehyde-pyridine. NAD(P)
, O
, and ferredoxin could not function as electron acceptors. The oxygen atom in the aldehyde group of the product 3-succinoyl-semialdehyde-pyridine was verified to be derived from H
O. Disruption of the
genes in the nicotine-degrading gene cluster decreased the growth rate of
S33 on nicotine but not on 6-hydroxy-3-succinoylpyridine, an intermediate downstream of the hybrid pathway, indicating the requirement of EtfAB for efficient nicotine degradation. The electrons were found to be further transferred from the reduced EtfAB to coenzyme Q by the catalysis of electron transfer flavoprotein:ubiquinone oxidoreductase. These results aid in an in-depth understanding of the electron transfer process and energy metabolism involved in the nicotine oxidation and provide novel insights into nicotine catabolism in bacteria.
Nicotine has been studied as a model for toxic
-heterocyclic aromatic compounds. Microorganisms can catabolize nicotine via various pathways and conserve energy from its oxidation. Although several oxidoreductases have been characterized to participate in nicotine degradation, the electron transfer involved in these processes is poorly understood. In this study, we found that 6-hydroxypseudooxynicotine dehydrogenase, a key enzyme in the hybrid pyridine and pyrrolidine pathway for nicotine degradation in
S33, utilizes EtfAB as a physiological electron acceptor. Catalyzed by the membrane-associated electron transfer flavoprotein:ubiquinone oxidoreductase, the ele |
| doi_str_mv | 10.1128/AEM.00454-19 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6532033</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2231999113</sourcerecordid><originalsourceid>FETCH-LOGICAL-c412t-90d2af5878dcac439d41ffb0eba4d3fb85811246569343025ce92c3c9528b8e3</originalsourceid><addsrcrecordid>eNpVkU1vEzEQhi0EomnhxhlZ4soWf-3WviBFJaVIBQ7kbnnt2a2rjR1sb9T8FX4tTlsCnOzRPH5m5BehN5ScU8rkh-Xq6zkhohUNVc_QghIlm5bz7jlaEKJUw5ggJ-g05ztSMdLJl-iEE8W6CyYX6FfXXO9divf7bYbZxXoJ3sbiA-BPcHtojRBMPlST30HKeDWBLSmGjEs8FnidTMgDJHw1mV3cpljAB-zm5MOIv_1Vjsk4U3x90e_xckyxN7ZA8vMGl3kDg7EeqvoH56_Qi8FMGV4_nWdofbVaX143N98_f7lc3jRWUFYaRRwzQysvpLPGCq6coMPQE-iNcHzoZSvrP4mu7RQXnLDWgmKWW9Uy2UvgZ-jjo3Y79xtwFkJJZtLb5Dcm7XU0Xv_fCf5Wj3Gnu5YzwnkVvHsSpPhzhlz0XZxTqCtrxjhVSlF6oN4_UjbFnBMMxwmU6EOQugapH4LUVFX87b9bHeE_yfHfoZCdpw</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2231999113</pqid></control><display><type>article</type><title>6-Hydroxypseudooxynicotine Dehydrogenase Delivers Electrons to Electron Transfer Flavoprotein during Nicotine Degradation by Agrobacterium tumefaciens S33</title><source>American Society for Microbiology</source><source>PubMed Central Free</source><source>MEDLINE</source><source>Alma/SFX Local Collection</source><creator>Wang, Rongshui ; Yi, Jihong ; Shang, Jinmeng ; Yu, Wenjun ; Li, Zhifeng ; Huang, Haiyan ; Xie, Huijun ; Wang, Shuning</creator><contributor>Parales, Rebecca E.</contributor><creatorcontrib>Wang, Rongshui ; Yi, Jihong ; Shang, Jinmeng ; Yu, Wenjun ; Li, Zhifeng ; Huang, Haiyan ; Xie, Huijun ; Wang, Shuning ; Parales, Rebecca E.</creatorcontrib><description>S33 degrades nicotine via a novel hybrid of the pyridine and the pyrrolidine pathways. The hybrid pathway consists of at least six steps involved in oxidoreductive reactions before the
-heterocycle can be broken down. Collectively, the six steps allow electron transfer from nicotine and its intermediates to the final acceptor O
via the electron transport chain (ETC). 6-Hydroxypseudooxynicotine oxidase, renamed 6-hydroxypseudooxynicotine dehydrogenase in this study, has been characterized as catalyzing the fourth step using the artificial electron acceptor 2,6-dichlorophenolindophenol. Here, we used biochemical, genetic, and liquid chromatography-mass spectrometry (LC-MS) analyses to determine that 6-hydroxypseudooxynicotine dehydrogenase utilizes the electron transfer flavoprotein (EtfAB) as the physiological electron acceptor to catalyze the dehydrogenation of pseudooxynicotine, an analogue of the true substrate 6-hydroxypseudooxynicotine,
, into 3-succinoyl-semialdehyde-pyridine. NAD(P)
, O
, and ferredoxin could not function as electron acceptors. The oxygen atom in the aldehyde group of the product 3-succinoyl-semialdehyde-pyridine was verified to be derived from H
O. Disruption of the
genes in the nicotine-degrading gene cluster decreased the growth rate of
S33 on nicotine but not on 6-hydroxy-3-succinoylpyridine, an intermediate downstream of the hybrid pathway, indicating the requirement of EtfAB for efficient nicotine degradation. The electrons were found to be further transferred from the reduced EtfAB to coenzyme Q by the catalysis of electron transfer flavoprotein:ubiquinone oxidoreductase. These results aid in an in-depth understanding of the electron transfer process and energy metabolism involved in the nicotine oxidation and provide novel insights into nicotine catabolism in bacteria.
Nicotine has been studied as a model for toxic
-heterocyclic aromatic compounds. Microorganisms can catabolize nicotine via various pathways and conserve energy from its oxidation. Although several oxidoreductases have been characterized to participate in nicotine degradation, the electron transfer involved in these processes is poorly understood. In this study, we found that 6-hydroxypseudooxynicotine dehydrogenase, a key enzyme in the hybrid pyridine and pyrrolidine pathway for nicotine degradation in
S33, utilizes EtfAB as a physiological electron acceptor. Catalyzed by the membrane-associated electron transfer flavoprotein:ubiquinone oxidoreductase, the electrons are transferred from the reduced EtfAB to coenzyme Q, which then could enter into the classic ETC. Thus, the route for electron transport from the substrate to O
could be constructed, by which ATP can be further sythesized via chemiosmosis to support the baterial growth. These findings provide new knowledge regarding the catabolism of
-heterocyclic aromatic compounds in microorganisms.</description><identifier>ISSN: 0099-2240</identifier><identifier>EISSN: 1098-5336</identifier><identifier>DOI: 10.1128/AEM.00454-19</identifier><identifier>PMID: 30926728</identifier><language>eng</language><publisher>United States: American Society for Microbiology</publisher><subject>Agrobacterium tumefaciens ; Agrobacterium tumefaciens - genetics ; Agrobacterium tumefaciens - metabolism ; Bacterial Proteins - genetics ; Biodegradation ; Butanones - metabolism ; Catabolism ; Catalysis ; Coenzyme Q ; Degradation ; Dehydrogenase ; Dehydrogenases ; Dehydrogenation ; Dichlorophenolindophenol ; Disruption ; Electron transfer ; Electron transport ; Electron Transport - physiology ; Electron transport chain ; Electron Transport Chain Complex Proteins - metabolism ; Electron-Transferring Flavoproteins - metabolism ; Electrons ; Energy metabolism ; Enzymology and Protein Engineering ; Ferredoxin ; Gene Expression Regulation, Bacterial ; Genetic analysis ; Growth rate ; Intermediates ; Liquid chromatography ; Mass spectrometry ; Mass spectroscopy ; Metabolic Networks and Pathways ; Metabolism ; NAD ; Nicotine ; Nicotine - analogs & derivatives ; Nicotine - metabolism ; Oxidation ; Oxidation-Reduction ; Oxidoreductases - genetics ; Oxidoreductases - metabolism ; Oxygen - metabolism ; Pyridines ; Pyridines - metabolism ; Pyrrolidine ; Recombinant Proteins ; Substrates ; Succinates ; Transcriptome ; Ubiquinone ; Ubiquinone oxidoreductase</subject><ispartof>Applied and environmental microbiology, 2019-06, Vol.85 (11), p.1</ispartof><rights>Copyright © 2019 American Society for Microbiology.</rights><rights>Copyright American Society for Microbiology Jun 2019</rights><rights>Copyright © 2019 American Society for Microbiology. 2019 American Society for Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c412t-90d2af5878dcac439d41ffb0eba4d3fb85811246569343025ce92c3c9528b8e3</citedby><cites>FETCH-LOGICAL-c412t-90d2af5878dcac439d41ffb0eba4d3fb85811246569343025ce92c3c9528b8e3</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/PMC6532033/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6532033/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,3188,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30926728$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Parales, Rebecca E.</contributor><creatorcontrib>Wang, Rongshui</creatorcontrib><creatorcontrib>Yi, Jihong</creatorcontrib><creatorcontrib>Shang, Jinmeng</creatorcontrib><creatorcontrib>Yu, Wenjun</creatorcontrib><creatorcontrib>Li, Zhifeng</creatorcontrib><creatorcontrib>Huang, Haiyan</creatorcontrib><creatorcontrib>Xie, Huijun</creatorcontrib><creatorcontrib>Wang, Shuning</creatorcontrib><title>6-Hydroxypseudooxynicotine Dehydrogenase Delivers Electrons to Electron Transfer Flavoprotein during Nicotine Degradation by Agrobacterium tumefaciens S33</title><title>Applied and environmental microbiology</title><addtitle>Appl Environ Microbiol</addtitle><description>S33 degrades nicotine via a novel hybrid of the pyridine and the pyrrolidine pathways. The hybrid pathway consists of at least six steps involved in oxidoreductive reactions before the
-heterocycle can be broken down. Collectively, the six steps allow electron transfer from nicotine and its intermediates to the final acceptor O
via the electron transport chain (ETC). 6-Hydroxypseudooxynicotine oxidase, renamed 6-hydroxypseudooxynicotine dehydrogenase in this study, has been characterized as catalyzing the fourth step using the artificial electron acceptor 2,6-dichlorophenolindophenol. Here, we used biochemical, genetic, and liquid chromatography-mass spectrometry (LC-MS) analyses to determine that 6-hydroxypseudooxynicotine dehydrogenase utilizes the electron transfer flavoprotein (EtfAB) as the physiological electron acceptor to catalyze the dehydrogenation of pseudooxynicotine, an analogue of the true substrate 6-hydroxypseudooxynicotine,
, into 3-succinoyl-semialdehyde-pyridine. NAD(P)
, O
, and ferredoxin could not function as electron acceptors. The oxygen atom in the aldehyde group of the product 3-succinoyl-semialdehyde-pyridine was verified to be derived from H
O. Disruption of the
genes in the nicotine-degrading gene cluster decreased the growth rate of
S33 on nicotine but not on 6-hydroxy-3-succinoylpyridine, an intermediate downstream of the hybrid pathway, indicating the requirement of EtfAB for efficient nicotine degradation. The electrons were found to be further transferred from the reduced EtfAB to coenzyme Q by the catalysis of electron transfer flavoprotein:ubiquinone oxidoreductase. These results aid in an in-depth understanding of the electron transfer process and energy metabolism involved in the nicotine oxidation and provide novel insights into nicotine catabolism in bacteria.
Nicotine has been studied as a model for toxic
-heterocyclic aromatic compounds. Microorganisms can catabolize nicotine via various pathways and conserve energy from its oxidation. Although several oxidoreductases have been characterized to participate in nicotine degradation, the electron transfer involved in these processes is poorly understood. In this study, we found that 6-hydroxypseudooxynicotine dehydrogenase, a key enzyme in the hybrid pyridine and pyrrolidine pathway for nicotine degradation in
S33, utilizes EtfAB as a physiological electron acceptor. Catalyzed by the membrane-associated electron transfer flavoprotein:ubiquinone oxidoreductase, the electrons are transferred from the reduced EtfAB to coenzyme Q, which then could enter into the classic ETC. Thus, the route for electron transport from the substrate to O
could be constructed, by which ATP can be further sythesized via chemiosmosis to support the baterial growth. These findings provide new knowledge regarding the catabolism of
-heterocyclic aromatic compounds in microorganisms.</description><subject>Agrobacterium tumefaciens</subject><subject>Agrobacterium tumefaciens - genetics</subject><subject>Agrobacterium tumefaciens - metabolism</subject><subject>Bacterial Proteins - genetics</subject><subject>Biodegradation</subject><subject>Butanones - metabolism</subject><subject>Catabolism</subject><subject>Catalysis</subject><subject>Coenzyme Q</subject><subject>Degradation</subject><subject>Dehydrogenase</subject><subject>Dehydrogenases</subject><subject>Dehydrogenation</subject><subject>Dichlorophenolindophenol</subject><subject>Disruption</subject><subject>Electron transfer</subject><subject>Electron transport</subject><subject>Electron Transport - physiology</subject><subject>Electron transport chain</subject><subject>Electron Transport Chain Complex Proteins - metabolism</subject><subject>Electron-Transferring Flavoproteins - metabolism</subject><subject>Electrons</subject><subject>Energy metabolism</subject><subject>Enzymology and Protein Engineering</subject><subject>Ferredoxin</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Genetic analysis</subject><subject>Growth rate</subject><subject>Intermediates</subject><subject>Liquid chromatography</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>Metabolic Networks and Pathways</subject><subject>Metabolism</subject><subject>NAD</subject><subject>Nicotine</subject><subject>Nicotine - analogs & derivatives</subject><subject>Nicotine - metabolism</subject><subject>Oxidation</subject><subject>Oxidation-Reduction</subject><subject>Oxidoreductases - genetics</subject><subject>Oxidoreductases - metabolism</subject><subject>Oxygen - metabolism</subject><subject>Pyridines</subject><subject>Pyridines - metabolism</subject><subject>Pyrrolidine</subject><subject>Recombinant Proteins</subject><subject>Substrates</subject><subject>Succinates</subject><subject>Transcriptome</subject><subject>Ubiquinone</subject><subject>Ubiquinone oxidoreductase</subject><issn>0099-2240</issn><issn>1098-5336</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkU1vEzEQhi0EomnhxhlZ4soWf-3WviBFJaVIBQ7kbnnt2a2rjR1sb9T8FX4tTlsCnOzRPH5m5BehN5ScU8rkh-Xq6zkhohUNVc_QghIlm5bz7jlaEKJUw5ggJ-g05ztSMdLJl-iEE8W6CyYX6FfXXO9divf7bYbZxXoJ3sbiA-BPcHtojRBMPlST30HKeDWBLSmGjEs8FnidTMgDJHw1mV3cpljAB-zm5MOIv_1Vjsk4U3x90e_xckyxN7ZA8vMGl3kDg7EeqvoH56_Qi8FMGV4_nWdofbVaX143N98_f7lc3jRWUFYaRRwzQysvpLPGCq6coMPQE-iNcHzoZSvrP4mu7RQXnLDWgmKWW9Uy2UvgZ-jjo3Y79xtwFkJJZtLb5Dcm7XU0Xv_fCf5Wj3Gnu5YzwnkVvHsSpPhzhlz0XZxTqCtrxjhVSlF6oN4_UjbFnBMMxwmU6EOQugapH4LUVFX87b9bHeE_yfHfoZCdpw</recordid><startdate>20190601</startdate><enddate>20190601</enddate><creator>Wang, Rongshui</creator><creator>Yi, Jihong</creator><creator>Shang, Jinmeng</creator><creator>Yu, Wenjun</creator><creator>Li, Zhifeng</creator><creator>Huang, Haiyan</creator><creator>Xie, Huijun</creator><creator>Wang, Shuning</creator><general>American Society for 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>7QO</scope><scope>7SN</scope><scope>7SS</scope><scope>7ST</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>SOI</scope><scope>5PM</scope></search><sort><creationdate>20190601</creationdate><title>6-Hydroxypseudooxynicotine Dehydrogenase Delivers Electrons to Electron Transfer Flavoprotein during Nicotine Degradation by Agrobacterium tumefaciens S33</title><author>Wang, Rongshui ; Yi, Jihong ; Shang, Jinmeng ; Yu, Wenjun ; Li, Zhifeng ; Huang, Haiyan ; Xie, Huijun ; Wang, Shuning</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c412t-90d2af5878dcac439d41ffb0eba4d3fb85811246569343025ce92c3c9528b8e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Agrobacterium tumefaciens</topic><topic>Agrobacterium tumefaciens - genetics</topic><topic>Agrobacterium tumefaciens - metabolism</topic><topic>Bacterial Proteins - genetics</topic><topic>Biodegradation</topic><topic>Butanones - metabolism</topic><topic>Catabolism</topic><topic>Catalysis</topic><topic>Coenzyme Q</topic><topic>Degradation</topic><topic>Dehydrogenase</topic><topic>Dehydrogenases</topic><topic>Dehydrogenation</topic><topic>Dichlorophenolindophenol</topic><topic>Disruption</topic><topic>Electron transfer</topic><topic>Electron transport</topic><topic>Electron Transport - physiology</topic><topic>Electron transport chain</topic><topic>Electron Transport Chain Complex Proteins - metabolism</topic><topic>Electron-Transferring Flavoproteins - metabolism</topic><topic>Electrons</topic><topic>Energy metabolism</topic><topic>Enzymology and Protein Engineering</topic><topic>Ferredoxin</topic><topic>Gene Expression Regulation, Bacterial</topic><topic>Genetic analysis</topic><topic>Growth rate</topic><topic>Intermediates</topic><topic>Liquid chromatography</topic><topic>Mass spectrometry</topic><topic>Mass spectroscopy</topic><topic>Metabolic Networks and Pathways</topic><topic>Metabolism</topic><topic>NAD</topic><topic>Nicotine</topic><topic>Nicotine - analogs & derivatives</topic><topic>Nicotine - metabolism</topic><topic>Oxidation</topic><topic>Oxidation-Reduction</topic><topic>Oxidoreductases - genetics</topic><topic>Oxidoreductases - metabolism</topic><topic>Oxygen - metabolism</topic><topic>Pyridines</topic><topic>Pyridines - metabolism</topic><topic>Pyrrolidine</topic><topic>Recombinant Proteins</topic><topic>Substrates</topic><topic>Succinates</topic><topic>Transcriptome</topic><topic>Ubiquinone</topic><topic>Ubiquinone oxidoreductase</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Rongshui</creatorcontrib><creatorcontrib>Yi, Jihong</creatorcontrib><creatorcontrib>Shang, Jinmeng</creatorcontrib><creatorcontrib>Yu, Wenjun</creatorcontrib><creatorcontrib>Li, Zhifeng</creatorcontrib><creatorcontrib>Huang, Haiyan</creatorcontrib><creatorcontrib>Xie, Huijun</creatorcontrib><creatorcontrib>Wang, Shuning</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>Biotechnology Research Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Environment Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Environment Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Applied and environmental microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Rongshui</au><au>Yi, Jihong</au><au>Shang, Jinmeng</au><au>Yu, Wenjun</au><au>Li, Zhifeng</au><au>Huang, Haiyan</au><au>Xie, Huijun</au><au>Wang, Shuning</au><au>Parales, Rebecca E.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>6-Hydroxypseudooxynicotine Dehydrogenase Delivers Electrons to Electron Transfer Flavoprotein during Nicotine Degradation by Agrobacterium tumefaciens S33</atitle><jtitle>Applied and environmental microbiology</jtitle><addtitle>Appl Environ Microbiol</addtitle><date>2019-06-01</date><risdate>2019</risdate><volume>85</volume><issue>11</issue><spage>1</spage><pages>1-</pages><issn>0099-2240</issn><eissn>1098-5336</eissn><abstract>S33 degrades nicotine via a novel hybrid of the pyridine and the pyrrolidine pathways. The hybrid pathway consists of at least six steps involved in oxidoreductive reactions before the
-heterocycle can be broken down. Collectively, the six steps allow electron transfer from nicotine and its intermediates to the final acceptor O
via the electron transport chain (ETC). 6-Hydroxypseudooxynicotine oxidase, renamed 6-hydroxypseudooxynicotine dehydrogenase in this study, has been characterized as catalyzing the fourth step using the artificial electron acceptor 2,6-dichlorophenolindophenol. Here, we used biochemical, genetic, and liquid chromatography-mass spectrometry (LC-MS) analyses to determine that 6-hydroxypseudooxynicotine dehydrogenase utilizes the electron transfer flavoprotein (EtfAB) as the physiological electron acceptor to catalyze the dehydrogenation of pseudooxynicotine, an analogue of the true substrate 6-hydroxypseudooxynicotine,
, into 3-succinoyl-semialdehyde-pyridine. NAD(P)
, O
, and ferredoxin could not function as electron acceptors. The oxygen atom in the aldehyde group of the product 3-succinoyl-semialdehyde-pyridine was verified to be derived from H
O. Disruption of the
genes in the nicotine-degrading gene cluster decreased the growth rate of
S33 on nicotine but not on 6-hydroxy-3-succinoylpyridine, an intermediate downstream of the hybrid pathway, indicating the requirement of EtfAB for efficient nicotine degradation. The electrons were found to be further transferred from the reduced EtfAB to coenzyme Q by the catalysis of electron transfer flavoprotein:ubiquinone oxidoreductase. These results aid in an in-depth understanding of the electron transfer process and energy metabolism involved in the nicotine oxidation and provide novel insights into nicotine catabolism in bacteria.
Nicotine has been studied as a model for toxic
-heterocyclic aromatic compounds. Microorganisms can catabolize nicotine via various pathways and conserve energy from its oxidation. Although several oxidoreductases have been characterized to participate in nicotine degradation, the electron transfer involved in these processes is poorly understood. In this study, we found that 6-hydroxypseudooxynicotine dehydrogenase, a key enzyme in the hybrid pyridine and pyrrolidine pathway for nicotine degradation in
S33, utilizes EtfAB as a physiological electron acceptor. Catalyzed by the membrane-associated electron transfer flavoprotein:ubiquinone oxidoreductase, the electrons are transferred from the reduced EtfAB to coenzyme Q, which then could enter into the classic ETC. Thus, the route for electron transport from the substrate to O
could be constructed, by which ATP can be further sythesized via chemiosmosis to support the baterial growth. These findings provide new knowledge regarding the catabolism of
-heterocyclic aromatic compounds in microorganisms.</abstract><cop>United States</cop><pub>American Society for Microbiology</pub><pmid>30926728</pmid><doi>10.1128/AEM.00454-19</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0099-2240 |
| ispartof | Applied and environmental microbiology, 2019-06, Vol.85 (11), p.1 |
| issn | 0099-2240 1098-5336 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6532033 |
| source | American Society for Microbiology; PubMed Central Free; MEDLINE; Alma/SFX Local Collection |
| subjects | Agrobacterium tumefaciens Agrobacterium tumefaciens - genetics Agrobacterium tumefaciens - metabolism Bacterial Proteins - genetics Biodegradation Butanones - metabolism Catabolism Catalysis Coenzyme Q Degradation Dehydrogenase Dehydrogenases Dehydrogenation Dichlorophenolindophenol Disruption Electron transfer Electron transport Electron Transport - physiology Electron transport chain Electron Transport Chain Complex Proteins - metabolism Electron-Transferring Flavoproteins - metabolism Electrons Energy metabolism Enzymology and Protein Engineering Ferredoxin Gene Expression Regulation, Bacterial Genetic analysis Growth rate Intermediates Liquid chromatography Mass spectrometry Mass spectroscopy Metabolic Networks and Pathways Metabolism NAD Nicotine Nicotine - analogs & derivatives Nicotine - metabolism Oxidation Oxidation-Reduction Oxidoreductases - genetics Oxidoreductases - metabolism Oxygen - metabolism Pyridines Pyridines - metabolism Pyrrolidine Recombinant Proteins Substrates Succinates Transcriptome Ubiquinone Ubiquinone oxidoreductase |
| title | 6-Hydroxypseudooxynicotine Dehydrogenase Delivers Electrons to Electron Transfer Flavoprotein during Nicotine Degradation by Agrobacterium tumefaciens S33 |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T13%3A46%3A51IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=6-Hydroxypseudooxynicotine%20Dehydrogenase%20Delivers%20Electrons%20to%20Electron%20Transfer%20Flavoprotein%20during%20Nicotine%20Degradation%20by%20Agrobacterium%20tumefaciens%20S33&rft.jtitle=Applied%20and%20environmental%20microbiology&rft.au=Wang,%20Rongshui&rft.date=2019-06-01&rft.volume=85&rft.issue=11&rft.spage=1&rft.pages=1-&rft.issn=0099-2240&rft.eissn=1098-5336&rft_id=info:doi/10.1128/AEM.00454-19&rft_dat=%3Cproquest_pubme%3E2231999113%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2231999113&rft_id=info:pmid/30926728&rfr_iscdi=true |