Identification and characterization of a meta-cleavage product hydrolase involved in biphenyl degradation from Arthrobacter sp. YC-RL1
Polychlorinated biphenyls (PCBs) are a group of persistent organic pollutants (POPs) widely existing in the environment. Arthrobacter sp. YC-RL1 is a biphenyl-degrading bacterium that shows metabolic versatility towards aromatic compounds. A 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoate (HOPDA) hydrola...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2019-08, Vol.103 (16), p.6825-6836 |
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| creator | Jia, Yang Wang, Junhuan Ren, Chao Nahurira, Ruth Khokhar, Ibatsam Wang, Jiayi Fan, Shuanghu Yan, Yanchun |
| description | Polychlorinated biphenyls (PCBs) are a group of persistent organic pollutants (POPs) widely existing in the environment.
Arthrobacter
sp. YC-RL1 is a biphenyl-degrading bacterium that shows metabolic versatility towards aromatic compounds. A 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoate (HOPDA) hydrolase (BphD) gene involved in the biodegradation of biphenyl was cloned from strain YC-RL1 and heterologously expressed in
Escherichia coli
BL21 (DE3). The recombinant BphD
YC-RL1
was purified and characterized. BphD
YC-RL1
showed the highest activity at 45 °C and pH 7. It was stable under a wide range of temperature (20–50 °C). The enzyme had a
K
m
value of 0.14 mM,
K
cat
of 11.61 s
−1
, and
V
max
of 0.027 U/mg. Temperature dependence catalysis exhibited a biphasic Arrhenius Plot with a transition at 20 °C. BphD
YC-RL1
was inactivated by SDS, Tween 20, Tween 80, Trition X-100, DTT, CHAPS, NBS, PMSF, and DEPC, but insensitive to EDTA. Site-directed mutagenesis of the active-site residues revealed that the catalytic triad residues (Ser115, His275, and Asp247) of BphD
YC-RL1
were necessary for its activity. The investigation of BphD
YC-RL1
not only provides new potential enzyme resource for the biodegradation of biphenyl but also helps deepen our understanding on the catalytic process and mechanism. |
| doi_str_mv | 10.1007/s00253-019-09956-z |
| format | Article |
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Arthrobacter
sp. YC-RL1 is a biphenyl-degrading bacterium that shows metabolic versatility towards aromatic compounds. A 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoate (HOPDA) hydrolase (BphD) gene involved in the biodegradation of biphenyl was cloned from strain YC-RL1 and heterologously expressed in
Escherichia coli
BL21 (DE3). The recombinant BphD
YC-RL1
was purified and characterized. BphD
YC-RL1
showed the highest activity at 45 °C and pH 7. It was stable under a wide range of temperature (20–50 °C). The enzyme had a
K
m
value of 0.14 mM,
K
cat
of 11.61 s
−1
, and
V
max
of 0.027 U/mg. Temperature dependence catalysis exhibited a biphasic Arrhenius Plot with a transition at 20 °C. BphD
YC-RL1
was inactivated by SDS, Tween 20, Tween 80, Trition X-100, DTT, CHAPS, NBS, PMSF, and DEPC, but insensitive to EDTA. Site-directed mutagenesis of the active-site residues revealed that the catalytic triad residues (Ser115, His275, and Asp247) of BphD
YC-RL1
were necessary for its activity. The investigation of BphD
YC-RL1
not only provides new potential enzyme resource for the biodegradation of biphenyl but also helps deepen our understanding on the catalytic process and mechanism.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-019-09956-z</identifier><identifier>PMID: 31240368</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Aromatic compounds ; Arthrobacter ; Arthrobacter - enzymology ; Arthrobacter - genetics ; Biodegradation ; Biomedical and Life Sciences ; Biotechnology ; Biotransformation ; Biphenyl ; Biphenyl Compounds - metabolism ; Catalysis ; Catalytic Domain ; Cloning, Molecular ; Dithiothreitol ; DNA Mutational Analysis ; E coli ; Environmental Biotechnology ; Enzyme Inhibitors - analysis ; Enzyme Stability ; Enzymes ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Ethylenediaminetetraacetic acid ; Ethylenediaminetetraacetic acids ; Fungicides, Industrial - metabolism ; Gene Expression ; Hydrogen-Ion Concentration ; Hydrolase ; Hydrolases ; Hydrolases - genetics ; Hydrolases - metabolism ; Kinetics ; Life Sciences ; Microbial Genetics and Genomics ; Microbiology ; Mutagenesis, Site-Directed ; PCB ; Persistent organic pollutants ; Pollutants ; Polychlorinated biphenyls ; Residues ; Site-directed mutagenesis ; Surface active agents ; Temperature ; Temperature dependence</subject><ispartof>Applied microbiology and biotechnology, 2019-08, Vol.103 (16), p.6825-6836</ispartof><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2019</rights><rights>COPYRIGHT 2019 Springer</rights><rights>Applied Microbiology and Biotechnology is a copyright of Springer, (2019). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c513t-90820fa9fce3c5a500339d4a6f294519ef3ad21de53d3f5c8a0b03a4873314903</citedby><cites>FETCH-LOGICAL-c513t-90820fa9fce3c5a500339d4a6f294519ef3ad21de53d3f5c8a0b03a4873314903</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-019-09956-z$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-019-09956-z$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31240368$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jia, Yang</creatorcontrib><creatorcontrib>Wang, Junhuan</creatorcontrib><creatorcontrib>Ren, Chao</creatorcontrib><creatorcontrib>Nahurira, Ruth</creatorcontrib><creatorcontrib>Khokhar, Ibatsam</creatorcontrib><creatorcontrib>Wang, Jiayi</creatorcontrib><creatorcontrib>Fan, Shuanghu</creatorcontrib><creatorcontrib>Yan, Yanchun</creatorcontrib><title>Identification and characterization of a meta-cleavage product hydrolase involved in biphenyl degradation from Arthrobacter sp. YC-RL1</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>Polychlorinated biphenyls (PCBs) are a group of persistent organic pollutants (POPs) widely existing in the environment.
Arthrobacter
sp. YC-RL1 is a biphenyl-degrading bacterium that shows metabolic versatility towards aromatic compounds. A 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoate (HOPDA) hydrolase (BphD) gene involved in the biodegradation of biphenyl was cloned from strain YC-RL1 and heterologously expressed in
Escherichia coli
BL21 (DE3). The recombinant BphD
YC-RL1
was purified and characterized. BphD
YC-RL1
showed the highest activity at 45 °C and pH 7. It was stable under a wide range of temperature (20–50 °C). The enzyme had a
K
m
value of 0.14 mM,
K
cat
of 11.61 s
−1
, and
V
max
of 0.027 U/mg. Temperature dependence catalysis exhibited a biphasic Arrhenius Plot with a transition at 20 °C. BphD
YC-RL1
was inactivated by SDS, Tween 20, Tween 80, Trition X-100, DTT, CHAPS, NBS, PMSF, and DEPC, but insensitive to EDTA. Site-directed mutagenesis of the active-site residues revealed that the catalytic triad residues (Ser115, His275, and Asp247) of BphD
YC-RL1
were necessary for its activity. The investigation of BphD
YC-RL1
not only provides new potential enzyme resource for the biodegradation of biphenyl but also helps deepen our understanding on the catalytic process and mechanism.</description><subject>Aromatic compounds</subject><subject>Arthrobacter</subject><subject>Arthrobacter - enzymology</subject><subject>Arthrobacter - genetics</subject><subject>Biodegradation</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Biotransformation</subject><subject>Biphenyl</subject><subject>Biphenyl Compounds - metabolism</subject><subject>Catalysis</subject><subject>Catalytic Domain</subject><subject>Cloning, Molecular</subject><subject>Dithiothreitol</subject><subject>DNA Mutational Analysis</subject><subject>E coli</subject><subject>Environmental Biotechnology</subject><subject>Enzyme Inhibitors - analysis</subject><subject>Enzyme Stability</subject><subject>Enzymes</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Ethylenediaminetetraacetic acid</subject><subject>Ethylenediaminetetraacetic acids</subject><subject>Fungicides, Industrial - metabolism</subject><subject>Gene Expression</subject><subject>Hydrogen-Ion Concentration</subject><subject>Hydrolase</subject><subject>Hydrolases</subject><subject>Hydrolases - genetics</subject><subject>Hydrolases - metabolism</subject><subject>Kinetics</subject><subject>Life Sciences</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Mutagenesis, Site-Directed</subject><subject>PCB</subject><subject>Persistent organic pollutants</subject><subject>Pollutants</subject><subject>Polychlorinated biphenyls</subject><subject>Residues</subject><subject>Site-directed mutagenesis</subject><subject>Surface active agents</subject><subject>Temperature</subject><subject>Temperature dependence</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp9kc1uEzEUhUcIREPhBVggS6xYTLi2xzPjZRSVEikSUoEFK8vxT-JqZhxsT0TyADx33U6hioSQF7auv3Pse09RvMUwxwDNxwhAGC0B8xI4Z3V5elbMcEVJCTWunhczwA0rG8bbi-JVjLcAmLR1_bK4oJhUQOt2VvxeaTMkZ52SyfkByUEjtZNBqmSCO01Fb5FEvUmyVJ2RB7k1aB-8HlVCu6MOvpPRIDccfHcwOh_Qxu13Zjh2SJttkHpyscH3aBHSLvjNgz2K-zn6sSxv1vh18cLKLpo3j_tl8f3T1bfl53L95Xq1XKxLxTBNJYeWgJXcKkMVkwyAUq4rWVvCK4a5sVRqgrVhVFPLVCthA1RWbUMprjjQy-L95Jv__3M0MYlbP4YhPykIqWpW5RnVT9RWdka4wfqUB9K7qMSCcQYNaZp7r_k_qLy06Z3yg7Eu188EH84EmUnmV9rKMUax-npzzpKJVcHHGIwV--B6GY4Cg7hPX0zpi5y-eEhfnLLo3WN346Y3-q_kT9wZoBMQ89WwNeGp_f_Y3gEF07nW</recordid><startdate>20190801</startdate><enddate>20190801</enddate><creator>Jia, Yang</creator><creator>Wang, Junhuan</creator><creator>Ren, Chao</creator><creator>Nahurira, Ruth</creator><creator>Khokhar, Ibatsam</creator><creator>Wang, Jiayi</creator><creator>Fan, Shuanghu</creator><creator>Yan, Yanchun</creator><general>Springer Berlin Heidelberg</general><general>Springer</general><general>Springer Nature B.V</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>ISR</scope><scope>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7WY</scope><scope>7WZ</scope><scope>7X7</scope><scope>7XB</scope><scope>87Z</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8FL</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BEZIV</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FRNLG</scope><scope>FYUFA</scope><scope>F~G</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K60</scope><scope>K6~</scope><scope>K9.</scope><scope>L.-</scope><scope>LK8</scope><scope>M0C</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQBIZ</scope><scope>PQBZA</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope></search><sort><creationdate>20190801</creationdate><title>Identification and characterization of a meta-cleavage product hydrolase involved in biphenyl degradation from Arthrobacter sp. YC-RL1</title><author>Jia, Yang ; Wang, Junhuan ; Ren, Chao ; Nahurira, Ruth ; Khokhar, Ibatsam ; Wang, Jiayi ; Fan, Shuanghu ; Yan, Yanchun</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c513t-90820fa9fce3c5a500339d4a6f294519ef3ad21de53d3f5c8a0b03a4873314903</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Aromatic compounds</topic><topic>Arthrobacter</topic><topic>Arthrobacter - enzymology</topic><topic>Arthrobacter - genetics</topic><topic>Biodegradation</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Biotransformation</topic><topic>Biphenyl</topic><topic>Biphenyl Compounds - metabolism</topic><topic>Catalysis</topic><topic>Catalytic Domain</topic><topic>Cloning, Molecular</topic><topic>Dithiothreitol</topic><topic>DNA Mutational Analysis</topic><topic>E coli</topic><topic>Environmental Biotechnology</topic><topic>Enzyme Inhibitors - analysis</topic><topic>Enzyme Stability</topic><topic>Enzymes</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>Ethylenediaminetetraacetic acid</topic><topic>Ethylenediaminetetraacetic acids</topic><topic>Fungicides, Industrial - metabolism</topic><topic>Gene Expression</topic><topic>Hydrogen-Ion Concentration</topic><topic>Hydrolase</topic><topic>Hydrolases</topic><topic>Hydrolases - genetics</topic><topic>Hydrolases - metabolism</topic><topic>Kinetics</topic><topic>Life Sciences</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>Mutagenesis, Site-Directed</topic><topic>PCB</topic><topic>Persistent organic pollutants</topic><topic>Pollutants</topic><topic>Polychlorinated biphenyls</topic><topic>Residues</topic><topic>Site-directed mutagenesis</topic><topic>Surface active agents</topic><topic>Temperature</topic><topic>Temperature dependence</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jia, Yang</creatorcontrib><creatorcontrib>Wang, Junhuan</creatorcontrib><creatorcontrib>Ren, Chao</creatorcontrib><creatorcontrib>Nahurira, Ruth</creatorcontrib><creatorcontrib>Khokhar, Ibatsam</creatorcontrib><creatorcontrib>Wang, Jiayi</creatorcontrib><creatorcontrib>Fan, Shuanghu</creatorcontrib><creatorcontrib>Yan, Yanchun</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>ABI/INFORM 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biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jia, Yang</au><au>Wang, Junhuan</au><au>Ren, Chao</au><au>Nahurira, Ruth</au><au>Khokhar, Ibatsam</au><au>Wang, Jiayi</au><au>Fan, Shuanghu</au><au>Yan, Yanchun</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification and characterization of a meta-cleavage product hydrolase involved in biphenyl degradation from Arthrobacter sp. YC-RL1</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2019-08-01</date><risdate>2019</risdate><volume>103</volume><issue>16</issue><spage>6825</spage><epage>6836</epage><pages>6825-6836</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>Polychlorinated biphenyls (PCBs) are a group of persistent organic pollutants (POPs) widely existing in the environment.
Arthrobacter
sp. YC-RL1 is a biphenyl-degrading bacterium that shows metabolic versatility towards aromatic compounds. A 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoate (HOPDA) hydrolase (BphD) gene involved in the biodegradation of biphenyl was cloned from strain YC-RL1 and heterologously expressed in
Escherichia coli
BL21 (DE3). The recombinant BphD
YC-RL1
was purified and characterized. BphD
YC-RL1
showed the highest activity at 45 °C and pH 7. It was stable under a wide range of temperature (20–50 °C). The enzyme had a
K
m
value of 0.14 mM,
K
cat
of 11.61 s
−1
, and
V
max
of 0.027 U/mg. Temperature dependence catalysis exhibited a biphasic Arrhenius Plot with a transition at 20 °C. BphD
YC-RL1
was inactivated by SDS, Tween 20, Tween 80, Trition X-100, DTT, CHAPS, NBS, PMSF, and DEPC, but insensitive to EDTA. Site-directed mutagenesis of the active-site residues revealed that the catalytic triad residues (Ser115, His275, and Asp247) of BphD
YC-RL1
were necessary for its activity. The investigation of BphD
YC-RL1
not only provides new potential enzyme resource for the biodegradation of biphenyl but also helps deepen our understanding on the catalytic process and mechanism.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>31240368</pmid><doi>10.1007/s00253-019-09956-z</doi><tpages>12</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0175-7598 |
| ispartof | Applied microbiology and biotechnology, 2019-08, Vol.103 (16), p.6825-6836 |
| issn | 0175-7598 1432-0614 |
| language | eng |
| recordid | cdi_proquest_journals_2246540016 |
| source | MEDLINE; SpringerLink Journals |
| subjects | Aromatic compounds Arthrobacter Arthrobacter - enzymology Arthrobacter - genetics Biodegradation Biomedical and Life Sciences Biotechnology Biotransformation Biphenyl Biphenyl Compounds - metabolism Catalysis Catalytic Domain Cloning, Molecular Dithiothreitol DNA Mutational Analysis E coli Environmental Biotechnology Enzyme Inhibitors - analysis Enzyme Stability Enzymes Escherichia coli - genetics Escherichia coli - metabolism Ethylenediaminetetraacetic acid Ethylenediaminetetraacetic acids Fungicides, Industrial - metabolism Gene Expression Hydrogen-Ion Concentration Hydrolase Hydrolases Hydrolases - genetics Hydrolases - metabolism Kinetics Life Sciences Microbial Genetics and Genomics Microbiology Mutagenesis, Site-Directed PCB Persistent organic pollutants Pollutants Polychlorinated biphenyls Residues Site-directed mutagenesis Surface active agents Temperature Temperature dependence |
| title | Identification and characterization of a meta-cleavage product hydrolase involved in biphenyl degradation from Arthrobacter sp. YC-RL1 |
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