Purification and characterization of the aromatic desulfinase, 2-(2 ′-hydroxyphenyl)benzenesulfinate desulfinase
2-(2 ′-Hydroxyphenyl)benzenesulfinate desulfinase (HPBS desulfinase) catalyzes the cleavage of the carbon–sulfur bond of 2-(2 ′-hydroxyphenyl)benzenesulfinate (HPBS) to form hydroxybiphenyl and sulfite. This is the final step in the desulfurization of dibenzothiophene, the organosulfur compound used...
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| Veröffentlicht in: | Archives of biochemistry and biophysics 2003-07, Vol.415 (1), p.14-23 |
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| container_title | Archives of biochemistry and biophysics |
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| creator | Watkins, L.M Rodriguez, R Schneider, D Broderick, R Cruz, M Chambers, R Ruckman, E Cody, M Mrachko, G.T |
| description | 2-(2
′-Hydroxyphenyl)benzenesulfinate desulfinase (HPBS desulfinase) catalyzes the cleavage of the carbon–sulfur bond of 2-(2
′-hydroxyphenyl)benzenesulfinate (HPBS) to form hydroxybiphenyl and sulfite. This is the final step in the desulfurization of dibenzothiophene, the organosulfur compound used to study biodesulfurization of petroleum middle distillate. HPBS desulfinase was purified 1600-fold from
Rhodococcus IGTS8. The purification was monitored using a spectrofluorimetric assay and SDS–PAGE. The p
I of HPBS desulfinase is 5.6, the temperature optimum is 35
°C, and the pH optimum is 7.0. HPBS desulfinase has a
K
m of 0.90
±
0.15
μM and a
k
cat of 1.3
±
0.07
min
−1. Several analogs were tested for their ability to act as substrates or inhibitors of HPBS desulfinase. No alternative substrates and very few inhibitors were identified. HPBS desulfinase activity decreases in the presence of Cu
2+ and Zn
2+, while no metals significantly enhance enzyme activity. HPBS desulfinase is susceptible to tyrosine, tryptophan, and cysteine specific modification agents. |
| doi_str_mv | 10.1016/S0003-9861(03)00230-3 |
| format | Article |
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′-Hydroxyphenyl)benzenesulfinate desulfinase (HPBS desulfinase) catalyzes the cleavage of the carbon–sulfur bond of 2-(2
′-hydroxyphenyl)benzenesulfinate (HPBS) to form hydroxybiphenyl and sulfite. This is the final step in the desulfurization of dibenzothiophene, the organosulfur compound used to study biodesulfurization of petroleum middle distillate. HPBS desulfinase was purified 1600-fold from
Rhodococcus IGTS8. The purification was monitored using a spectrofluorimetric assay and SDS–PAGE. The p
I of HPBS desulfinase is 5.6, the temperature optimum is 35
°C, and the pH optimum is 7.0. HPBS desulfinase has a
K
m of 0.90
±
0.15
μM and a
k
cat of 1.3
±
0.07
min
−1. Several analogs were tested for their ability to act as substrates or inhibitors of HPBS desulfinase. No alternative substrates and very few inhibitors were identified. HPBS desulfinase activity decreases in the presence of Cu
2+ and Zn
2+, while no metals significantly enhance enzyme activity. HPBS desulfinase is susceptible to tyrosine, tryptophan, and cysteine specific modification agents.</description><identifier>ISSN: 0003-9861</identifier><identifier>EISSN: 1096-0384</identifier><identifier>DOI: 10.1016/S0003-9861(03)00230-3</identifier><identifier>PMID: 12801508</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Cells, Cultured ; Desulfurization ; Enzyme Activation ; Enzyme Inhibitors - chemistry ; Enzyme Stability ; Hydrogen-Ion Concentration ; Inhibition studies ; Kinetics ; Oxidoreductases - antagonists & inhibitors ; Oxidoreductases - chemistry ; Oxidoreductases - isolation & purification ; Oxidoreductases - metabolism ; Quality Control ; Rhodococcus - chemistry ; Rhodococcus - classification ; Rhodococcus - enzymology ; Rhodococcus - isolation & purification ; Rhodococcus erythropolis IGTS8 ; Species Specificity ; Substrate analogs ; Temperature</subject><ispartof>Archives of biochemistry and biophysics, 2003-07, Vol.415 (1), p.14-23</ispartof><rights>2003</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c427t-1b86e6f9409f72456b0ff34e16b99007583464235981b2537490e5ed374e53c3</citedby><cites>FETCH-LOGICAL-c427t-1b86e6f9409f72456b0ff34e16b99007583464235981b2537490e5ed374e53c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0003986103002303$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12801508$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Watkins, L.M</creatorcontrib><creatorcontrib>Rodriguez, R</creatorcontrib><creatorcontrib>Schneider, D</creatorcontrib><creatorcontrib>Broderick, R</creatorcontrib><creatorcontrib>Cruz, M</creatorcontrib><creatorcontrib>Chambers, R</creatorcontrib><creatorcontrib>Ruckman, E</creatorcontrib><creatorcontrib>Cody, M</creatorcontrib><creatorcontrib>Mrachko, G.T</creatorcontrib><title>Purification and characterization of the aromatic desulfinase, 2-(2 ′-hydroxyphenyl)benzenesulfinate desulfinase</title><title>Archives of biochemistry and biophysics</title><addtitle>Arch Biochem Biophys</addtitle><description>2-(2
′-Hydroxyphenyl)benzenesulfinate desulfinase (HPBS desulfinase) catalyzes the cleavage of the carbon–sulfur bond of 2-(2
′-hydroxyphenyl)benzenesulfinate (HPBS) to form hydroxybiphenyl and sulfite. This is the final step in the desulfurization of dibenzothiophene, the organosulfur compound used to study biodesulfurization of petroleum middle distillate. HPBS desulfinase was purified 1600-fold from
Rhodococcus IGTS8. The purification was monitored using a spectrofluorimetric assay and SDS–PAGE. The p
I of HPBS desulfinase is 5.6, the temperature optimum is 35
°C, and the pH optimum is 7.0. HPBS desulfinase has a
K
m of 0.90
±
0.15
μM and a
k
cat of 1.3
±
0.07
min
−1. Several analogs were tested for their ability to act as substrates or inhibitors of HPBS desulfinase. No alternative substrates and very few inhibitors were identified. HPBS desulfinase activity decreases in the presence of Cu
2+ and Zn
2+, while no metals significantly enhance enzyme activity. HPBS desulfinase is susceptible to tyrosine, tryptophan, and cysteine specific modification agents.</description><subject>Cells, Cultured</subject><subject>Desulfurization</subject><subject>Enzyme Activation</subject><subject>Enzyme Inhibitors - chemistry</subject><subject>Enzyme Stability</subject><subject>Hydrogen-Ion Concentration</subject><subject>Inhibition studies</subject><subject>Kinetics</subject><subject>Oxidoreductases - antagonists & inhibitors</subject><subject>Oxidoreductases - chemistry</subject><subject>Oxidoreductases - isolation & purification</subject><subject>Oxidoreductases - metabolism</subject><subject>Quality Control</subject><subject>Rhodococcus - chemistry</subject><subject>Rhodococcus - classification</subject><subject>Rhodococcus - enzymology</subject><subject>Rhodococcus - isolation & purification</subject><subject>Rhodococcus erythropolis IGTS8</subject><subject>Species Specificity</subject><subject>Substrate analogs</subject><subject>Temperature</subject><issn>0003-9861</issn><issn>1096-0384</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkNtKw0AQhhdRbD08gpIrsWB09pBNciVSPEFBQe-XzWaWrqRJ3U3EeuUz-Ug-idHWw51XMwzfP8N8hOxROKZA5ckdAPA4zyQ9BD4CYBxivkaGFHIZA8_EOhn-IAOyFcIDAKVCsk0yoCwDmkA2JP628846o1vX1JGuy8hMtdemRe9elsPGRu0UI-2bWT8wUYmhq6yrdcCjiMWHLHp_fYuni9I3z4v5FOtFNSqwfsH6G2zxb2iHbFhdBdxd1W1yf3F-P76KJzeX1-OzSWwES9uYFplEaXMBuU2ZSGQB1nKBVBZ5DpAmGRdSMJ7kGS1YwlORAyZY9g0m3PBtcrBcO_fNY4ehVTMXDFaVrrHpgko5lymTaQ8mS9D4JgSPVs29m2m_UBTUp2v15Vp9ilR9_XKteJ_bXx3oihmWv6mV3B44XQLYf_nk0KtgHNYGS-fRtKps3D8nPgAk7Y-O</recordid><startdate>20030701</startdate><enddate>20030701</enddate><creator>Watkins, L.M</creator><creator>Rodriguez, R</creator><creator>Schneider, D</creator><creator>Broderick, R</creator><creator>Cruz, M</creator><creator>Chambers, R</creator><creator>Ruckman, E</creator><creator>Cody, M</creator><creator>Mrachko, G.T</creator><general>Elsevier Inc</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>7X8</scope></search><sort><creationdate>20030701</creationdate><title>Purification and characterization of the aromatic desulfinase, 2-(2 ′-hydroxyphenyl)benzenesulfinate desulfinase</title><author>Watkins, L.M ; Rodriguez, R ; Schneider, D ; Broderick, R ; Cruz, M ; Chambers, R ; Ruckman, E ; Cody, M ; Mrachko, G.T</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c427t-1b86e6f9409f72456b0ff34e16b99007583464235981b2537490e5ed374e53c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Cells, Cultured</topic><topic>Desulfurization</topic><topic>Enzyme Activation</topic><topic>Enzyme Inhibitors - chemistry</topic><topic>Enzyme Stability</topic><topic>Hydrogen-Ion Concentration</topic><topic>Inhibition studies</topic><topic>Kinetics</topic><topic>Oxidoreductases - antagonists & inhibitors</topic><topic>Oxidoreductases - chemistry</topic><topic>Oxidoreductases - isolation & purification</topic><topic>Oxidoreductases - metabolism</topic><topic>Quality Control</topic><topic>Rhodococcus - chemistry</topic><topic>Rhodococcus - classification</topic><topic>Rhodococcus - enzymology</topic><topic>Rhodococcus - isolation & purification</topic><topic>Rhodococcus erythropolis IGTS8</topic><topic>Species Specificity</topic><topic>Substrate analogs</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Watkins, L.M</creatorcontrib><creatorcontrib>Rodriguez, R</creatorcontrib><creatorcontrib>Schneider, D</creatorcontrib><creatorcontrib>Broderick, R</creatorcontrib><creatorcontrib>Cruz, M</creatorcontrib><creatorcontrib>Chambers, R</creatorcontrib><creatorcontrib>Ruckman, E</creatorcontrib><creatorcontrib>Cody, M</creatorcontrib><creatorcontrib>Mrachko, G.T</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>Archives of biochemistry and biophysics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Watkins, L.M</au><au>Rodriguez, R</au><au>Schneider, D</au><au>Broderick, R</au><au>Cruz, M</au><au>Chambers, R</au><au>Ruckman, E</au><au>Cody, M</au><au>Mrachko, G.T</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification and characterization of the aromatic desulfinase, 2-(2 ′-hydroxyphenyl)benzenesulfinate desulfinase</atitle><jtitle>Archives of biochemistry and biophysics</jtitle><addtitle>Arch Biochem Biophys</addtitle><date>2003-07-01</date><risdate>2003</risdate><volume>415</volume><issue>1</issue><spage>14</spage><epage>23</epage><pages>14-23</pages><issn>0003-9861</issn><eissn>1096-0384</eissn><abstract>2-(2
′-Hydroxyphenyl)benzenesulfinate desulfinase (HPBS desulfinase) catalyzes the cleavage of the carbon–sulfur bond of 2-(2
′-hydroxyphenyl)benzenesulfinate (HPBS) to form hydroxybiphenyl and sulfite. This is the final step in the desulfurization of dibenzothiophene, the organosulfur compound used to study biodesulfurization of petroleum middle distillate. HPBS desulfinase was purified 1600-fold from
Rhodococcus IGTS8. The purification was monitored using a spectrofluorimetric assay and SDS–PAGE. The p
I of HPBS desulfinase is 5.6, the temperature optimum is 35
°C, and the pH optimum is 7.0. HPBS desulfinase has a
K
m of 0.90
±
0.15
μM and a
k
cat of 1.3
±
0.07
min
−1. Several analogs were tested for their ability to act as substrates or inhibitors of HPBS desulfinase. No alternative substrates and very few inhibitors were identified. HPBS desulfinase activity decreases in the presence of Cu
2+ and Zn
2+, while no metals significantly enhance enzyme activity. HPBS desulfinase is susceptible to tyrosine, tryptophan, and cysteine specific modification agents.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>12801508</pmid><doi>10.1016/S0003-9861(03)00230-3</doi><tpages>10</tpages></addata></record> |
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| subjects | Cells, Cultured Desulfurization Enzyme Activation Enzyme Inhibitors - chemistry Enzyme Stability Hydrogen-Ion Concentration Inhibition studies Kinetics Oxidoreductases - antagonists & inhibitors Oxidoreductases - chemistry Oxidoreductases - isolation & purification Oxidoreductases - metabolism Quality Control Rhodococcus - chemistry Rhodococcus - classification Rhodococcus - enzymology Rhodococcus - isolation & purification Rhodococcus erythropolis IGTS8 Species Specificity Substrate analogs Temperature |
| title | Purification and characterization of the aromatic desulfinase, 2-(2 ′-hydroxyphenyl)benzenesulfinate desulfinase |
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