Production, partial characterization, and potential diagnostic use of salicylate hydroxylase from Pseudomonas putida UUC-1
An unusual strain of Pseudomonas putida UUC-1 capable of growth at high salicylate concentration (10 g l −1) was investigated with the aim of developing an assay and a biosensor system for determining salicylate in body fluids by utilizing the salicylate hydroxylase enzyme. Medium and growth conditi...
Gespeichert in:
| Veröffentlicht in: | Enzyme and microbial technology 1994, Vol.16 (8), p.665-670 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 670 |
|---|---|
| container_issue | 8 |
| container_start_page | 665 |
| container_title | Enzyme and microbial technology |
| container_volume | 16 |
| creator | Banat, I.M. Marchant, A. Nigam, P. Gaston, S.J.S. Kelly, B.A. Marchant, R. |
| description | An unusual strain of
Pseudomonas putida UUC-1 capable of growth at high salicylate concentration (10 g l
−1) was investigated with the aim of developing an assay and a biosensor system for determining salicylate in body fluids by utilizing the salicylate hydroxylase enzyme. Medium and growth condition optimization were carried out under chemostat conditions. The highest biomass yield was at 4.0 g l
−1 salicylate, 25°C, pH 6.5, and 0.2 h
−1 dilution rate. Growth occurred at up to 0.45 h
−1 dilution rate, producing 236 U l
−1 enzyme activity and an output of 424 U h
−1. The activity and productivity were higher than any reported in the literature for this enzyme. It had a
K
m value of 2.07 ± 0.32
μM and an M
r of approximately 43,000. In addition, its specific activity in the crude extract (0.8–0.9 U mg
−1 protein) was similar to the commercially available enzyme. No plasmid DNA was detected in this strain, and no salicylate-negative isolates were obtained when curing with mitomycin C. It is therefore proposed that our strain has a chromosomally located inducible salicylate hydroxylase gene that enables it to grow at high salicylate. This strain also offers a means of cheaply producing large quantities of the enzyme through standard fermentation techniques. |
| doi_str_mv | 10.1016/0141-0229(94)90087-6 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_76636452</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>0141022994900876</els_id><sourcerecordid>16693050</sourcerecordid><originalsourceid>FETCH-LOGICAL-c484t-5338b845d96ec44ddb68819f51fe94a42f5eef87d8c0766bbd51eb70e05859a83</originalsourceid><addsrcrecordid>eNqNkc2KFDEUhYMoYzv6BgpZiChYetOV340gjX8w4CzsdUglt5xIVaVNUmLP01s93fRSXSXk-87lkkPIUwZvGDD5FhhnDazX5qXhrwyAVo28R1ZMK9OAAXOfrM7KQ_KolB8AywOHC3KhlBSg1IrcXucUZl9jml7Tncs1uoH6G5edr5jjrTsSNwW6SxWnOx6i-z6lUqOnc0GaelrcEP1-cBXpzT7k9Hu5L6TPaaTXBeeQxjS5QndzjcHR7XbTsMfkQe-Ggk9O5yXZfvzwbfO5ufr66cvm_VXjuea1EW2rO81FMBI95yF0UmtmesF6NNzxdS8Qe62C9qCk7LogGHYKEIQWxun2krw4zt3l9HPGUu0Yi8dhcBOmudgl1Eou1v8UmTTAW9D_IUrTgoBF5EfR51RKxt7uchxd3lsG9lCiPTRkDw1Zw-1diVYusWen-XM3YjiHTq0t_PmJu-Ld0Gc3-VjOGmeKGdUu2rujhsvv_oqYbfERJ48hZvTVhhT_vscf4Qy5jg</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16693050</pqid></control><display><type>article</type><title>Production, partial characterization, and potential diagnostic use of salicylate hydroxylase from Pseudomonas putida UUC-1</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><creator>Banat, I.M. ; Marchant, A. ; Nigam, P. ; Gaston, S.J.S. ; Kelly, B.A. ; Marchant, R.</creator><creatorcontrib>Banat, I.M. ; Marchant, A. ; Nigam, P. ; Gaston, S.J.S. ; Kelly, B.A. ; Marchant, R.</creatorcontrib><description>An unusual strain of
Pseudomonas putida UUC-1 capable of growth at high salicylate concentration (10 g l
−1) was investigated with the aim of developing an assay and a biosensor system for determining salicylate in body fluids by utilizing the salicylate hydroxylase enzyme. Medium and growth condition optimization were carried out under chemostat conditions. The highest biomass yield was at 4.0 g l
−1 salicylate, 25°C, pH 6.5, and 0.2 h
−1 dilution rate. Growth occurred at up to 0.45 h
−1 dilution rate, producing 236 U l
−1 enzyme activity and an output of 424 U h
−1. The activity and productivity were higher than any reported in the literature for this enzyme. It had a
K
m value of 2.07 ± 0.32
μM and an M
r of approximately 43,000. In addition, its specific activity in the crude extract (0.8–0.9 U mg
−1 protein) was similar to the commercially available enzyme. No plasmid DNA was detected in this strain, and no salicylate-negative isolates were obtained when curing with mitomycin C. It is therefore proposed that our strain has a chromosomally located inducible salicylate hydroxylase gene that enables it to grow at high salicylate. This strain also offers a means of cheaply producing large quantities of the enzyme through standard fermentation techniques.</description><identifier>ISSN: 0141-0229</identifier><identifier>EISSN: 1879-0909</identifier><identifier>DOI: 10.1016/0141-0229(94)90087-6</identifier><identifier>PMID: 7765077</identifier><identifier>CODEN: EMTED2</identifier><language>eng</language><publisher>Amsterdam: Elsevier Inc</publisher><subject>Biological and medical sciences ; Biosensing Techniques ; Biosensors ; Biotechnology ; Culture Media ; Enzyme engineering ; Fermentation ; Fundamental and applied biological sciences. Psychology ; Improved methods for extraction and purification of enzymes ; Methods. Procedures. Technologies ; Mixed Function Oxygenases - biosynthesis ; Mixed Function Oxygenases - isolation & purification ; Plasmids - isolation & purification ; Pseudomonas putida ; Pseudomonas putida - enzymology ; Pseudomonas putida - genetics ; Pseudomonas putida - growth & development ; salicylate ; salicylate hydroxylase ; Salicylates - metabolism ; Salicylic Acid</subject><ispartof>Enzyme and microbial technology, 1994, Vol.16 (8), p.665-670</ispartof><rights>1994</rights><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c484t-5338b845d96ec44ddb68819f51fe94a42f5eef87d8c0766bbd51eb70e05859a83</citedby><cites>FETCH-LOGICAL-c484t-5338b845d96ec44ddb68819f51fe94a42f5eef87d8c0766bbd51eb70e05859a83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/0141022994900876$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,4010,27902,27903,27904,65309</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4171973$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7765077$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Banat, I.M.</creatorcontrib><creatorcontrib>Marchant, A.</creatorcontrib><creatorcontrib>Nigam, P.</creatorcontrib><creatorcontrib>Gaston, S.J.S.</creatorcontrib><creatorcontrib>Kelly, B.A.</creatorcontrib><creatorcontrib>Marchant, R.</creatorcontrib><title>Production, partial characterization, and potential diagnostic use of salicylate hydroxylase from Pseudomonas putida UUC-1</title><title>Enzyme and microbial technology</title><addtitle>Enzyme Microb Technol</addtitle><description>An unusual strain of
Pseudomonas putida UUC-1 capable of growth at high salicylate concentration (10 g l
−1) was investigated with the aim of developing an assay and a biosensor system for determining salicylate in body fluids by utilizing the salicylate hydroxylase enzyme. Medium and growth condition optimization were carried out under chemostat conditions. The highest biomass yield was at 4.0 g l
−1 salicylate, 25°C, pH 6.5, and 0.2 h
−1 dilution rate. Growth occurred at up to 0.45 h
−1 dilution rate, producing 236 U l
−1 enzyme activity and an output of 424 U h
−1. The activity and productivity were higher than any reported in the literature for this enzyme. It had a
K
m value of 2.07 ± 0.32
μM and an M
r of approximately 43,000. In addition, its specific activity in the crude extract (0.8–0.9 U mg
−1 protein) was similar to the commercially available enzyme. No plasmid DNA was detected in this strain, and no salicylate-negative isolates were obtained when curing with mitomycin C. It is therefore proposed that our strain has a chromosomally located inducible salicylate hydroxylase gene that enables it to grow at high salicylate. This strain also offers a means of cheaply producing large quantities of the enzyme through standard fermentation techniques.</description><subject>Biological and medical sciences</subject><subject>Biosensing Techniques</subject><subject>Biosensors</subject><subject>Biotechnology</subject><subject>Culture Media</subject><subject>Enzyme engineering</subject><subject>Fermentation</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Improved methods for extraction and purification of enzymes</subject><subject>Methods. Procedures. Technologies</subject><subject>Mixed Function Oxygenases - biosynthesis</subject><subject>Mixed Function Oxygenases - isolation & purification</subject><subject>Plasmids - isolation & purification</subject><subject>Pseudomonas putida</subject><subject>Pseudomonas putida - enzymology</subject><subject>Pseudomonas putida - genetics</subject><subject>Pseudomonas putida - growth & development</subject><subject>salicylate</subject><subject>salicylate hydroxylase</subject><subject>Salicylates - metabolism</subject><subject>Salicylic Acid</subject><issn>0141-0229</issn><issn>1879-0909</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc2KFDEUhYMoYzv6BgpZiChYetOV340gjX8w4CzsdUglt5xIVaVNUmLP01s93fRSXSXk-87lkkPIUwZvGDD5FhhnDazX5qXhrwyAVo28R1ZMK9OAAXOfrM7KQ_KolB8AywOHC3KhlBSg1IrcXucUZl9jml7Tncs1uoH6G5edr5jjrTsSNwW6SxWnOx6i-z6lUqOnc0GaelrcEP1-cBXpzT7k9Hu5L6TPaaTXBeeQxjS5QndzjcHR7XbTsMfkQe-Ggk9O5yXZfvzwbfO5ufr66cvm_VXjuea1EW2rO81FMBI95yF0UmtmesF6NNzxdS8Qe62C9qCk7LogGHYKEIQWxun2krw4zt3l9HPGUu0Yi8dhcBOmudgl1Eou1v8UmTTAW9D_IUrTgoBF5EfR51RKxt7uchxd3lsG9lCiPTRkDw1Zw-1diVYusWen-XM3YjiHTq0t_PmJu-Ld0Gc3-VjOGmeKGdUu2rujhsvv_oqYbfERJ48hZvTVhhT_vscf4Qy5jg</recordid><startdate>1994</startdate><enddate>1994</enddate><creator>Banat, I.M.</creator><creator>Marchant, A.</creator><creator>Nigam, P.</creator><creator>Gaston, S.J.S.</creator><creator>Kelly, B.A.</creator><creator>Marchant, R.</creator><general>Elsevier Inc</general><general>Elsevier Science</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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>1994</creationdate><title>Production, partial characterization, and potential diagnostic use of salicylate hydroxylase from Pseudomonas putida UUC-1</title><author>Banat, I.M. ; Marchant, A. ; Nigam, P. ; Gaston, S.J.S. ; Kelly, B.A. ; Marchant, R.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c484t-5338b845d96ec44ddb68819f51fe94a42f5eef87d8c0766bbd51eb70e05859a83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Biological and medical sciences</topic><topic>Biosensing Techniques</topic><topic>Biosensors</topic><topic>Biotechnology</topic><topic>Culture Media</topic><topic>Enzyme engineering</topic><topic>Fermentation</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Improved methods for extraction and purification of enzymes</topic><topic>Methods. Procedures. Technologies</topic><topic>Mixed Function Oxygenases - biosynthesis</topic><topic>Mixed Function Oxygenases - isolation & purification</topic><topic>Plasmids - isolation & purification</topic><topic>Pseudomonas putida</topic><topic>Pseudomonas putida - enzymology</topic><topic>Pseudomonas putida - genetics</topic><topic>Pseudomonas putida - growth & development</topic><topic>salicylate</topic><topic>salicylate hydroxylase</topic><topic>Salicylates - metabolism</topic><topic>Salicylic Acid</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Banat, I.M.</creatorcontrib><creatorcontrib>Marchant, A.</creatorcontrib><creatorcontrib>Nigam, P.</creatorcontrib><creatorcontrib>Gaston, S.J.S.</creatorcontrib><creatorcontrib>Kelly, B.A.</creatorcontrib><creatorcontrib>Marchant, R.</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>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Enzyme and microbial technology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Banat, I.M.</au><au>Marchant, A.</au><au>Nigam, P.</au><au>Gaston, S.J.S.</au><au>Kelly, B.A.</au><au>Marchant, R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Production, partial characterization, and potential diagnostic use of salicylate hydroxylase from Pseudomonas putida UUC-1</atitle><jtitle>Enzyme and microbial technology</jtitle><addtitle>Enzyme Microb Technol</addtitle><date>1994</date><risdate>1994</risdate><volume>16</volume><issue>8</issue><spage>665</spage><epage>670</epage><pages>665-670</pages><issn>0141-0229</issn><eissn>1879-0909</eissn><coden>EMTED2</coden><abstract>An unusual strain of
Pseudomonas putida UUC-1 capable of growth at high salicylate concentration (10 g l
−1) was investigated with the aim of developing an assay and a biosensor system for determining salicylate in body fluids by utilizing the salicylate hydroxylase enzyme. Medium and growth condition optimization were carried out under chemostat conditions. The highest biomass yield was at 4.0 g l
−1 salicylate, 25°C, pH 6.5, and 0.2 h
−1 dilution rate. Growth occurred at up to 0.45 h
−1 dilution rate, producing 236 U l
−1 enzyme activity and an output of 424 U h
−1. The activity and productivity were higher than any reported in the literature for this enzyme. It had a
K
m value of 2.07 ± 0.32
μM and an M
r of approximately 43,000. In addition, its specific activity in the crude extract (0.8–0.9 U mg
−1 protein) was similar to the commercially available enzyme. No plasmid DNA was detected in this strain, and no salicylate-negative isolates were obtained when curing with mitomycin C. It is therefore proposed that our strain has a chromosomally located inducible salicylate hydroxylase gene that enables it to grow at high salicylate. This strain also offers a means of cheaply producing large quantities of the enzyme through standard fermentation techniques.</abstract><cop>Amsterdam</cop><pub>Elsevier Inc</pub><pmid>7765077</pmid><doi>10.1016/0141-0229(94)90087-6</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0141-0229 |
| ispartof | Enzyme and microbial technology, 1994, Vol.16 (8), p.665-670 |
| issn | 0141-0229 1879-0909 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_76636452 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Biological and medical sciences Biosensing Techniques Biosensors Biotechnology Culture Media Enzyme engineering Fermentation Fundamental and applied biological sciences. Psychology Improved methods for extraction and purification of enzymes Methods. Procedures. Technologies Mixed Function Oxygenases - biosynthesis Mixed Function Oxygenases - isolation & purification Plasmids - isolation & purification Pseudomonas putida Pseudomonas putida - enzymology Pseudomonas putida - genetics Pseudomonas putida - growth & development salicylate salicylate hydroxylase Salicylates - metabolism Salicylic Acid |
| title | Production, partial characterization, and potential diagnostic use of salicylate hydroxylase from Pseudomonas putida UUC-1 |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-28T01%3A27%3A38IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Production,%20partial%20characterization,%20and%20potential%20diagnostic%20use%20of%20salicylate%20hydroxylase%20from%20Pseudomonas%20putida%20UUC-1&rft.jtitle=Enzyme%20and%20microbial%20technology&rft.au=Banat,%20I.M.&rft.date=1994&rft.volume=16&rft.issue=8&rft.spage=665&rft.epage=670&rft.pages=665-670&rft.issn=0141-0229&rft.eissn=1879-0909&rft.coden=EMTED2&rft_id=info:doi/10.1016/0141-0229(94)90087-6&rft_dat=%3Cproquest_cross%3E16693050%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=16693050&rft_id=info:pmid/7765077&rft_els_id=0141022994900876&rfr_iscdi=true |