Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli
Urate oxidase (EC 1.7.3.3) is an enzyme involved in purine metabolism which is used in the treatment of gout and as diagnostic reagent for detection of uric acid. In order to produce this enzyme in large quantities for biotechnological purposes, the gene coding for the Bacillus subtilis urate oxidas...
Gespeichert in:
| Veröffentlicht in: | BioMed research international 2010-01, Vol.2010 (2010), p.1-6 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 6 |
|---|---|
| container_issue | 2010 |
| container_start_page | 1 |
| container_title | BioMed research international |
| container_volume | 2010 |
| creator | Pfrimer, Pollyana Moraes, Lídia Maria Pepe de Galdino, Alexsandro Sobreira Salles, Loise P. Reis, Viviane Castelo Branco De Marco, Janice Lisboa Prates, Maura Vianna Bloch, Carlos Torres, Fernando Araripe Gonçalves |
| description | Urate oxidase (EC 1.7.3.3) is an enzyme involved in purine metabolism which is used in the treatment of gout and as diagnostic reagent for detection of uric acid. In order to produce this enzyme in large quantities for biotechnological purposes, the gene coding for the Bacillus subtilis urate oxidase was cloned and heterologously expressed in Escherichia coli. Time course induction in E. coli showed an induced protein with an apparent molecular mass of ∼60 kDa. Soluble recombinant enzyme was purified in a single-step procedure using Ni-NTA column. The enzyme was purified 2.1-fold with a yield of 56% compared to the crude extract. MALDI-TOF analysis revealed an ion with a mass of 58675 Da which is in agreement with the expected mass of the recombinant protein. The purified enzyme showed an optimal pH and temperature of 8.0 and 37∘C, respectively, and retained 90% of its activity after 72 hours of incubation at −20∘C and 4∘C. |
| doi_str_mv | 10.1155/2010/674908 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2820260</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2287917791</sourcerecordid><originalsourceid>FETCH-LOGICAL-c425t-f620d9b3251b7f81029c039cbe98d0b15ef54dd3175ca05bf3638c73172aeb3a3</originalsourceid><addsrcrecordid>eNqF0Utv1DAQAOAIgWgpnDiDLCQu0KV-xLF9QYLV8pAqtQd6tiaO3UyVOls7gcKvxyXLAidOtjXfPKypqqeMvmFMyhNOGT1pVG2ovlcdMsboSnHJ7u_vtTioHuV8RSlTujEPq4OS0mij1GE1rYcxYrw8JudzwoAOJhzjMYHYkXNIE8JA1j0kcJNP-ONXlIyBvAeHwzBnkud2wgEzuUgweXJ2ix1kTza32-Rz9h3BSDbZ9SXb9QjEjQM-rh4EGLJ_sjuPqosPmy_rT6vTs4-f1-9OV67mclqFhtPOtKJ8plVBM8qNo8K41hvd0ZZJH2TddYIp6YDKNohGaKfKm4NvBYij6u1Sdzu3175zPk4JBrtNeA3pux0B7b-RiL29HL9arjnlDS0FXuwKpPFm9nmyV-OcYpnZatkwRZnhBb1ekEtjzsmHfQNG7d2G7N2G7LKhop__PdPe_l5JAS93ALKDISSIDvMfJ3XdSCOLe7W4HmMH3_A_XZ8t2BfiA-xxrY2smfgJXH2wnw</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>856170192</pqid></control><display><type>article</type><title>Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli</title><source>MEDLINE</source><source>Wiley-Blackwell Open Access Titles</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><source>PubMed Central Open Access</source><creator>Pfrimer, Pollyana ; Moraes, Lídia Maria Pepe de ; Galdino, Alexsandro Sobreira ; Salles, Loise P. ; Reis, Viviane Castelo Branco ; De Marco, Janice Lisboa ; Prates, Maura Vianna ; Bloch, Carlos ; Torres, Fernando Araripe Gonçalves</creator><contributor>Mowbray, Sherry</contributor><creatorcontrib>Pfrimer, Pollyana ; Moraes, Lídia Maria Pepe de ; Galdino, Alexsandro Sobreira ; Salles, Loise P. ; Reis, Viviane Castelo Branco ; De Marco, Janice Lisboa ; Prates, Maura Vianna ; Bloch, Carlos ; Torres, Fernando Araripe Gonçalves ; Mowbray, Sherry</creatorcontrib><description>Urate oxidase (EC 1.7.3.3) is an enzyme involved in purine metabolism which is used in the treatment of gout and as diagnostic reagent for detection of uric acid. In order to produce this enzyme in large quantities for biotechnological purposes, the gene coding for the Bacillus subtilis urate oxidase was cloned and heterologously expressed in Escherichia coli. Time course induction in E. coli showed an induced protein with an apparent molecular mass of ∼60 kDa. Soluble recombinant enzyme was purified in a single-step procedure using Ni-NTA column. The enzyme was purified 2.1-fold with a yield of 56% compared to the crude extract. MALDI-TOF analysis revealed an ion with a mass of 58675 Da which is in agreement with the expected mass of the recombinant protein. The purified enzyme showed an optimal pH and temperature of 8.0 and 37∘C, respectively, and retained 90% of its activity after 72 hours of incubation at −20∘C and 4∘C.</description><identifier>ISSN: 1110-7243</identifier><identifier>ISSN: 2314-6133</identifier><identifier>EISSN: 1110-7251</identifier><identifier>EISSN: 2314-6141</identifier><identifier>DOI: 10.1155/2010/674908</identifier><identifier>PMID: 20168977</identifier><language>eng</language><publisher>Cairo, Egypt: Hindawi Puplishing Corporation</publisher><subject>Bacillus subtilis - enzymology ; Bacteria ; Biological and medical sciences ; Cloning, Molecular ; E coli ; Enzyme Stability ; Enzymes ; Escherichia coli - metabolism ; Hydrogen-Ion Concentration ; Medical sciences ; Microbiology ; Pharmacology. Drug treatments ; Proteins ; Recombinant Proteins - chemistry ; Recombinant Proteins - isolation & purification ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Temperature ; Time Factors ; Urate Oxidase - chemistry ; Urate Oxidase - genetics ; Urate Oxidase - isolation & purification</subject><ispartof>BioMed research international, 2010-01, Vol.2010 (2010), p.1-6</ispartof><rights>Copyright © 2010</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2010 Pollyana Pfrimer et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</rights><rights>Copyright © 2010 Pollyana Pfrimer et al. 2010</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c425t-f620d9b3251b7f81029c039cbe98d0b15ef54dd3175ca05bf3638c73172aeb3a3</citedby><cites>FETCH-LOGICAL-c425t-f620d9b3251b7f81029c039cbe98d0b15ef54dd3175ca05bf3638c73172aeb3a3</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/PMC2820260/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2820260/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=25846595$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20168977$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Mowbray, Sherry</contributor><creatorcontrib>Pfrimer, Pollyana</creatorcontrib><creatorcontrib>Moraes, Lídia Maria Pepe de</creatorcontrib><creatorcontrib>Galdino, Alexsandro Sobreira</creatorcontrib><creatorcontrib>Salles, Loise P.</creatorcontrib><creatorcontrib>Reis, Viviane Castelo Branco</creatorcontrib><creatorcontrib>De Marco, Janice Lisboa</creatorcontrib><creatorcontrib>Prates, Maura Vianna</creatorcontrib><creatorcontrib>Bloch, Carlos</creatorcontrib><creatorcontrib>Torres, Fernando Araripe Gonçalves</creatorcontrib><title>Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli</title><title>BioMed research international</title><addtitle>J Biomed Biotechnol</addtitle><description>Urate oxidase (EC 1.7.3.3) is an enzyme involved in purine metabolism which is used in the treatment of gout and as diagnostic reagent for detection of uric acid. In order to produce this enzyme in large quantities for biotechnological purposes, the gene coding for the Bacillus subtilis urate oxidase was cloned and heterologously expressed in Escherichia coli. Time course induction in E. coli showed an induced protein with an apparent molecular mass of ∼60 kDa. Soluble recombinant enzyme was purified in a single-step procedure using Ni-NTA column. The enzyme was purified 2.1-fold with a yield of 56% compared to the crude extract. MALDI-TOF analysis revealed an ion with a mass of 58675 Da which is in agreement with the expected mass of the recombinant protein. The purified enzyme showed an optimal pH and temperature of 8.0 and 37∘C, respectively, and retained 90% of its activity after 72 hours of incubation at −20∘C and 4∘C.</description><subject>Bacillus subtilis - enzymology</subject><subject>Bacteria</subject><subject>Biological and medical sciences</subject><subject>Cloning, Molecular</subject><subject>E coli</subject><subject>Enzyme Stability</subject><subject>Enzymes</subject><subject>Escherichia coli - metabolism</subject><subject>Hydrogen-Ion Concentration</subject><subject>Medical sciences</subject><subject>Microbiology</subject><subject>Pharmacology. Drug treatments</subject><subject>Proteins</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</subject><subject>Temperature</subject><subject>Time Factors</subject><subject>Urate Oxidase - chemistry</subject><subject>Urate Oxidase - genetics</subject><subject>Urate Oxidase - isolation & purification</subject><issn>1110-7243</issn><issn>2314-6133</issn><issn>1110-7251</issn><issn>2314-6141</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>RHX</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqF0Utv1DAQAOAIgWgpnDiDLCQu0KV-xLF9QYLV8pAqtQd6tiaO3UyVOls7gcKvxyXLAidOtjXfPKypqqeMvmFMyhNOGT1pVG2ovlcdMsboSnHJ7u_vtTioHuV8RSlTujEPq4OS0mij1GE1rYcxYrw8JudzwoAOJhzjMYHYkXNIE8JA1j0kcJNP-ONXlIyBvAeHwzBnkud2wgEzuUgweXJ2ix1kTza32-Rz9h3BSDbZ9SXb9QjEjQM-rh4EGLJ_sjuPqosPmy_rT6vTs4-f1-9OV67mclqFhtPOtKJ8plVBM8qNo8K41hvd0ZZJH2TddYIp6YDKNohGaKfKm4NvBYij6u1Sdzu3175zPk4JBrtNeA3pux0B7b-RiL29HL9arjnlDS0FXuwKpPFm9nmyV-OcYpnZatkwRZnhBb1ekEtjzsmHfQNG7d2G7N2G7LKhop__PdPe_l5JAS93ALKDISSIDvMfJ3XdSCOLe7W4HmMH3_A_XZ8t2BfiA-xxrY2smfgJXH2wnw</recordid><startdate>20100101</startdate><enddate>20100101</enddate><creator>Pfrimer, Pollyana</creator><creator>Moraes, Lídia Maria Pepe de</creator><creator>Galdino, Alexsandro Sobreira</creator><creator>Salles, Loise P.</creator><creator>Reis, Viviane Castelo Branco</creator><creator>De Marco, Janice Lisboa</creator><creator>Prates, Maura Vianna</creator><creator>Bloch, Carlos</creator><creator>Torres, Fernando Araripe Gonçalves</creator><general>Hindawi Puplishing Corporation</general><general>Hindawi Publishing Corporation</general><general>Dar al -Nasr -al-Llktruni</general><general>Hindawi Limited</general><scope>ADJCN</scope><scope>AHFXO</scope><scope>RHU</scope><scope>RHW</scope><scope>RHX</scope><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>3V.</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7TK</scope><scope>7U7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>CWDGH</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>5PM</scope></search><sort><creationdate>20100101</creationdate><title>Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli</title><author>Pfrimer, Pollyana ; Moraes, Lídia Maria Pepe de ; Galdino, Alexsandro Sobreira ; Salles, Loise P. ; Reis, Viviane Castelo Branco ; De Marco, Janice Lisboa ; Prates, Maura Vianna ; Bloch, Carlos ; Torres, Fernando Araripe Gonçalves</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c425t-f620d9b3251b7f81029c039cbe98d0b15ef54dd3175ca05bf3638c73172aeb3a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Bacillus subtilis - enzymology</topic><topic>Bacteria</topic><topic>Biological and medical sciences</topic><topic>Cloning, Molecular</topic><topic>E coli</topic><topic>Enzyme Stability</topic><topic>Enzymes</topic><topic>Escherichia coli - metabolism</topic><topic>Hydrogen-Ion Concentration</topic><topic>Medical sciences</topic><topic>Microbiology</topic><topic>Pharmacology. Drug treatments</topic><topic>Proteins</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</topic><topic>Temperature</topic><topic>Time Factors</topic><topic>Urate Oxidase - chemistry</topic><topic>Urate Oxidase - genetics</topic><topic>Urate Oxidase - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pfrimer, Pollyana</creatorcontrib><creatorcontrib>Moraes, Lídia Maria Pepe de</creatorcontrib><creatorcontrib>Galdino, Alexsandro Sobreira</creatorcontrib><creatorcontrib>Salles, Loise P.</creatorcontrib><creatorcontrib>Reis, Viviane Castelo Branco</creatorcontrib><creatorcontrib>De Marco, Janice Lisboa</creatorcontrib><creatorcontrib>Prates, Maura Vianna</creatorcontrib><creatorcontrib>Bloch, Carlos</creatorcontrib><creatorcontrib>Torres, Fernando Araripe Gonçalves</creatorcontrib><collection>الدوريات العلمية والإحصائية - e-Marefa Academic and Statistical Periodicals</collection><collection>معرفة - المحتوى العربي الأكاديمي المتكامل - e-Marefa Academic Complete</collection><collection>Hindawi Publishing Complete</collection><collection>Hindawi Publishing Subscription Journals</collection><collection>Hindawi Publishing Open Access</collection><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>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>Middle East & Africa Database</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>BioMed research international</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pfrimer, Pollyana</au><au>Moraes, Lídia Maria Pepe de</au><au>Galdino, Alexsandro Sobreira</au><au>Salles, Loise P.</au><au>Reis, Viviane Castelo Branco</au><au>De Marco, Janice Lisboa</au><au>Prates, Maura Vianna</au><au>Bloch, Carlos</au><au>Torres, Fernando Araripe Gonçalves</au><au>Mowbray, Sherry</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli</atitle><jtitle>BioMed research international</jtitle><addtitle>J Biomed Biotechnol</addtitle><date>2010-01-01</date><risdate>2010</risdate><volume>2010</volume><issue>2010</issue><spage>1</spage><epage>6</epage><pages>1-6</pages><issn>1110-7243</issn><issn>2314-6133</issn><eissn>1110-7251</eissn><eissn>2314-6141</eissn><abstract>Urate oxidase (EC 1.7.3.3) is an enzyme involved in purine metabolism which is used in the treatment of gout and as diagnostic reagent for detection of uric acid. In order to produce this enzyme in large quantities for biotechnological purposes, the gene coding for the Bacillus subtilis urate oxidase was cloned and heterologously expressed in Escherichia coli. Time course induction in E. coli showed an induced protein with an apparent molecular mass of ∼60 kDa. Soluble recombinant enzyme was purified in a single-step procedure using Ni-NTA column. The enzyme was purified 2.1-fold with a yield of 56% compared to the crude extract. MALDI-TOF analysis revealed an ion with a mass of 58675 Da which is in agreement with the expected mass of the recombinant protein. The purified enzyme showed an optimal pH and temperature of 8.0 and 37∘C, respectively, and retained 90% of its activity after 72 hours of incubation at −20∘C and 4∘C.</abstract><cop>Cairo, Egypt</cop><pub>Hindawi Puplishing Corporation</pub><pmid>20168977</pmid><doi>10.1155/2010/674908</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1110-7243 |
| ispartof | BioMed research international, 2010-01, Vol.2010 (2010), p.1-6 |
| issn | 1110-7243 2314-6133 1110-7251 2314-6141 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2820260 |
| source | MEDLINE; Wiley-Blackwell Open Access Titles; PubMed Central; Alma/SFX Local Collection; PubMed Central Open Access |
| subjects | Bacillus subtilis - enzymology Bacteria Biological and medical sciences Cloning, Molecular E coli Enzyme Stability Enzymes Escherichia coli - metabolism Hydrogen-Ion Concentration Medical sciences Microbiology Pharmacology. Drug treatments Proteins Recombinant Proteins - chemistry Recombinant Proteins - isolation & purification Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Temperature Time Factors Urate Oxidase - chemistry Urate Oxidase - genetics Urate Oxidase - isolation & purification |
| title | Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-12T14%3A05%3A12IST&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=Cloning,%20Purification,%20and%20Partial%20Characterization%20of%20Bacillus%20subtilis%20Urate%20Oxidase%20Expressed%20in%20Escherichia%20coli&rft.jtitle=BioMed%20research%20international&rft.au=Pfrimer,%20Pollyana&rft.date=2010-01-01&rft.volume=2010&rft.issue=2010&rft.spage=1&rft.epage=6&rft.pages=1-6&rft.issn=1110-7243&rft.eissn=1110-7251&rft_id=info:doi/10.1155/2010/674908&rft_dat=%3Cproquest_pubme%3E2287917791%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=856170192&rft_id=info:pmid/20168977&rfr_iscdi=true |