The clavulanic acid biosynthetic cluster of Streptomyces clavuligerus: genetic organization of the region upstream of the car gene
Área de Biotecnología, Antibióticos SA, Avenida de Antibióticos 59-61, 24009 León, Spain 1 Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain 2 Author for correspondence: José L. Barredo. Tel: +34 987 895826. Fax: +34 987 895986. e-mail: jbarredo{at}antibioticos.it T...
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| Veröffentlicht in: | Microbiology (Society for General Microbiology) 2002-05, Vol.148 (5), p.1427-1438 |
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| creator | Mellado, Encarnacion Lorenzana, Luis Miguel Rodriguez-Saiz, Marta Diez, Bruno Liras, Paloma Barredo, Jose Luis |
| description | Área de Biotecnología, Antibióticos SA, Avenida de Antibióticos 59-61, 24009 León, Spain 1
Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain 2
Author for correspondence: José L. Barredo. Tel: +34 987 895826. Fax: +34 987 895986. e-mail: jbarredo{at}antibioticos.it
The genetic organization of the region upstream of the car gene of the clavulanic acid biosynthetic gene cluster of Streptomyces clavuligerus has been determined. Sequence analysis of a 12·1 kb region revealed the presence of 10 ORFs whose putative functions, according to database searches, are discussed. Three co-transcriptional units are proposed: ORF1011, ORF1213 and ORF15161718. Potential transcriptional terminators were identified downstream of ORF11 ( fd ) and ORF15. Targeted disruption of ORF10 ( cyp ) gave rise to transformants unable to produce clavulanic acid, but with a considerably higher production of cephamycin C. Transformants inactivated at ORF14 had a remarkably lower production of clavulanic acid and similar production of cephamycin C. Significant improvements of clavulanic acid production, associated with a drop in cephamycin C biosynthesis, were obtained with transformants of S. clavuligerus harbouring multiple copies of plasmids carrying different constructions from the ORF1014 region. This information can be used to guide strain improvement programs, blending random mutagenesis and molecular cloning, to optimize the yield of clavulanic acid.
Keywords: antibiotic biosynthesis, gene cluster, cytochrome P450, ferredoxin, PBP Abbreviations: CA, clavulanic acid; PBP, penicillin-binding protein
The GenBank accession number for the 12162 bp sequence reported in this paper is AY034175 . |
| doi_str_mv | 10.1099/00221287-148-5-1427 |
| format | Article |
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Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain 2
Author for correspondence: José L. Barredo. Tel: +34 987 895826. Fax: +34 987 895986. e-mail: jbarredo{at}antibioticos.it
The genetic organization of the region upstream of the car gene of the clavulanic acid biosynthetic gene cluster of Streptomyces clavuligerus has been determined. Sequence analysis of a 12·1 kb region revealed the presence of 10 ORFs whose putative functions, according to database searches, are discussed. Three co-transcriptional units are proposed: ORF1011, ORF1213 and ORF15161718. Potential transcriptional terminators were identified downstream of ORF11 ( fd ) and ORF15. Targeted disruption of ORF10 ( cyp ) gave rise to transformants unable to produce clavulanic acid, but with a considerably higher production of cephamycin C. Transformants inactivated at ORF14 had a remarkably lower production of clavulanic acid and similar production of cephamycin C. Significant improvements of clavulanic acid production, associated with a drop in cephamycin C biosynthesis, were obtained with transformants of S. clavuligerus harbouring multiple copies of plasmids carrying different constructions from the ORF1014 region. This information can be used to guide strain improvement programs, blending random mutagenesis and molecular cloning, to optimize the yield of clavulanic acid.
Keywords: antibiotic biosynthesis, gene cluster, cytochrome P450, ferredoxin, PBP Abbreviations: CA, clavulanic acid; PBP, penicillin-binding protein
The GenBank accession number for the 12162 bp sequence reported in this paper is AY034175 .</description><identifier>ISSN: 1350-0872</identifier><identifier>EISSN: 1465-2080</identifier><identifier>DOI: 10.1099/00221287-148-5-1427</identifier><identifier>PMID: 11988517</identifier><language>eng</language><publisher>Reading: Soc General Microbiol</publisher><subject>Bacteriology ; Base Sequence ; Biological and medical sciences ; car gene ; cephamycin C ; Cephamycins - biosynthesis ; clavulanic acid ; Clavulanic Acid - biosynthesis ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Gene Expression Regulation, Bacterial ; Gene Order ; Genes, Bacterial - genetics ; Genetics ; Genomic Library ; Microbiology ; Molecular Sequence Data ; Open Reading Frames - genetics ; Plasmids - genetics ; Sequence Analysis, DNA ; Streptomyces - genetics ; Streptomyces - growth & development ; Streptomyces - metabolism ; Streptomyces clavuligerus ; Transcription, Genetic - genetics ; Transformation, Bacterial</subject><ispartof>Microbiology (Society for General Microbiology), 2002-05, Vol.148 (5), p.1427-1438</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14283342$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11988517$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mellado, Encarnacion</creatorcontrib><creatorcontrib>Lorenzana, Luis Miguel</creatorcontrib><creatorcontrib>Rodriguez-Saiz, Marta</creatorcontrib><creatorcontrib>Diez, Bruno</creatorcontrib><creatorcontrib>Liras, Paloma</creatorcontrib><creatorcontrib>Barredo, Jose Luis</creatorcontrib><title>The clavulanic acid biosynthetic cluster of Streptomyces clavuligerus: genetic organization of the region upstream of the car gene</title><title>Microbiology (Society for General Microbiology)</title><addtitle>Microbiology</addtitle><description>Área de Biotecnología, Antibióticos SA, Avenida de Antibióticos 59-61, 24009 León, Spain 1
Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain 2
Author for correspondence: José L. Barredo. Tel: +34 987 895826. Fax: +34 987 895986. e-mail: jbarredo{at}antibioticos.it
The genetic organization of the region upstream of the car gene of the clavulanic acid biosynthetic gene cluster of Streptomyces clavuligerus has been determined. Sequence analysis of a 12·1 kb region revealed the presence of 10 ORFs whose putative functions, according to database searches, are discussed. Three co-transcriptional units are proposed: ORF1011, ORF1213 and ORF15161718. Potential transcriptional terminators were identified downstream of ORF11 ( fd ) and ORF15. Targeted disruption of ORF10 ( cyp ) gave rise to transformants unable to produce clavulanic acid, but with a considerably higher production of cephamycin C. Transformants inactivated at ORF14 had a remarkably lower production of clavulanic acid and similar production of cephamycin C. Significant improvements of clavulanic acid production, associated with a drop in cephamycin C biosynthesis, were obtained with transformants of S. clavuligerus harbouring multiple copies of plasmids carrying different constructions from the ORF1014 region. This information can be used to guide strain improvement programs, blending random mutagenesis and molecular cloning, to optimize the yield of clavulanic acid.
Keywords: antibiotic biosynthesis, gene cluster, cytochrome P450, ferredoxin, PBP Abbreviations: CA, clavulanic acid; PBP, penicillin-binding protein
The GenBank accession number for the 12162 bp sequence reported in this paper is AY034175 .</description><subject>Bacteriology</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>car gene</subject><subject>cephamycin C</subject><subject>Cephamycins - biosynthesis</subject><subject>clavulanic acid</subject><subject>Clavulanic Acid - biosynthesis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Gene Order</subject><subject>Genes, Bacterial - genetics</subject><subject>Genetics</subject><subject>Genomic Library</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Open Reading Frames - genetics</subject><subject>Plasmids - genetics</subject><subject>Sequence Analysis, DNA</subject><subject>Streptomyces - genetics</subject><subject>Streptomyces - growth & development</subject><subject>Streptomyces - metabolism</subject><subject>Streptomyces clavuligerus</subject><subject>Transcription, Genetic - genetics</subject><subject>Transformation, Bacterial</subject><issn>1350-0872</issn><issn>1465-2080</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kcuO1TAMhiMEYi7wBEioG9CwKDhOQpPZoRE3aSQWDOsqTd02qJdD0oIOS54cn5kzYscmie3v_yPbQjyT8FqCc28AECXaqpTaloZPrB6IU6nfmhLBwkN-KwMl2ApPxFnO3wG4CPKxOJHSWWtkdSr-3AxUhNH_3EY_x1D4ENuiiUvez-tAK2fCuOWVUrF0xdc10W5dpn2gfBTFntKWL4ue5lt6ST37_PZrXOaDhE2KRP0h2naZ9X66TwefbmVPxKPOj5meHu9z8e3D-5urT-X1l4-fr95dl4NSZi1dqFrjnMYGq-CAlHcW0Letc4o6amWjXdMSGG4SPRgEr5pGddhwwSurzsXFne8uLT82yms9xRxo5MZp2XItHaCUCrVi9OX_UatBGzQMPj-CWzNRW-9SnHza1_cDZuDFEfA5-LFLfg4x_-M0WqU0MvfqjhtiP_yKiWoezRRDWngZ_HvgJdfmwFfqL_d9mtU</recordid><startdate>20020501</startdate><enddate>20020501</enddate><creator>Mellado, Encarnacion</creator><creator>Lorenzana, Luis Miguel</creator><creator>Rodriguez-Saiz, Marta</creator><creator>Diez, Bruno</creator><creator>Liras, Paloma</creator><creator>Barredo, Jose Luis</creator><general>Soc General Microbiol</general><general>Society for General Microbiology</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7QL</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>20020501</creationdate><title>The clavulanic acid biosynthetic cluster of Streptomyces clavuligerus: genetic organization of the region upstream of the car gene</title><author>Mellado, Encarnacion ; Lorenzana, Luis Miguel ; Rodriguez-Saiz, Marta ; Diez, Bruno ; Liras, Paloma ; Barredo, Jose Luis</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-h335t-9c7d59942b27c90e3a9802add993efed1b49bde051462a0520a3bb3f2bb49a383</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Bacteriology</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>car gene</topic><topic>cephamycin C</topic><topic>Cephamycins - biosynthesis</topic><topic>clavulanic acid</topic><topic>Clavulanic Acid - biosynthesis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Gene Expression Regulation, Bacterial</topic><topic>Gene Order</topic><topic>Genes, Bacterial - genetics</topic><topic>Genetics</topic><topic>Genomic Library</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>Open Reading Frames - genetics</topic><topic>Plasmids - genetics</topic><topic>Sequence Analysis, DNA</topic><topic>Streptomyces - genetics</topic><topic>Streptomyces - growth & development</topic><topic>Streptomyces - metabolism</topic><topic>Streptomyces clavuligerus</topic><topic>Transcription, Genetic - genetics</topic><topic>Transformation, Bacterial</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mellado, Encarnacion</creatorcontrib><creatorcontrib>Lorenzana, Luis Miguel</creatorcontrib><creatorcontrib>Rodriguez-Saiz, Marta</creatorcontrib><creatorcontrib>Diez, Bruno</creatorcontrib><creatorcontrib>Liras, Paloma</creatorcontrib><creatorcontrib>Barredo, Jose Luis</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Microbiology (Society for General Microbiology)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mellado, Encarnacion</au><au>Lorenzana, Luis Miguel</au><au>Rodriguez-Saiz, Marta</au><au>Diez, Bruno</au><au>Liras, Paloma</au><au>Barredo, Jose Luis</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The clavulanic acid biosynthetic cluster of Streptomyces clavuligerus: genetic organization of the region upstream of the car gene</atitle><jtitle>Microbiology (Society for General Microbiology)</jtitle><addtitle>Microbiology</addtitle><date>2002-05-01</date><risdate>2002</risdate><volume>148</volume><issue>5</issue><spage>1427</spage><epage>1438</epage><pages>1427-1438</pages><issn>1350-0872</issn><eissn>1465-2080</eissn><abstract>Área de Biotecnología, Antibióticos SA, Avenida de Antibióticos 59-61, 24009 León, Spain 1
Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain 2
Author for correspondence: José L. Barredo. Tel: +34 987 895826. Fax: +34 987 895986. e-mail: jbarredo{at}antibioticos.it
The genetic organization of the region upstream of the car gene of the clavulanic acid biosynthetic gene cluster of Streptomyces clavuligerus has been determined. Sequence analysis of a 12·1 kb region revealed the presence of 10 ORFs whose putative functions, according to database searches, are discussed. Three co-transcriptional units are proposed: ORF1011, ORF1213 and ORF15161718. Potential transcriptional terminators were identified downstream of ORF11 ( fd ) and ORF15. Targeted disruption of ORF10 ( cyp ) gave rise to transformants unable to produce clavulanic acid, but with a considerably higher production of cephamycin C. Transformants inactivated at ORF14 had a remarkably lower production of clavulanic acid and similar production of cephamycin C. Significant improvements of clavulanic acid production, associated with a drop in cephamycin C biosynthesis, were obtained with transformants of S. clavuligerus harbouring multiple copies of plasmids carrying different constructions from the ORF1014 region. This information can be used to guide strain improvement programs, blending random mutagenesis and molecular cloning, to optimize the yield of clavulanic acid.
Keywords: antibiotic biosynthesis, gene cluster, cytochrome P450, ferredoxin, PBP Abbreviations: CA, clavulanic acid; PBP, penicillin-binding protein
The GenBank accession number for the 12162 bp sequence reported in this paper is AY034175 .</abstract><cop>Reading</cop><pub>Soc General Microbiol</pub><pmid>11988517</pmid><doi>10.1099/00221287-148-5-1427</doi><tpages>12</tpages></addata></record> |
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| ispartof | Microbiology (Society for General Microbiology), 2002-05, Vol.148 (5), p.1427-1438 |
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| subjects | Bacteriology Base Sequence Biological and medical sciences car gene cephamycin C Cephamycins - biosynthesis clavulanic acid Clavulanic Acid - biosynthesis Fundamental and applied biological sciences. Psychology Gene Expression Gene Expression Regulation, Bacterial Gene Order Genes, Bacterial - genetics Genetics Genomic Library Microbiology Molecular Sequence Data Open Reading Frames - genetics Plasmids - genetics Sequence Analysis, DNA Streptomyces - genetics Streptomyces - growth & development Streptomyces - metabolism Streptomyces clavuligerus Transcription, Genetic - genetics Transformation, Bacterial |
| title | The clavulanic acid biosynthetic cluster of Streptomyces clavuligerus: genetic organization of the region upstream of the car gene |
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