Expression of the cefG gene is limiting for cephalosporin biosynthesis in Acremonium chrysogenum
The conversion of deacetylcephalosporin C to cephalosporin C is inefficient in most Acremonium chrysogenum strains. The cefG gene, which encodes deacetylcephalosporin C acetyltransferase, is expressed very poorly in A. chrysogenum as compared to other genes of the cephalosporin pathway. Introduction...
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| Veröffentlicht in: | Applied microbiology and biotechnology 1997-11, Vol.48 (5), p.606-614 |
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| creator | GUTIERREZ, S VELASCO, J MARCOS, A. T FERNANDEZ, F. J FIERRO, F BARREDO, J. L DIEZ, B MARTIN, J. F |
| description | The conversion of deacetylcephalosporin C to cephalosporin C is inefficient in most Acremonium chrysogenum strains. The cefG gene, which encodes deacetylcephalosporin C acetyltransferase, is expressed very poorly in A. chrysogenum as compared to other genes of the cephalosporin pathway. Introduction of additional copies of the cefG gene with its native promoter (in two different constructions with upstream regions of 1056 bp and 538 bp respectively) did not produce a significant increase of the steady-state level of the cefG transcript. Expression of the cefG gene from the promoters of (i) the glyceraldehyde-3-phosphate dehydrogenase (gpd) gene of Aspergillus nidulans, (ii) the glucoamylase (gla) gene of Aspergillus niger, (iii) the glutamate dehydrogenase (gdh) and (iv) the isopenicillin N synthase (pcbC) genes of Penicillium chrysogenum, led to very high steady-state levels of cefG transcript and to increased deacetylcephalosporin-C acetyltransferase protein concentration (as shown by immunoblotting) and enzyme activity in the transformants. Southern analysis showed that integration of the new constructions occurred at sites different from that of the endogenous cefG gene. Cephalosporin production was increased two- to threefold in A. chrysogenum C10 transformed with constructions in which the cefG gene was expressed from the gdh or gpd promoters as a result of a more efficient acetylation of deacetylcephalosporin C. |
| doi_str_mv | 10.1007/s002530051103 |
| format | Article |
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T ; FERNANDEZ, F. J ; FIERRO, F ; BARREDO, J. L ; DIEZ, B ; MARTIN, J. F</creator><creatorcontrib>GUTIERREZ, S ; VELASCO, J ; MARCOS, A. T ; FERNANDEZ, F. J ; FIERRO, F ; BARREDO, J. L ; DIEZ, B ; MARTIN, J. F</creatorcontrib><description>The conversion of deacetylcephalosporin C to cephalosporin C is inefficient in most Acremonium chrysogenum strains. The cefG gene, which encodes deacetylcephalosporin C acetyltransferase, is expressed very poorly in A. chrysogenum as compared to other genes of the cephalosporin pathway. Introduction of additional copies of the cefG gene with its native promoter (in two different constructions with upstream regions of 1056 bp and 538 bp respectively) did not produce a significant increase of the steady-state level of the cefG transcript. Expression of the cefG gene from the promoters of (i) the glyceraldehyde-3-phosphate dehydrogenase (gpd) gene of Aspergillus nidulans, (ii) the glucoamylase (gla) gene of Aspergillus niger, (iii) the glutamate dehydrogenase (gdh) and (iv) the isopenicillin N synthase (pcbC) genes of Penicillium chrysogenum, led to very high steady-state levels of cefG transcript and to increased deacetylcephalosporin-C acetyltransferase protein concentration (as shown by immunoblotting) and enzyme activity in the transformants. Southern analysis showed that integration of the new constructions occurred at sites different from that of the endogenous cefG gene. Cephalosporin production was increased two- to threefold in A. chrysogenum C10 transformed with constructions in which the cefG gene was expressed from the gdh or gpd promoters as a result of a more efficient acetylation of deacetylcephalosporin C.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s002530051103</identifier><identifier>PMID: 9421924</identifier><identifier>CODEN: AMBIDG</identifier><language>eng</language><publisher>Berlin: Springer</publisher><subject>Acetyltransferases - genetics ; Acetyltransferases - immunology ; Acetyltransferases - metabolism ; Acremonium - genetics ; Acremonium - metabolism ; Acremonium chrysogenum ; Aspergillus nidulans ; Aspergillus niger ; Beta lactam antibiotics ; Biological and medical sciences ; Biosynthesis ; Biotechnology ; Cephalosporins - biosynthesis ; Chromosome Mapping ; Cloning, Molecular ; Dehydrogenase ; Dehydrogenases ; DNA, Fungal - analysis ; DNA, Fungal - genetics ; Enzymatic activity ; Fundamental and applied biological sciences. Psychology ; Gene Amplification ; Gene Dosage ; Gene expression ; Gene Expression Regulation, Fungal ; Genes ; Genetic aspects ; Genetic engineering ; Genetic technics ; Glucan 1,4-alpha-Glucosidase - genetics ; Glyceraldehyde-3-Phosphate Dehydrogenases - genetics ; Hydro-Lyases - genetics ; Immunoblotting ; Methods. Procedures. Technologies ; Microbial genetics ; Modification of gene expression level ; Oxidoreductases - genetics ; Penicillium chrysogenum ; Pharmaceutical chemistry ; Physiological aspects ; Plasmids ; Promoter Regions, Genetic ; Recombination, Genetic ; Transcription, Genetic</subject><ispartof>Applied microbiology and biotechnology, 1997-11, Vol.48 (5), p.606-614</ispartof><rights>1998 INIST-CNRS</rights><rights>COPYRIGHT 1997 Springer</rights><rights>Springer-Verlag Berlin Heidelberg 1997</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c473t-d4290b5e47a4e03cd3e7070b9c4113a0bf4ad8424abba7478051a656037548333</citedby></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=2100374$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9421924$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>GUTIERREZ, S</creatorcontrib><creatorcontrib>VELASCO, J</creatorcontrib><creatorcontrib>MARCOS, A. T</creatorcontrib><creatorcontrib>FERNANDEZ, F. J</creatorcontrib><creatorcontrib>FIERRO, F</creatorcontrib><creatorcontrib>BARREDO, J. L</creatorcontrib><creatorcontrib>DIEZ, B</creatorcontrib><creatorcontrib>MARTIN, J. F</creatorcontrib><title>Expression of the cefG gene is limiting for cephalosporin biosynthesis in Acremonium chrysogenum</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><description>The conversion of deacetylcephalosporin C to cephalosporin C is inefficient in most Acremonium chrysogenum strains. The cefG gene, which encodes deacetylcephalosporin C acetyltransferase, is expressed very poorly in A. chrysogenum as compared to other genes of the cephalosporin pathway. Introduction of additional copies of the cefG gene with its native promoter (in two different constructions with upstream regions of 1056 bp and 538 bp respectively) did not produce a significant increase of the steady-state level of the cefG transcript. Expression of the cefG gene from the promoters of (i) the glyceraldehyde-3-phosphate dehydrogenase (gpd) gene of Aspergillus nidulans, (ii) the glucoamylase (gla) gene of Aspergillus niger, (iii) the glutamate dehydrogenase (gdh) and (iv) the isopenicillin N synthase (pcbC) genes of Penicillium chrysogenum, led to very high steady-state levels of cefG transcript and to increased deacetylcephalosporin-C acetyltransferase protein concentration (as shown by immunoblotting) and enzyme activity in the transformants. Southern analysis showed that integration of the new constructions occurred at sites different from that of the endogenous cefG gene. Cephalosporin production was increased two- to threefold in A. chrysogenum C10 transformed with constructions in which the cefG gene was expressed from the gdh or gpd promoters as a result of a more efficient acetylation of deacetylcephalosporin C.</description><subject>Acetyltransferases - genetics</subject><subject>Acetyltransferases - immunology</subject><subject>Acetyltransferases - metabolism</subject><subject>Acremonium - genetics</subject><subject>Acremonium - metabolism</subject><subject>Acremonium chrysogenum</subject><subject>Aspergillus nidulans</subject><subject>Aspergillus niger</subject><subject>Beta lactam antibiotics</subject><subject>Biological and medical sciences</subject><subject>Biosynthesis</subject><subject>Biotechnology</subject><subject>Cephalosporins - biosynthesis</subject><subject>Chromosome Mapping</subject><subject>Cloning, Molecular</subject><subject>Dehydrogenase</subject><subject>Dehydrogenases</subject><subject>DNA, Fungal - analysis</subject><subject>DNA, Fungal - genetics</subject><subject>Enzymatic activity</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Amplification</subject><subject>Gene Dosage</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Fungal</subject><subject>Genes</subject><subject>Genetic aspects</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Glucan 1,4-alpha-Glucosidase - genetics</subject><subject>Glyceraldehyde-3-Phosphate Dehydrogenases - genetics</subject><subject>Hydro-Lyases - genetics</subject><subject>Immunoblotting</subject><subject>Methods. Procedures. Technologies</subject><subject>Microbial genetics</subject><subject>Modification of gene expression level</subject><subject>Oxidoreductases - genetics</subject><subject>Penicillium chrysogenum</subject><subject>Pharmaceutical chemistry</subject><subject>Physiological aspects</subject><subject>Plasmids</subject><subject>Promoter Regions, Genetic</subject><subject>Recombination, Genetic</subject><subject>Transcription, Genetic</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpd0d-L1DAQB_AgyrmuPvooFE8QH6qTZtK0j8txngcHgj-eY5pNd3O0Sc20cPvfm2OXg_UphPnkm0mGsbccPnMA9YUAKikAJOcgnrEVR1GVUHN8zlbAlSyVbJuX7BXRPQCvmrq-YBctVrytcMX-XD9MyRH5GIrYF_PeFdb1N8XOBVd4KgY_-tmHXdHHlCvT3gyRpph8KDof6RDyCcou7zc2uTEGv4yF3acDxZyxjK_Zi94M5N6c1jX7_fX619W38u77ze3V5q60qMRcbrFqoZMOlUEHwm6FU6Cgay1yLgx0PZptgxWarjMKVZPfa2pZg1ASGyHEmn085k4p_l0czXr0ZN0wmODiQroFxFao_Edr9v4_eR-XFHJzummwlvkSmdHlEe3M4LQPfZyTsY-ReiOA8wYEYFafzpSNYXYP884sRPr2549zWx6tTZEouV5PyY8mHTQH_ThJfTbJ7N-d2ly60W2f9Gl0uf7hVDdkzdAnE6ynJ1blSKFQ_APayKId</recordid><startdate>19971101</startdate><enddate>19971101</enddate><creator>GUTIERREZ, S</creator><creator>VELASCO, J</creator><creator>MARCOS, A. 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T</au><au>FERNANDEZ, F. J</au><au>FIERRO, F</au><au>BARREDO, J. L</au><au>DIEZ, B</au><au>MARTIN, J. F</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of the cefG gene is limiting for cephalosporin biosynthesis in Acremonium chrysogenum</atitle><jtitle>Applied microbiology and biotechnology</jtitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>1997-11-01</date><risdate>1997</risdate><volume>48</volume><issue>5</issue><spage>606</spage><epage>614</epage><pages>606-614</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><coden>AMBIDG</coden><abstract>The conversion of deacetylcephalosporin C to cephalosporin C is inefficient in most Acremonium chrysogenum strains. The cefG gene, which encodes deacetylcephalosporin C acetyltransferase, is expressed very poorly in A. chrysogenum as compared to other genes of the cephalosporin pathway. Introduction of additional copies of the cefG gene with its native promoter (in two different constructions with upstream regions of 1056 bp and 538 bp respectively) did not produce a significant increase of the steady-state level of the cefG transcript. Expression of the cefG gene from the promoters of (i) the glyceraldehyde-3-phosphate dehydrogenase (gpd) gene of Aspergillus nidulans, (ii) the glucoamylase (gla) gene of Aspergillus niger, (iii) the glutamate dehydrogenase (gdh) and (iv) the isopenicillin N synthase (pcbC) genes of Penicillium chrysogenum, led to very high steady-state levels of cefG transcript and to increased deacetylcephalosporin-C acetyltransferase protein concentration (as shown by immunoblotting) and enzyme activity in the transformants. Southern analysis showed that integration of the new constructions occurred at sites different from that of the endogenous cefG gene. Cephalosporin production was increased two- to threefold in A. chrysogenum C10 transformed with constructions in which the cefG gene was expressed from the gdh or gpd promoters as a result of a more efficient acetylation of deacetylcephalosporin C.</abstract><cop>Berlin</cop><pub>Springer</pub><pmid>9421924</pmid><doi>10.1007/s002530051103</doi><tpages>9</tpages></addata></record> |
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| ispartof | Applied microbiology and biotechnology, 1997-11, Vol.48 (5), p.606-614 |
| issn | 0175-7598 1432-0614 |
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| source | MEDLINE; SpringerLink Journals |
| subjects | Acetyltransferases - genetics Acetyltransferases - immunology Acetyltransferases - metabolism Acremonium - genetics Acremonium - metabolism Acremonium chrysogenum Aspergillus nidulans Aspergillus niger Beta lactam antibiotics Biological and medical sciences Biosynthesis Biotechnology Cephalosporins - biosynthesis Chromosome Mapping Cloning, Molecular Dehydrogenase Dehydrogenases DNA, Fungal - analysis DNA, Fungal - genetics Enzymatic activity Fundamental and applied biological sciences. Psychology Gene Amplification Gene Dosage Gene expression Gene Expression Regulation, Fungal Genes Genetic aspects Genetic engineering Genetic technics Glucan 1,4-alpha-Glucosidase - genetics Glyceraldehyde-3-Phosphate Dehydrogenases - genetics Hydro-Lyases - genetics Immunoblotting Methods. Procedures. Technologies Microbial genetics Modification of gene expression level Oxidoreductases - genetics Penicillium chrysogenum Pharmaceutical chemistry Physiological aspects Plasmids Promoter Regions, Genetic Recombination, Genetic Transcription, Genetic |
| title | Expression of the cefG gene is limiting for cephalosporin biosynthesis in Acremonium chrysogenum |
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