Isolation and molecular characterization of high-performance cellobiose-fermenting spontaneous mutants of ethanologenic Escherichia coli KO11 containing the Klebsiella oxytoca casAB operon
Escherichia coli KO11 was previously constructed to produce ethanol from acid hydrolysates of hemicellulose (pentoses and hexoses) by the chromosomal integration of Zymomonas mobilis genes encoding pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB). Klebsiella oxytoca P2 was constructed i...
Gespeichert in:
| Veröffentlicht in: | Applied and Environmental Microbiology 1997-12, Vol.63 (12), p.4633-4637 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 4637 |
|---|---|
| container_issue | 12 |
| container_start_page | 4633 |
| container_title | Applied and Environmental Microbiology |
| container_volume | 63 |
| creator | Moniruzzaman, M Lai, X York, S.W Ingram, L.O |
| description | Escherichia coli KO11 was previously constructed to produce ethanol from acid hydrolysates of hemicellulose (pentoses and hexoses) by the chromosomal integration of Zymomonas mobilis genes encoding pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB). Klebsiella oxytoca P2 was constructed in an analogous fashion for the simultaneous saccharification and fermentation of cellulose and contains PTS enzymes for cellobiose. In this study, KO11 was further engineered for the fermentation of cellulose by adding the K. oxytoca casAB genes encoding Enzyme II cellobiose and phosphobetaglucosidase. Although the two K. oxytoca genes were well expressed in cloning hosts such as DH5 alpha both were expressed poorly in E. coli KO11, a derivative of E. coli B. Spontaneous mutants which exhibited more than 15-fold-higher specific activities for cellobiose metabolism were isolated. The mutations of these mutants resided in the plasmid rather than the host. Three mutants were characterized by sequence analysis. All contained similar internal deletions which eliminated the casAB promoter and operator regions and placed the lacZ Shine-Dalgarno region immediately upstream from the casA Shine-Dalgarno region. KO11 harboring mutant plasmids (pLOI1908, pLOI1909, or pLOI1910) rapidly fermented cellobiose to ethanol, and the yield was more than 90% of the theoretical yield. Two of these strains were used with commercial cellulase to ferment mixed-waste office paper to ethanol |
| doi_str_mv | 10.1128/aem.63.12.4633-4637.1997 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_168784</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>79475072</sourcerecordid><originalsourceid>FETCH-LOGICAL-c702t-4b7b5df9c35c14dfa832029ff4f0ea15830112d989032f5bc33c7bb39924651e3</originalsourceid><addsrcrecordid>eNqFUstu1DAUjRCoDAOfgGQhxC6DH3FsL1iUqkDVSl1A19aNx564SuzBzgDl2_g4HM1oCt3Ui-sr3XPu4-hUFSJ4RQiV78GOq5atCF01LWN1CWJFlBJPqgXBStacsfZptcBYqZrSBj-vXuR8izFucCtPqhNVfibxovpzkeMAk48BQVijMQ7W7AZIyPSQwEw2-d_7cnSo95u-3trkYhohGIuMHYbY-Zht7WwabZh82KC8jWGCYOMuo3FXsinPbDv1EOIQNzZ4g86z6Utz03tAJg4eXV4TUrLC9GHuMvUWXQ62y74MARR_3U3RFCzk048oli1ieFk9czBk--rwL6ubT-ffzr7UV9efL85Or2ojMJ3qphMdXztlGDekWTuQjGKqnGsctkC4ZLiIulZSYUYd7wxjRnQdU4o2LSeWLasP-77bXTfatSl3Jhj0NvkR0p2O4PX_leB7vYk_NGmlkE3hvzvwU_y-s3nSo8-zdnuRtFCN4FjQR4FUsAZLxR8FkpZhwcpbVm8eAG_jLoWilqaYFxtwJQpI7kEmxZyTdcfTCNaz33Txm26ZJlTPfpuD0LPfCvX1v9IciQeDlfrbQx2ygcGl4hufjzBKSMNUe7_mbLGfPlkNeXww9X6Wg6hhk0qfm6_zFrgVknP2F3GU-Eg</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>205940597</pqid></control><display><type>article</type><title>Isolation and molecular characterization of high-performance cellobiose-fermenting spontaneous mutants of ethanologenic Escherichia coli KO11 containing the Klebsiella oxytoca casAB operon</title><source>Open Access: PubMed Central</source><source>MEDLINE</source><source>American Society for Microbiology Journals</source><source>Alma/SFX Local Collection</source><creator>Moniruzzaman, M ; Lai, X ; York, S.W ; Ingram, L.O</creator><creatorcontrib>Moniruzzaman, M ; Lai, X ; York, S.W ; Ingram, L.O</creatorcontrib><description>Escherichia coli KO11 was previously constructed to produce ethanol from acid hydrolysates of hemicellulose (pentoses and hexoses) by the chromosomal integration of Zymomonas mobilis genes encoding pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB). Klebsiella oxytoca P2 was constructed in an analogous fashion for the simultaneous saccharification and fermentation of cellulose and contains PTS enzymes for cellobiose. In this study, KO11 was further engineered for the fermentation of cellulose by adding the K. oxytoca casAB genes encoding Enzyme II cellobiose and phosphobetaglucosidase. Although the two K. oxytoca genes were well expressed in cloning hosts such as DH5 alpha both were expressed poorly in E. coli KO11, a derivative of E. coli B. Spontaneous mutants which exhibited more than 15-fold-higher specific activities for cellobiose metabolism were isolated. The mutations of these mutants resided in the plasmid rather than the host. Three mutants were characterized by sequence analysis. All contained similar internal deletions which eliminated the casAB promoter and operator regions and placed the lacZ Shine-Dalgarno region immediately upstream from the casA Shine-Dalgarno region. KO11 harboring mutant plasmids (pLOI1908, pLOI1909, or pLOI1910) rapidly fermented cellobiose to ethanol, and the yield was more than 90% of the theoretical yield. Two of these strains were used with commercial cellulase to ferment mixed-waste office paper to ethanol</description><identifier>ISSN: 0099-2240</identifier><identifier>EISSN: 1098-5336</identifier><identifier>DOI: 10.1128/aem.63.12.4633-4637.1997</identifier><identifier>PMID: 9406380</identifier><identifier>CODEN: AEMIDF</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>Bacteria ; Base Sequence ; Biological and medical sciences ; Biology of microorganisms of confirmed or potential industrial interest ; Biotechnology ; Cellobiose - metabolism ; DECHET ; DESECHOS ; DNA Primers - genetics ; DNA, Bacterial - genetics ; ESCHERICHIA COLI ; Escherichia coli - genetics ; Escherichia coli - isolation & purification ; Escherichia coli - metabolism ; ETANOL ; ETHANOL ; Ethanol - metabolism ; ETHANOL PRODUCTION ; FERMENTACION ; FERMENTATION ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Genes, Bacterial ; Genetics ; KLEBSIELLA ; Klebsiella - genetics ; Microbiology ; Mission oriented research ; Molecular Sequence Data ; MUTANT ; MUTANTES ; MUTANTS ; Mutation ; Operon ; PAPEL ; PAPIER ; WASTES</subject><ispartof>Applied and Environmental Microbiology, 1997-12, Vol.63 (12), p.4633-4637</ispartof><rights>1998 INIST-CNRS</rights><rights>Copyright American Society for Microbiology Dec 1997</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c702t-4b7b5df9c35c14dfa832029ff4f0ea15830112d989032f5bc33c7bb39924651e3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC168784/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC168784/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,3175,3176,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2114396$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9406380$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Moniruzzaman, M</creatorcontrib><creatorcontrib>Lai, X</creatorcontrib><creatorcontrib>York, S.W</creatorcontrib><creatorcontrib>Ingram, L.O</creatorcontrib><title>Isolation and molecular characterization of high-performance cellobiose-fermenting spontaneous mutants of ethanologenic Escherichia coli KO11 containing the Klebsiella oxytoca casAB operon</title><title>Applied and Environmental Microbiology</title><addtitle>Appl Environ Microbiol</addtitle><description>Escherichia coli KO11 was previously constructed to produce ethanol from acid hydrolysates of hemicellulose (pentoses and hexoses) by the chromosomal integration of Zymomonas mobilis genes encoding pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB). Klebsiella oxytoca P2 was constructed in an analogous fashion for the simultaneous saccharification and fermentation of cellulose and contains PTS enzymes for cellobiose. In this study, KO11 was further engineered for the fermentation of cellulose by adding the K. oxytoca casAB genes encoding Enzyme II cellobiose and phosphobetaglucosidase. Although the two K. oxytoca genes were well expressed in cloning hosts such as DH5 alpha both were expressed poorly in E. coli KO11, a derivative of E. coli B. Spontaneous mutants which exhibited more than 15-fold-higher specific activities for cellobiose metabolism were isolated. The mutations of these mutants resided in the plasmid rather than the host. Three mutants were characterized by sequence analysis. All contained similar internal deletions which eliminated the casAB promoter and operator regions and placed the lacZ Shine-Dalgarno region immediately upstream from the casA Shine-Dalgarno region. KO11 harboring mutant plasmids (pLOI1908, pLOI1909, or pLOI1910) rapidly fermented cellobiose to ethanol, and the yield was more than 90% of the theoretical yield. Two of these strains were used with commercial cellulase to ferment mixed-waste office paper to ethanol</description><subject>Bacteria</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biology of microorganisms of confirmed or potential industrial interest</subject><subject>Biotechnology</subject><subject>Cellobiose - metabolism</subject><subject>DECHET</subject><subject>DESECHOS</subject><subject>DNA Primers - genetics</subject><subject>DNA, Bacterial - genetics</subject><subject>ESCHERICHIA COLI</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - isolation & purification</subject><subject>Escherichia coli - metabolism</subject><subject>ETANOL</subject><subject>ETHANOL</subject><subject>Ethanol - metabolism</subject><subject>ETHANOL PRODUCTION</subject><subject>FERMENTACION</subject><subject>FERMENTATION</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genes, Bacterial</subject><subject>Genetics</subject><subject>KLEBSIELLA</subject><subject>Klebsiella - genetics</subject><subject>Microbiology</subject><subject>Mission oriented research</subject><subject>Molecular Sequence Data</subject><subject>MUTANT</subject><subject>MUTANTES</subject><subject>MUTANTS</subject><subject>Mutation</subject><subject>Operon</subject><subject>PAPEL</subject><subject>PAPIER</subject><subject>WASTES</subject><issn>0099-2240</issn><issn>1098-5336</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFUstu1DAUjRCoDAOfgGQhxC6DH3FsL1iUqkDVSl1A19aNx564SuzBzgDl2_g4HM1oCt3Ui-sr3XPu4-hUFSJ4RQiV78GOq5atCF01LWN1CWJFlBJPqgXBStacsfZptcBYqZrSBj-vXuR8izFucCtPqhNVfibxovpzkeMAk48BQVijMQ7W7AZIyPSQwEw2-d_7cnSo95u-3trkYhohGIuMHYbY-Zht7WwabZh82KC8jWGCYOMuo3FXsinPbDv1EOIQNzZ4g86z6Utz03tAJg4eXV4TUrLC9GHuMvUWXQ62y74MARR_3U3RFCzk048oli1ieFk9czBk--rwL6ubT-ffzr7UV9efL85Or2ojMJ3qphMdXztlGDekWTuQjGKqnGsctkC4ZLiIulZSYUYd7wxjRnQdU4o2LSeWLasP-77bXTfatSl3Jhj0NvkR0p2O4PX_leB7vYk_NGmlkE3hvzvwU_y-s3nSo8-zdnuRtFCN4FjQR4FUsAZLxR8FkpZhwcpbVm8eAG_jLoWilqaYFxtwJQpI7kEmxZyTdcfTCNaz33Txm26ZJlTPfpuD0LPfCvX1v9IciQeDlfrbQx2ygcGl4hufjzBKSMNUe7_mbLGfPlkNeXww9X6Wg6hhk0qfm6_zFrgVknP2F3GU-Eg</recordid><startdate>19971201</startdate><enddate>19971201</enddate><creator>Moniruzzaman, M</creator><creator>Lai, X</creator><creator>York, S.W</creator><creator>Ingram, L.O</creator><general>American Society for Microbiology</general><scope>FBQ</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>7QL</scope><scope>7QO</scope><scope>7SN</scope><scope>7SS</scope><scope>7ST</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>SOI</scope><scope>KR7</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19971201</creationdate><title>Isolation and molecular characterization of high-performance cellobiose-fermenting spontaneous mutants of ethanologenic Escherichia coli KO11 containing the Klebsiella oxytoca casAB operon</title><author>Moniruzzaman, M ; Lai, X ; York, S.W ; Ingram, L.O</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c702t-4b7b5df9c35c14dfa832029ff4f0ea15830112d989032f5bc33c7bb39924651e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Bacteria</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biology of microorganisms of confirmed or potential industrial interest</topic><topic>Biotechnology</topic><topic>Cellobiose - metabolism</topic><topic>DECHET</topic><topic>DESECHOS</topic><topic>DNA Primers - genetics</topic><topic>DNA, Bacterial - genetics</topic><topic>ESCHERICHIA COLI</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - isolation & purification</topic><topic>Escherichia coli - metabolism</topic><topic>ETANOL</topic><topic>ETHANOL</topic><topic>Ethanol - metabolism</topic><topic>ETHANOL PRODUCTION</topic><topic>FERMENTACION</topic><topic>FERMENTATION</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genes, Bacterial</topic><topic>Genetics</topic><topic>KLEBSIELLA</topic><topic>Klebsiella - genetics</topic><topic>Microbiology</topic><topic>Mission oriented research</topic><topic>Molecular Sequence Data</topic><topic>MUTANT</topic><topic>MUTANTES</topic><topic>MUTANTS</topic><topic>Mutation</topic><topic>Operon</topic><topic>PAPEL</topic><topic>PAPIER</topic><topic>WASTES</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Moniruzzaman, M</creatorcontrib><creatorcontrib>Lai, X</creatorcontrib><creatorcontrib>York, S.W</creatorcontrib><creatorcontrib>Ingram, L.O</creatorcontrib><collection>AGRIS</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Environment Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Environment Abstracts</collection><collection>Civil Engineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Applied and Environmental Microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Moniruzzaman, M</au><au>Lai, X</au><au>York, S.W</au><au>Ingram, L.O</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation and molecular characterization of high-performance cellobiose-fermenting spontaneous mutants of ethanologenic Escherichia coli KO11 containing the Klebsiella oxytoca casAB operon</atitle><jtitle>Applied and Environmental Microbiology</jtitle><addtitle>Appl Environ Microbiol</addtitle><date>1997-12-01</date><risdate>1997</risdate><volume>63</volume><issue>12</issue><spage>4633</spage><epage>4637</epage><pages>4633-4637</pages><issn>0099-2240</issn><eissn>1098-5336</eissn><coden>AEMIDF</coden><abstract>Escherichia coli KO11 was previously constructed to produce ethanol from acid hydrolysates of hemicellulose (pentoses and hexoses) by the chromosomal integration of Zymomonas mobilis genes encoding pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB). Klebsiella oxytoca P2 was constructed in an analogous fashion for the simultaneous saccharification and fermentation of cellulose and contains PTS enzymes for cellobiose. In this study, KO11 was further engineered for the fermentation of cellulose by adding the K. oxytoca casAB genes encoding Enzyme II cellobiose and phosphobetaglucosidase. Although the two K. oxytoca genes were well expressed in cloning hosts such as DH5 alpha both were expressed poorly in E. coli KO11, a derivative of E. coli B. Spontaneous mutants which exhibited more than 15-fold-higher specific activities for cellobiose metabolism were isolated. The mutations of these mutants resided in the plasmid rather than the host. Three mutants were characterized by sequence analysis. All contained similar internal deletions which eliminated the casAB promoter and operator regions and placed the lacZ Shine-Dalgarno region immediately upstream from the casA Shine-Dalgarno region. KO11 harboring mutant plasmids (pLOI1908, pLOI1909, or pLOI1910) rapidly fermented cellobiose to ethanol, and the yield was more than 90% of the theoretical yield. Two of these strains were used with commercial cellulase to ferment mixed-waste office paper to ethanol</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>9406380</pmid><doi>10.1128/aem.63.12.4633-4637.1997</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0099-2240 |
| ispartof | Applied and Environmental Microbiology, 1997-12, Vol.63 (12), p.4633-4637 |
| issn | 0099-2240 1098-5336 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_168784 |
| source | Open Access: PubMed Central; MEDLINE; American Society for Microbiology Journals; Alma/SFX Local Collection |
| subjects | Bacteria Base Sequence Biological and medical sciences Biology of microorganisms of confirmed or potential industrial interest Biotechnology Cellobiose - metabolism DECHET DESECHOS DNA Primers - genetics DNA, Bacterial - genetics ESCHERICHIA COLI Escherichia coli - genetics Escherichia coli - isolation & purification Escherichia coli - metabolism ETANOL ETHANOL Ethanol - metabolism ETHANOL PRODUCTION FERMENTACION FERMENTATION Fundamental and applied biological sciences. Psychology Gene Expression Genes, Bacterial Genetics KLEBSIELLA Klebsiella - genetics Microbiology Mission oriented research Molecular Sequence Data MUTANT MUTANTES MUTANTS Mutation Operon PAPEL PAPIER WASTES |
| title | Isolation and molecular characterization of high-performance cellobiose-fermenting spontaneous mutants of ethanologenic Escherichia coli KO11 containing the Klebsiella oxytoca casAB operon |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-14T17%3A45%3A17IST&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=Isolation%20and%20molecular%20characterization%20of%20high-performance%20cellobiose-fermenting%20spontaneous%20mutants%20of%20ethanologenic%20Escherichia%20coli%20KO11%20containing%20the%20Klebsiella%20oxytoca%20casAB%20operon&rft.jtitle=Applied%20and%20Environmental%20Microbiology&rft.au=Moniruzzaman,%20M&rft.date=1997-12-01&rft.volume=63&rft.issue=12&rft.spage=4633&rft.epage=4637&rft.pages=4633-4637&rft.issn=0099-2240&rft.eissn=1098-5336&rft.coden=AEMIDF&rft_id=info:doi/10.1128/aem.63.12.4633-4637.1997&rft_dat=%3Cproquest_pubme%3E79475072%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=205940597&rft_id=info:pmid/9406380&rfr_iscdi=true |