Endoglucanase E, produced at high level in Escherichia coli as a lacZ′ fusion protein, is part of the Clostridium thermocellum cellulosome
The Clostridium thermocellum celE gene was expressed at high level in Escherichia coli TG1 when placed under the transcriptional and translational control of lacZ′ in pUC18; in the presence of a multicopy plasmid (pNM52) containing the lacI q gene, expression of full-length and truncated forms of ce...
Gespeichert in:
| Veröffentlicht in: | Enzyme and microbial technology 1990-09, Vol.12 (9), p.656-662 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 662 |
|---|---|
| container_issue | 9 |
| container_start_page | 656 |
| container_title | Enzyme and microbial technology |
| container_volume | 12 |
| creator | Hazlewood, Geoffrey P. Davidson, Keith Clarke, Jonathan H. Durrant, Alastair J. Hall, Judith Gilbert, Harry J. |
| description | The
Clostridium thermocellum celE gene was expressed at high level in
Escherichia coli TG1 when placed under the transcriptional and translational control of
lacZ′ in pUC18; in the presence of a multicopy plasmid (pNM52) containing the
lacI
q gene, expression of full-length and truncated forms of
celE was regulated by isopropyl-β-
d-thiogalactopyranoside. Catalytically active endoglucanase E (EGE) produced by
E. coli was subject to proteolytic processing. The main protein species produced from full-length and truncated forms of
celE was around 40 kDa in size and had an N-terminal amino acid sequence corresponding to that derived for mature EGE from the nucleotide sequence; in addition, larger species of about 75 kDa, presumably corresponding to full-size EGE, were produced by
E.coli containing the full-length
celE gene. Even after removal of the signal peptide sequence, EGE produced by
E. coli was secreted into the periplasm. Up to 157 bp could be deleted from the 5′ end of the
celE gene without affecting the catalytic activity of EGE produced by
E. coli. A polypeptide of M
, 86 kDa, immunoreactive with anti-EGE antiserum, was demonstrated in the high-molecular-weight, cellulose-binding multiprotein aggregate recoverable from
C. thermocellum culture supernatant. |
| doi_str_mv | 10.1016/0141-0229(90)90004-A |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_80241777</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>014102299090004A</els_id><sourcerecordid>15892213</sourcerecordid><originalsourceid>FETCH-LOGICAL-c424t-bf07dc4e9d72d63841b038e600714ea4bceb216d5cea12473696056e32303c023</originalsourceid><addsrcrecordid>eNqFkc-KFDEQxoMo67j6Bgo5icK2Vv5Md-eyMAzjH1jwohcvIZ1Ub0fSnTHpXvDmA_g0PpJPYnpn0ZueiuL7vqqifoQ8ZfCKAatfA5OsAs7VCwUvFQDIanePbFjbqAoUqPtk88fykDzK-UvxMCnhjJwxUdcttBvy4zC5eB0WayaTkR4u6DFFt1h01Mx08NcDDXiDgfqJHrIdMHk7eENtDJ6aTA0Nxn7-9f0n7Zfs47TGZ_TTBfWZHk2aaezpPCDdh5jn5J1fxrVPY7QYQmluSxHjiI_Jg96EjE_u6jn59Obwcf-uuvrw9v1-d1VZyeVcdT00zkpUruGuFq1kHYgWa4CGSTSys9hxVrutRcO4bEStatjWKLgAYYGLc_L8NLcc-3XBPOvR5_UOM2Fcsm6BS9Y0zX-NbNsqzpkoRnky2hRzTtjrY_KjSd80A73S0isKvaLQCvQtLb0rsWd385duRPc3dMJT9MuTjuUbNx6TztbjVOj4hHbWLvp_L_gNqvalPQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>15892213</pqid></control><display><type>article</type><title>Endoglucanase E, produced at high level in Escherichia coli as a lacZ′ fusion protein, is part of the Clostridium thermocellum cellulosome</title><source>MEDLINE</source><source>ScienceDirect Journals (5 years ago - present)</source><creator>Hazlewood, Geoffrey P. ; Davidson, Keith ; Clarke, Jonathan H. ; Durrant, Alastair J. ; Hall, Judith ; Gilbert, Harry J.</creator><creatorcontrib>Hazlewood, Geoffrey P. ; Davidson, Keith ; Clarke, Jonathan H. ; Durrant, Alastair J. ; Hall, Judith ; Gilbert, Harry J.</creatorcontrib><description>The
Clostridium thermocellum celE gene was expressed at high level in
Escherichia coli TG1 when placed under the transcriptional and translational control of
lacZ′ in pUC18; in the presence of a multicopy plasmid (pNM52) containing the
lacI
q gene, expression of full-length and truncated forms of
celE was regulated by isopropyl-β-
d-thiogalactopyranoside. Catalytically active endoglucanase E (EGE) produced by
E. coli was subject to proteolytic processing. The main protein species produced from full-length and truncated forms of
celE was around 40 kDa in size and had an N-terminal amino acid sequence corresponding to that derived for mature EGE from the nucleotide sequence; in addition, larger species of about 75 kDa, presumably corresponding to full-size EGE, were produced by
E.coli containing the full-length
celE gene. Even after removal of the signal peptide sequence, EGE produced by
E. coli was secreted into the periplasm. Up to 157 bp could be deleted from the 5′ end of the
celE gene without affecting the catalytic activity of EGE produced by
E. coli. A polypeptide of M
, 86 kDa, immunoreactive with anti-EGE antiserum, was demonstrated in the high-molecular-weight, cellulose-binding multiprotein aggregate recoverable from
C. thermocellum culture supernatant.</description><identifier>ISSN: 0141-0229</identifier><identifier>EISSN: 1879-0909</identifier><identifier>DOI: 10.1016/0141-0229(90)90004-A</identifier><identifier>PMID: 1366808</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amino Acid Sequence ; Base Sequence ; Biotechnology ; Cellulase - biosynthesis ; Cellulase - genetics ; Cellulase - secretion ; cloning ; Clostridium - enzymology ; Clostridium - genetics ; Clostridium thermocellum ; DNA Mutational Analysis ; Endoglucanase ; endoglucanase E ; Escherichia coli ; Escherichia coli - metabolism ; gene fusion ; genes ; Macromolecular Substances ; Molecular Sequence Data ; Molecular Weight ; Protein Processing, Post-Translational ; Protein Sorting Signals - physiology ; Recombinant Fusion Proteins - biosynthesis ; Recombinant Fusion Proteins - secretion ; Structure-Activity Relationship ; Subcellular Fractions - enzymology</subject><ispartof>Enzyme and microbial technology, 1990-09, Vol.12 (9), p.656-662</ispartof><rights>1990</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c424t-bf07dc4e9d72d63841b038e600714ea4bceb216d5cea12473696056e32303c023</citedby><cites>FETCH-LOGICAL-c424t-bf07dc4e9d72d63841b038e600714ea4bceb216d5cea12473696056e32303c023</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0141-0229(90)90004-A$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3549,27923,27924,45994</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1366808$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hazlewood, Geoffrey P.</creatorcontrib><creatorcontrib>Davidson, Keith</creatorcontrib><creatorcontrib>Clarke, Jonathan H.</creatorcontrib><creatorcontrib>Durrant, Alastair J.</creatorcontrib><creatorcontrib>Hall, Judith</creatorcontrib><creatorcontrib>Gilbert, Harry J.</creatorcontrib><title>Endoglucanase E, produced at high level in Escherichia coli as a lacZ′ fusion protein, is part of the Clostridium thermocellum cellulosome</title><title>Enzyme and microbial technology</title><addtitle>Enzyme Microb Technol</addtitle><description>The
Clostridium thermocellum celE gene was expressed at high level in
Escherichia coli TG1 when placed under the transcriptional and translational control of
lacZ′ in pUC18; in the presence of a multicopy plasmid (pNM52) containing the
lacI
q gene, expression of full-length and truncated forms of
celE was regulated by isopropyl-β-
d-thiogalactopyranoside. Catalytically active endoglucanase E (EGE) produced by
E. coli was subject to proteolytic processing. The main protein species produced from full-length and truncated forms of
celE was around 40 kDa in size and had an N-terminal amino acid sequence corresponding to that derived for mature EGE from the nucleotide sequence; in addition, larger species of about 75 kDa, presumably corresponding to full-size EGE, were produced by
E.coli containing the full-length
celE gene. Even after removal of the signal peptide sequence, EGE produced by
E. coli was secreted into the periplasm. Up to 157 bp could be deleted from the 5′ end of the
celE gene without affecting the catalytic activity of EGE produced by
E. coli. A polypeptide of M
, 86 kDa, immunoreactive with anti-EGE antiserum, was demonstrated in the high-molecular-weight, cellulose-binding multiprotein aggregate recoverable from
C. thermocellum culture supernatant.</description><subject>Amino Acid Sequence</subject><subject>Base Sequence</subject><subject>Biotechnology</subject><subject>Cellulase - biosynthesis</subject><subject>Cellulase - genetics</subject><subject>Cellulase - secretion</subject><subject>cloning</subject><subject>Clostridium - enzymology</subject><subject>Clostridium - genetics</subject><subject>Clostridium thermocellum</subject><subject>DNA Mutational Analysis</subject><subject>Endoglucanase</subject><subject>endoglucanase E</subject><subject>Escherichia coli</subject><subject>Escherichia coli - metabolism</subject><subject>gene fusion</subject><subject>genes</subject><subject>Macromolecular Substances</subject><subject>Molecular Sequence Data</subject><subject>Molecular Weight</subject><subject>Protein Processing, Post-Translational</subject><subject>Protein Sorting Signals - physiology</subject><subject>Recombinant Fusion Proteins - biosynthesis</subject><subject>Recombinant Fusion Proteins - secretion</subject><subject>Structure-Activity Relationship</subject><subject>Subcellular Fractions - enzymology</subject><issn>0141-0229</issn><issn>1879-0909</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc-KFDEQxoMo67j6Bgo5icK2Vv5Md-eyMAzjH1jwohcvIZ1Ub0fSnTHpXvDmA_g0PpJPYnpn0ZueiuL7vqqifoQ8ZfCKAatfA5OsAs7VCwUvFQDIanePbFjbqAoUqPtk88fykDzK-UvxMCnhjJwxUdcttBvy4zC5eB0WayaTkR4u6DFFt1h01Mx08NcDDXiDgfqJHrIdMHk7eENtDJ6aTA0Nxn7-9f0n7Zfs47TGZ_TTBfWZHk2aaezpPCDdh5jn5J1fxrVPY7QYQmluSxHjiI_Jg96EjE_u6jn59Obwcf-uuvrw9v1-d1VZyeVcdT00zkpUruGuFq1kHYgWa4CGSTSys9hxVrutRcO4bEStatjWKLgAYYGLc_L8NLcc-3XBPOvR5_UOM2Fcsm6BS9Y0zX-NbNsqzpkoRnky2hRzTtjrY_KjSd80A73S0isKvaLQCvQtLb0rsWd385duRPc3dMJT9MuTjuUbNx6TztbjVOj4hHbWLvp_L_gNqvalPQ</recordid><startdate>19900901</startdate><enddate>19900901</enddate><creator>Hazlewood, Geoffrey P.</creator><creator>Davidson, Keith</creator><creator>Clarke, Jonathan H.</creator><creator>Durrant, Alastair J.</creator><creator>Hall, Judith</creator><creator>Gilbert, Harry J.</creator><general>Elsevier Inc</general><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>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19900901</creationdate><title>Endoglucanase E, produced at high level in Escherichia coli as a lacZ′ fusion protein, is part of the Clostridium thermocellum cellulosome</title><author>Hazlewood, Geoffrey P. ; Davidson, Keith ; Clarke, Jonathan H. ; Durrant, Alastair J. ; Hall, Judith ; Gilbert, Harry J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c424t-bf07dc4e9d72d63841b038e600714ea4bceb216d5cea12473696056e32303c023</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Amino Acid Sequence</topic><topic>Base Sequence</topic><topic>Biotechnology</topic><topic>Cellulase - biosynthesis</topic><topic>Cellulase - genetics</topic><topic>Cellulase - secretion</topic><topic>cloning</topic><topic>Clostridium - enzymology</topic><topic>Clostridium - genetics</topic><topic>Clostridium thermocellum</topic><topic>DNA Mutational Analysis</topic><topic>Endoglucanase</topic><topic>endoglucanase E</topic><topic>Escherichia coli</topic><topic>Escherichia coli - metabolism</topic><topic>gene fusion</topic><topic>genes</topic><topic>Macromolecular Substances</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>Protein Processing, Post-Translational</topic><topic>Protein Sorting Signals - physiology</topic><topic>Recombinant Fusion Proteins - biosynthesis</topic><topic>Recombinant Fusion Proteins - secretion</topic><topic>Structure-Activity Relationship</topic><topic>Subcellular Fractions - enzymology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hazlewood, Geoffrey P.</creatorcontrib><creatorcontrib>Davidson, Keith</creatorcontrib><creatorcontrib>Clarke, Jonathan H.</creatorcontrib><creatorcontrib>Durrant, Alastair J.</creatorcontrib><creatorcontrib>Hall, Judith</creatorcontrib><creatorcontrib>Gilbert, Harry J.</creatorcontrib><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>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</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>Hazlewood, Geoffrey P.</au><au>Davidson, Keith</au><au>Clarke, Jonathan H.</au><au>Durrant, Alastair J.</au><au>Hall, Judith</au><au>Gilbert, Harry J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Endoglucanase E, produced at high level in Escherichia coli as a lacZ′ fusion protein, is part of the Clostridium thermocellum cellulosome</atitle><jtitle>Enzyme and microbial technology</jtitle><addtitle>Enzyme Microb Technol</addtitle><date>1990-09-01</date><risdate>1990</risdate><volume>12</volume><issue>9</issue><spage>656</spage><epage>662</epage><pages>656-662</pages><issn>0141-0229</issn><eissn>1879-0909</eissn><abstract>The
Clostridium thermocellum celE gene was expressed at high level in
Escherichia coli TG1 when placed under the transcriptional and translational control of
lacZ′ in pUC18; in the presence of a multicopy plasmid (pNM52) containing the
lacI
q gene, expression of full-length and truncated forms of
celE was regulated by isopropyl-β-
d-thiogalactopyranoside. Catalytically active endoglucanase E (EGE) produced by
E. coli was subject to proteolytic processing. The main protein species produced from full-length and truncated forms of
celE was around 40 kDa in size and had an N-terminal amino acid sequence corresponding to that derived for mature EGE from the nucleotide sequence; in addition, larger species of about 75 kDa, presumably corresponding to full-size EGE, were produced by
E.coli containing the full-length
celE gene. Even after removal of the signal peptide sequence, EGE produced by
E. coli was secreted into the periplasm. Up to 157 bp could be deleted from the 5′ end of the
celE gene without affecting the catalytic activity of EGE produced by
E. coli. A polypeptide of M
, 86 kDa, immunoreactive with anti-EGE antiserum, was demonstrated in the high-molecular-weight, cellulose-binding multiprotein aggregate recoverable from
C. thermocellum culture supernatant.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>1366808</pmid><doi>10.1016/0141-0229(90)90004-A</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0141-0229 |
| ispartof | Enzyme and microbial technology, 1990-09, Vol.12 (9), p.656-662 |
| issn | 0141-0229 1879-0909 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_80241777 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | Amino Acid Sequence Base Sequence Biotechnology Cellulase - biosynthesis Cellulase - genetics Cellulase - secretion cloning Clostridium - enzymology Clostridium - genetics Clostridium thermocellum DNA Mutational Analysis Endoglucanase endoglucanase E Escherichia coli Escherichia coli - metabolism gene fusion genes Macromolecular Substances Molecular Sequence Data Molecular Weight Protein Processing, Post-Translational Protein Sorting Signals - physiology Recombinant Fusion Proteins - biosynthesis Recombinant Fusion Proteins - secretion Structure-Activity Relationship Subcellular Fractions - enzymology |
| title | Endoglucanase E, produced at high level in Escherichia coli as a lacZ′ fusion protein, is part of the Clostridium thermocellum cellulosome |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-08T18%3A42%3A26IST&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=Endoglucanase%20E,%20produced%20at%20high%20level%20in%20Escherichia%20coli%20as%20a%20lacZ%E2%80%B2%20fusion%20protein,%20is%20part%20of%20the%20Clostridium%20thermocellum%20cellulosome&rft.jtitle=Enzyme%20and%20microbial%20technology&rft.au=Hazlewood,%20Geoffrey%20P.&rft.date=1990-09-01&rft.volume=12&rft.issue=9&rft.spage=656&rft.epage=662&rft.pages=656-662&rft.issn=0141-0229&rft.eissn=1879-0909&rft_id=info:doi/10.1016/0141-0229(90)90004-A&rft_dat=%3Cproquest_cross%3E15892213%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=15892213&rft_id=info:pmid/1366808&rft_els_id=014102299090004A&rfr_iscdi=true |