Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda
► Cel8A from Klebsiella pneumoniae XM-4 has firstly been cloned and expressed in Escherichia coli. ► The highest recombinant protein was obtained by used pET-32a(+) without signal peptide to cel8A. ► 95% similarity of Cel8A to Cellulomonas uda showed the biological evolution between both strains. Kl...
Gespeichert in:
| Veröffentlicht in: | Biochemical engineering journal 2013-09, Vol.78, p.53-58 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 58 |
|---|---|
| container_issue | |
| container_start_page | 53 |
| container_title | Biochemical engineering journal |
| container_volume | 78 |
| creator | Ng, I-Son Chi, Xiaoqin Wu, Xiaomin Bao, Ziwei Lu, Yinghua Chang, Jo-Shu Ling, Xueping |
| description | ► Cel8A from Klebsiella pneumoniae XM-4 has firstly been cloned and expressed in Escherichia coli. ► The highest recombinant protein was obtained by used pET-32a(+) without signal peptide to cel8A. ► 95% similarity of Cel8A to Cellulomonas uda showed the biological evolution between both strains.
Klebsiella pneumoniae XM-4 is first isolated from a bacterial consortium. According to the 16S rRNA sequence, it has only 75% sequence similarity to Cellulomonas sp. However, a 999-bp open reading frame of K. pneumoniae, which encodes a putative endo-glucanase of 333 amino acids belonging to GH8 family and designated as Cel8A, is found closely to Cellulomonas uda with 95% similarity in amino acid sequence. This implies that the biological evolution between both strains has occurred. The cel8A is constructed in vectors of pET-22b(+) and pET-32a(+) with or without containing a signal peptide, and then cloned and expressed in Escherichia coli. The best endo-glucanase production of 25.4mg/l is then obtainable by applying pET-32a(+) without the signal peptide. The recombinant CelA8 has an optimal specific activity of 62.4U/mg against CMC or 23.3U/mg against β-d-glucan at 55°C and pH 5.0, indicating good potential for the industrial application. |
| doi_str_mv | 10.1016/j.bej.2013.01.009 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1500794801</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S1369703X1300020X</els_id><sourcerecordid>1500794801</sourcerecordid><originalsourceid>FETCH-LOGICAL-c384t-f1f65920e08c55066c9509081954c133537cdc81803a841b24c06261087a35f33</originalsourceid><addsrcrecordid>eNp9kT2P1DAQhi0EEsfCD6DCDRJNwjiOPyKq0-r4ECdRwEnXWV5nsueVYwd7g6Dgv-NlV5RUnuJ9H42fIeQlg5YBk28P7Q4PbQeMt8BagOERuWJa8aYbxP3jOnM5NAr4_VPyrJQDAEiu1BX5vQ0p-rinNo4Ufy4ZS_Ep0jTRLQZ9TaecZvo54K54DMHSJeI614pF6iO9Ke4Bs3cP3lKXgv-LcWlebPalYo6JOgx0jxEvyLCGVPu20HW0z8mTyYaCLy7vhty9v_m2_djcfvnwaXt92ziu-2MzsUmKoQME7YQAKd0gYADNBtE7xrngyo1OMw3c6p7tut6B7CQDrSwXE-cb8ubMXXL6vmI5mtkXd_pPxLQWwwSAGnpd9W0IO0ddTqVknMyS_WzzL8PAnFSbg6mqzUm1AWaq6tp5fcHb4myYso3Ol3_FTqlO80HV3KtzbrLJ2H1VZO6-VpCs5xA9Y7Im3p0TWG388JhNcR6jw9FndEczJv-fPf4AaMGb0w</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1500794801</pqid></control><display><type>article</type><title>Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda</title><source>Elsevier ScienceDirect Journals</source><creator>Ng, I-Son ; Chi, Xiaoqin ; Wu, Xiaomin ; Bao, Ziwei ; Lu, Yinghua ; Chang, Jo-Shu ; Ling, Xueping</creator><creatorcontrib>Ng, I-Son ; Chi, Xiaoqin ; Wu, Xiaomin ; Bao, Ziwei ; Lu, Yinghua ; Chang, Jo-Shu ; Ling, Xueping</creatorcontrib><description>► Cel8A from Klebsiella pneumoniae XM-4 has firstly been cloned and expressed in Escherichia coli. ► The highest recombinant protein was obtained by used pET-32a(+) without signal peptide to cel8A. ► 95% similarity of Cel8A to Cellulomonas uda showed the biological evolution between both strains.
Klebsiella pneumoniae XM-4 is first isolated from a bacterial consortium. According to the 16S rRNA sequence, it has only 75% sequence similarity to Cellulomonas sp. However, a 999-bp open reading frame of K. pneumoniae, which encodes a putative endo-glucanase of 333 amino acids belonging to GH8 family and designated as Cel8A, is found closely to Cellulomonas uda with 95% similarity in amino acid sequence. This implies that the biological evolution between both strains has occurred. The cel8A is constructed in vectors of pET-22b(+) and pET-32a(+) with or without containing a signal peptide, and then cloned and expressed in Escherichia coli. The best endo-glucanase production of 25.4mg/l is then obtainable by applying pET-32a(+) without the signal peptide. The recombinant CelA8 has an optimal specific activity of 62.4U/mg against CMC or 23.3U/mg against β-d-glucan at 55°C and pH 5.0, indicating good potential for the industrial application.</description><identifier>ISSN: 1369-703X</identifier><identifier>EISSN: 1873-295X</identifier><identifier>DOI: 10.1016/j.bej.2013.01.009</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>amino acids ; beta-glucans ; Biological and medical sciences ; Biological evolution ; Biotechnology ; Cellulomonas ; Cellulomonas uda ; Endo-glucanase ; Escherichia coli ; evolution ; Fundamental and applied biological sciences. Psychology ; genes ; industrial applications ; Klebsiella pneumoniae ; open reading frames ; Recombinant expression ; ribosomal RNA ; sequence homology ; Signal peptide</subject><ispartof>Biochemical engineering journal, 2013-09, Vol.78, p.53-58</ispartof><rights>2013</rights><rights>2014 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c384t-f1f65920e08c55066c9509081954c133537cdc81803a841b24c06261087a35f33</citedby><cites>FETCH-LOGICAL-c384t-f1f65920e08c55066c9509081954c133537cdc81803a841b24c06261087a35f33</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S1369703X1300020X$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>309,310,314,776,780,785,786,3537,23909,23910,25118,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=27728397$$DView record in Pascal Francis$$Hfree_for_read</backlink></links><search><creatorcontrib>Ng, I-Son</creatorcontrib><creatorcontrib>Chi, Xiaoqin</creatorcontrib><creatorcontrib>Wu, Xiaomin</creatorcontrib><creatorcontrib>Bao, Ziwei</creatorcontrib><creatorcontrib>Lu, Yinghua</creatorcontrib><creatorcontrib>Chang, Jo-Shu</creatorcontrib><creatorcontrib>Ling, Xueping</creatorcontrib><title>Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda</title><title>Biochemical engineering journal</title><description>► Cel8A from Klebsiella pneumoniae XM-4 has firstly been cloned and expressed in Escherichia coli. ► The highest recombinant protein was obtained by used pET-32a(+) without signal peptide to cel8A. ► 95% similarity of Cel8A to Cellulomonas uda showed the biological evolution between both strains.
Klebsiella pneumoniae XM-4 is first isolated from a bacterial consortium. According to the 16S rRNA sequence, it has only 75% sequence similarity to Cellulomonas sp. However, a 999-bp open reading frame of K. pneumoniae, which encodes a putative endo-glucanase of 333 amino acids belonging to GH8 family and designated as Cel8A, is found closely to Cellulomonas uda with 95% similarity in amino acid sequence. This implies that the biological evolution between both strains has occurred. The cel8A is constructed in vectors of pET-22b(+) and pET-32a(+) with or without containing a signal peptide, and then cloned and expressed in Escherichia coli. The best endo-glucanase production of 25.4mg/l is then obtainable by applying pET-32a(+) without the signal peptide. The recombinant CelA8 has an optimal specific activity of 62.4U/mg against CMC or 23.3U/mg against β-d-glucan at 55°C and pH 5.0, indicating good potential for the industrial application.</description><subject>amino acids</subject><subject>beta-glucans</subject><subject>Biological and medical sciences</subject><subject>Biological evolution</subject><subject>Biotechnology</subject><subject>Cellulomonas</subject><subject>Cellulomonas uda</subject><subject>Endo-glucanase</subject><subject>Escherichia coli</subject><subject>evolution</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>genes</subject><subject>industrial applications</subject><subject>Klebsiella pneumoniae</subject><subject>open reading frames</subject><subject>Recombinant expression</subject><subject>ribosomal RNA</subject><subject>sequence homology</subject><subject>Signal peptide</subject><issn>1369-703X</issn><issn>1873-295X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><recordid>eNp9kT2P1DAQhi0EEsfCD6DCDRJNwjiOPyKq0-r4ECdRwEnXWV5nsueVYwd7g6Dgv-NlV5RUnuJ9H42fIeQlg5YBk28P7Q4PbQeMt8BagOERuWJa8aYbxP3jOnM5NAr4_VPyrJQDAEiu1BX5vQ0p-rinNo4Ufy4ZS_Ep0jTRLQZ9TaecZvo54K54DMHSJeI614pF6iO9Ke4Bs3cP3lKXgv-LcWlebPalYo6JOgx0jxEvyLCGVPu20HW0z8mTyYaCLy7vhty9v_m2_djcfvnwaXt92ziu-2MzsUmKoQME7YQAKd0gYADNBtE7xrngyo1OMw3c6p7tut6B7CQDrSwXE-cb8ubMXXL6vmI5mtkXd_pPxLQWwwSAGnpd9W0IO0ddTqVknMyS_WzzL8PAnFSbg6mqzUm1AWaq6tp5fcHb4myYso3Ol3_FTqlO80HV3KtzbrLJ2H1VZO6-VpCs5xA9Y7Im3p0TWG388JhNcR6jw9FndEczJv-fPf4AaMGb0w</recordid><startdate>20130915</startdate><enddate>20130915</enddate><creator>Ng, I-Son</creator><creator>Chi, Xiaoqin</creator><creator>Wu, Xiaomin</creator><creator>Bao, Ziwei</creator><creator>Lu, Yinghua</creator><creator>Chang, Jo-Shu</creator><creator>Ling, Xueping</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>FBQ</scope><scope>IQODW</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>RC3</scope></search><sort><creationdate>20130915</creationdate><title>Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda</title><author>Ng, I-Son ; Chi, Xiaoqin ; Wu, Xiaomin ; Bao, Ziwei ; Lu, Yinghua ; Chang, Jo-Shu ; Ling, Xueping</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c384t-f1f65920e08c55066c9509081954c133537cdc81803a841b24c06261087a35f33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>amino acids</topic><topic>beta-glucans</topic><topic>Biological and medical sciences</topic><topic>Biological evolution</topic><topic>Biotechnology</topic><topic>Cellulomonas</topic><topic>Cellulomonas uda</topic><topic>Endo-glucanase</topic><topic>Escherichia coli</topic><topic>evolution</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>genes</topic><topic>industrial applications</topic><topic>Klebsiella pneumoniae</topic><topic>open reading frames</topic><topic>Recombinant expression</topic><topic>ribosomal RNA</topic><topic>sequence homology</topic><topic>Signal peptide</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ng, I-Son</creatorcontrib><creatorcontrib>Chi, Xiaoqin</creatorcontrib><creatorcontrib>Wu, Xiaomin</creatorcontrib><creatorcontrib>Bao, Ziwei</creatorcontrib><creatorcontrib>Lu, Yinghua</creatorcontrib><creatorcontrib>Chang, Jo-Shu</creatorcontrib><creatorcontrib>Ling, Xueping</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</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>Genetics Abstracts</collection><jtitle>Biochemical engineering journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ng, I-Son</au><au>Chi, Xiaoqin</au><au>Wu, Xiaomin</au><au>Bao, Ziwei</au><au>Lu, Yinghua</au><au>Chang, Jo-Shu</au><au>Ling, Xueping</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda</atitle><jtitle>Biochemical engineering journal</jtitle><date>2013-09-15</date><risdate>2013</risdate><volume>78</volume><spage>53</spage><epage>58</epage><pages>53-58</pages><issn>1369-703X</issn><eissn>1873-295X</eissn><abstract>► Cel8A from Klebsiella pneumoniae XM-4 has firstly been cloned and expressed in Escherichia coli. ► The highest recombinant protein was obtained by used pET-32a(+) without signal peptide to cel8A. ► 95% similarity of Cel8A to Cellulomonas uda showed the biological evolution between both strains.
Klebsiella pneumoniae XM-4 is first isolated from a bacterial consortium. According to the 16S rRNA sequence, it has only 75% sequence similarity to Cellulomonas sp. However, a 999-bp open reading frame of K. pneumoniae, which encodes a putative endo-glucanase of 333 amino acids belonging to GH8 family and designated as Cel8A, is found closely to Cellulomonas uda with 95% similarity in amino acid sequence. This implies that the biological evolution between both strains has occurred. The cel8A is constructed in vectors of pET-22b(+) and pET-32a(+) with or without containing a signal peptide, and then cloned and expressed in Escherichia coli. The best endo-glucanase production of 25.4mg/l is then obtainable by applying pET-32a(+) without the signal peptide. The recombinant CelA8 has an optimal specific activity of 62.4U/mg against CMC or 23.3U/mg against β-d-glucan at 55°C and pH 5.0, indicating good potential for the industrial application.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><doi>10.1016/j.bej.2013.01.009</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1369-703X |
| ispartof | Biochemical engineering journal, 2013-09, Vol.78, p.53-58 |
| issn | 1369-703X 1873-295X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1500794801 |
| source | Elsevier ScienceDirect Journals |
| subjects | amino acids beta-glucans Biological and medical sciences Biological evolution Biotechnology Cellulomonas Cellulomonas uda Endo-glucanase Escherichia coli evolution Fundamental and applied biological sciences. Psychology genes industrial applications Klebsiella pneumoniae open reading frames Recombinant expression ribosomal RNA sequence homology Signal peptide |
| title | Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-05T05%3A29%3A21IST&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=Cloning%20and%20expression%20of%20Cel8A%20from%20Klebsiella%20pneumoniae%20in%20Escherichia%20coli%20and%20comparison%20to%20cel%20gene%20of%20Cellulomonas%20uda&rft.jtitle=Biochemical%20engineering%20journal&rft.au=Ng,%20I-Son&rft.date=2013-09-15&rft.volume=78&rft.spage=53&rft.epage=58&rft.pages=53-58&rft.issn=1369-703X&rft.eissn=1873-295X&rft_id=info:doi/10.1016/j.bej.2013.01.009&rft_dat=%3Cproquest_cross%3E1500794801%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=1500794801&rft_id=info:pmid/&rft_els_id=S1369703X1300020X&rfr_iscdi=true |