Cloning of a Gene Encoding β-Glucosidase from Chaetomium thermophilum CT2 and Its Expression in Pichia pastoris
A new thermostable β-glucosidase gene (bgl) from Chaetomium thermophilum CT2 was cloned, sequenced and expressed. The full-length DNA of bgl was 3,101 bp and included three introns. The full-length cDNA contained an open reading frame of 2,604-bp nucleotides, encoding 867 amino acids with a potentia...
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| Veröffentlicht in: | Journal of molecular microbiology and biotechnology 2011-01, Vol.20 (1), p.16-23 |
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| creator | Xu, Rongyan Teng, Fangchao Zhang, Chao Li, Duochuan |
| description | A new thermostable β-glucosidase gene (bgl) from Chaetomium thermophilum CT2 was cloned, sequenced and expressed. The full-length DNA of bgl was 3,101 bp and included three introns. The full-length cDNA contained an open reading frame of 2,604-bp nucleotides, encoding 867 amino acids with a potential secretion signal. The C. thermophilum CT2 β-glucosidase gene was functionally expressed in Pichia pastoris. The purified recombinant β-glucosidase was a 119-kDa glycoprotein with an optimum catalytic activity at pH 5.0 and 60°C. The enzyme was stable at 50°C, and retained 67.7% activity after being kept at 60°C for 1 h; the half-time of the enzyme at 65°C was approximately 55 min, and even retained 29.7% activity after incubation at 70°C for 10 min. |
| doi_str_mv | 10.1159/000322606 |
| format | Article |
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The full-length DNA of bgl was 3,101 bp and included three introns. The full-length cDNA contained an open reading frame of 2,604-bp nucleotides, encoding 867 amino acids with a potential secretion signal. The C. thermophilum CT2 β-glucosidase gene was functionally expressed in Pichia pastoris. The purified recombinant β-glucosidase was a 119-kDa glycoprotein with an optimum catalytic activity at pH 5.0 and 60°C. The enzyme was stable at 50°C, and retained 67.7% activity after being kept at 60°C for 1 h; the half-time of the enzyme at 65°C was approximately 55 min, and even retained 29.7% activity after incubation at 70°C for 10 min.</description><identifier>ISSN: 2673-1665</identifier><identifier>EISSN: 2673-1673</identifier><identifier>EISSN: 1660-2412</identifier><identifier>DOI: 10.1159/000322606</identifier><identifier>PMID: 21273791</identifier><language>eng</language><publisher>Basel, Switzerland</publisher><subject>Amino Acid Sequence ; beta-Glucosidase - chemistry ; beta-Glucosidase - genetics ; beta-Glucosidase - metabolism ; Chaetomium - enzymology ; Chaetomium - genetics ; Cloning, Molecular ; DNA, Fungal - chemistry ; DNA, Fungal - genetics ; Enzyme Stability ; Gene Expression ; Hydrogen-Ion Concentration ; Introns ; Molecular Sequence Data ; Molecular Weight ; Open Reading Frames ; Pichia - genetics ; Protein Sorting Signals ; Recombinant Proteins - chemistry ; Recombinant Proteins - genetics ; Recombinant Proteins - isolation & purification ; Recombinant Proteins - metabolism ; Research Article ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Temperature</subject><ispartof>Journal of molecular microbiology and biotechnology, 2011-01, Vol.20 (1), p.16-23</ispartof><rights>2011 S. Karger AG, Basel</rights><rights>Copyright © 2011 S. Karger AG, Basel.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c305t-868a1de931c5fa6297162c697fbcfd020279775a4e97f071d9f817a5d152627c3</citedby><cites>FETCH-LOGICAL-c305t-868a1de931c5fa6297162c697fbcfd020279775a4e97f071d9f817a5d152627c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,2423,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21273791$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Xu, Rongyan</creatorcontrib><creatorcontrib>Teng, Fangchao</creatorcontrib><creatorcontrib>Zhang, Chao</creatorcontrib><creatorcontrib>Li, Duochuan</creatorcontrib><title>Cloning of a Gene Encoding β-Glucosidase from Chaetomium thermophilum CT2 and Its Expression in Pichia pastoris</title><title>Journal of molecular microbiology and biotechnology</title><addtitle>Microb Physiol</addtitle><description>A new thermostable β-glucosidase gene (bgl) from Chaetomium thermophilum CT2 was cloned, sequenced and expressed. The full-length DNA of bgl was 3,101 bp and included three introns. The full-length cDNA contained an open reading frame of 2,604-bp nucleotides, encoding 867 amino acids with a potential secretion signal. The C. thermophilum CT2 β-glucosidase gene was functionally expressed in Pichia pastoris. The purified recombinant β-glucosidase was a 119-kDa glycoprotein with an optimum catalytic activity at pH 5.0 and 60°C. 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Teng, Fangchao ; Zhang, Chao ; Li, Duochuan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c305t-868a1de931c5fa6297162c697fbcfd020279775a4e97f071d9f817a5d152627c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Amino Acid Sequence</topic><topic>beta-Glucosidase - chemistry</topic><topic>beta-Glucosidase - genetics</topic><topic>beta-Glucosidase - metabolism</topic><topic>Chaetomium - enzymology</topic><topic>Chaetomium - genetics</topic><topic>Cloning, Molecular</topic><topic>DNA, Fungal - chemistry</topic><topic>DNA, Fungal - genetics</topic><topic>Enzyme Stability</topic><topic>Gene Expression</topic><topic>Hydrogen-Ion Concentration</topic><topic>Introns</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>Open Reading Frames</topic><topic>Pichia - genetics</topic><topic>Protein Sorting Signals</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Recombinant Proteins - metabolism</topic><topic>Research Article</topic><topic>Sequence Analysis, DNA</topic><topic>Sequence Homology, Amino Acid</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Xu, Rongyan</creatorcontrib><creatorcontrib>Teng, Fangchao</creatorcontrib><creatorcontrib>Zhang, Chao</creatorcontrib><creatorcontrib>Li, Duochuan</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of molecular microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xu, Rongyan</au><au>Teng, Fangchao</au><au>Zhang, Chao</au><au>Li, Duochuan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning of a Gene Encoding β-Glucosidase from Chaetomium thermophilum CT2 and Its Expression in Pichia pastoris</atitle><jtitle>Journal of molecular microbiology and biotechnology</jtitle><addtitle>Microb Physiol</addtitle><date>2011-01-01</date><risdate>2011</risdate><volume>20</volume><issue>1</issue><spage>16</spage><epage>23</epage><pages>16-23</pages><issn>2673-1665</issn><eissn>2673-1673</eissn><eissn>1660-2412</eissn><abstract>A new thermostable β-glucosidase gene (bgl) from Chaetomium thermophilum CT2 was cloned, sequenced and expressed. The full-length DNA of bgl was 3,101 bp and included three introns. The full-length cDNA contained an open reading frame of 2,604-bp nucleotides, encoding 867 amino acids with a potential secretion signal. The C. thermophilum CT2 β-glucosidase gene was functionally expressed in Pichia pastoris. The purified recombinant β-glucosidase was a 119-kDa glycoprotein with an optimum catalytic activity at pH 5.0 and 60°C. The enzyme was stable at 50°C, and retained 67.7% activity after being kept at 60°C for 1 h; the half-time of the enzyme at 65°C was approximately 55 min, and even retained 29.7% activity after incubation at 70°C for 10 min.</abstract><cop>Basel, Switzerland</cop><pmid>21273791</pmid><doi>10.1159/000322606</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Journal of molecular microbiology and biotechnology, 2011-01, Vol.20 (1), p.16-23 |
| issn | 2673-1665 2673-1673 1660-2412 |
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| source | Karger Journals; MEDLINE; Alma/SFX Local Collection |
| subjects | Amino Acid Sequence beta-Glucosidase - chemistry beta-Glucosidase - genetics beta-Glucosidase - metabolism Chaetomium - enzymology Chaetomium - genetics Cloning, Molecular DNA, Fungal - chemistry DNA, Fungal - genetics Enzyme Stability Gene Expression Hydrogen-Ion Concentration Introns Molecular Sequence Data Molecular Weight Open Reading Frames Pichia - genetics Protein Sorting Signals Recombinant Proteins - chemistry Recombinant Proteins - genetics Recombinant Proteins - isolation & purification Recombinant Proteins - metabolism Research Article Sequence Analysis, DNA Sequence Homology, Amino Acid Temperature |
| title | Cloning of a Gene Encoding β-Glucosidase from Chaetomium thermophilum CT2 and Its Expression in Pichia pastoris |
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