Characteristics of bifunctional acidic endoglucanase (Cel5B) from Gloeophyllum trabeum
The endoglucanase (Cel5B) from the filamentous fungus Gloeophyllum trabeum was cloned and expressed without a signal peptide, and alanine residue 22 converted to glutamine in Pichia pastoris GS115. The DNA sequence of Cel5B had an open reading frame of 1,077 bp, encoding a protein of 359 amino acid...
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| creator | Kim, Ho Myeong Lee, Yoon Gyo Patel, Darshan H Lee, Kwang Ho Lee, Dae-Seok Bae, Hyeun-Jong |
| description | The endoglucanase (Cel5B) from the filamentous fungus Gloeophyllum trabeum was cloned and expressed without a signal peptide, and alanine residue 22 converted to glutamine in Pichia pastoris GS115. The DNA sequence of Cel5B had an open reading frame of 1,077 bp, encoding a protein of 359 amino acid residues with a molecular weight of 47 kDa. On the basis of sequence similarity, Cel5B displayed active site residues at Glu-175 and Glu-287. Both residues lost full hydrolytic activity when replaced with alanine through point mutation. The purified recombinant Cel5B showed very high specific activity, about 80- to 1,000-fold and 13- to 70-fold in comparison with other endoglucanases and cellobiohydrolase, on carboxymethylcellulose and filter paper, respectively, at pH 3.5 and 55°C. Cel5B displayed bifunctional characteristics under acidic conditions. The kinetic properties of the enzyme determined using a Lineweaver–Burk plot indicated that Cel5B is a catalytically efficient cellulolytic enzyme. These results suggest that Cel5B has high bifunctional endo- and exoglucanase activity under acidic conditions and is a good candidate for bioconversion of lignocellulose. |
| doi_str_mv | 10.1007/s10295-012-1110-4 |
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The DNA sequence of Cel5B had an open reading frame of 1,077 bp, encoding a protein of 359 amino acid residues with a molecular weight of 47 kDa. On the basis of sequence similarity, Cel5B displayed active site residues at Glu-175 and Glu-287. Both residues lost full hydrolytic activity when replaced with alanine through point mutation. The purified recombinant Cel5B showed very high specific activity, about 80- to 1,000-fold and 13- to 70-fold in comparison with other endoglucanases and cellobiohydrolase, on carboxymethylcellulose and filter paper, respectively, at pH 3.5 and 55°C. Cel5B displayed bifunctional characteristics under acidic conditions. The kinetic properties of the enzyme determined using a Lineweaver–Burk plot indicated that Cel5B is a catalytically efficient cellulolytic enzyme. These results suggest that Cel5B has high bifunctional endo- and exoglucanase activity under acidic conditions and is a good candidate for bioconversion of lignocellulose.</description><identifier>ISSN: 1367-5435</identifier><identifier>EISSN: 1476-5535</identifier><identifier>DOI: 10.1007/s10295-012-1110-4</identifier><identifier>PMID: 22395898</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer-Verlag</publisher><subject>alanine ; Alternative energy sources ; Amino Acid Sequence ; Amino acids ; Basidiomycota - enzymology ; Basidiomycota - genetics ; Biochemistry ; Bioinformatics ; Biological and medical sciences ; Biomass ; Biomedical and Life Sciences ; Biotechnology ; biotransformation ; carboxymethylcellulose ; Catalytic Domain ; Cellulase ; Cellulase - chemistry ; Cellulase - genetics ; Cellulase - isolation & purification ; Cellulase - metabolism ; Cellulose ; cellulose 1,4-beta-cellobiosidase ; Cellulose 1,4-beta-Cellobiosidase - genetics ; Cellulose 1,4-beta-Cellobiosidase - metabolism ; Cloning ; Cloning, Molecular ; DNA polymerase ; endo-1,4-beta-glucanase ; Enzymes ; Escherichia coli - genetics ; Fossil fuels ; Fundamental and applied biological sciences. Psychology ; fungi ; Genetic Engineering ; Genetics and Molecular Biology of Industrial Organisms ; Gloeophyllum trabeum ; glutamine ; Industrial Microbiology ; Inorganic Chemistry ; Life Sciences ; lignocellulose ; Microbiology ; Microorganisms ; Models, Molecular ; Molecular Sequence Data ; molecular weight ; Mutation ; open reading frames ; Peptides ; Pichia - genetics ; Pichia pastoris ; point mutation ; Protein expression ; Proteins ; Residues ; Sequence Alignment ; signal peptide ; Yeast</subject><ispartof>Journal of industrial microbiology & biotechnology, 2012-07, Vol.39 (7), p.1081-1089</ispartof><rights>Society for Industrial Microbiology and Biotechnology 2012</rights><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c502t-494cb8a26d1955845295ef2ee2268a428bb420b22b53f5a6892dde9e6753d9cc3</citedby><cites>FETCH-LOGICAL-c502t-494cb8a26d1955845295ef2ee2268a428bb420b22b53f5a6892dde9e6753d9cc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10295-012-1110-4$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10295-012-1110-4$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27923,27924,41487,42556,51318</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=26128005$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22395898$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kim, Ho Myeong</creatorcontrib><creatorcontrib>Lee, Yoon Gyo</creatorcontrib><creatorcontrib>Patel, Darshan H</creatorcontrib><creatorcontrib>Lee, Kwang Ho</creatorcontrib><creatorcontrib>Lee, Dae-Seok</creatorcontrib><creatorcontrib>Bae, Hyeun-Jong</creatorcontrib><title>Characteristics of bifunctional acidic endoglucanase (Cel5B) from Gloeophyllum trabeum</title><title>Journal of industrial microbiology & biotechnology</title><addtitle>J Ind Microbiol Biotechnol</addtitle><addtitle>J Ind Microbiol Biotechnol</addtitle><description>The endoglucanase (Cel5B) from the filamentous fungus Gloeophyllum trabeum was cloned and expressed without a signal peptide, and alanine residue 22 converted to glutamine in Pichia pastoris GS115. The DNA sequence of Cel5B had an open reading frame of 1,077 bp, encoding a protein of 359 amino acid residues with a molecular weight of 47 kDa. On the basis of sequence similarity, Cel5B displayed active site residues at Glu-175 and Glu-287. Both residues lost full hydrolytic activity when replaced with alanine through point mutation. The purified recombinant Cel5B showed very high specific activity, about 80- to 1,000-fold and 13- to 70-fold in comparison with other endoglucanases and cellobiohydrolase, on carboxymethylcellulose and filter paper, respectively, at pH 3.5 and 55°C. Cel5B displayed bifunctional characteristics under acidic conditions. The kinetic properties of the enzyme determined using a Lineweaver–Burk plot indicated that Cel5B is a catalytically efficient cellulolytic enzyme. These results suggest that Cel5B has high bifunctional endo- and exoglucanase activity under acidic conditions and is a good candidate for bioconversion of lignocellulose.</description><subject>alanine</subject><subject>Alternative energy sources</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Basidiomycota - enzymology</subject><subject>Basidiomycota - genetics</subject><subject>Biochemistry</subject><subject>Bioinformatics</subject><subject>Biological and medical sciences</subject><subject>Biomass</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>biotransformation</subject><subject>carboxymethylcellulose</subject><subject>Catalytic Domain</subject><subject>Cellulase</subject><subject>Cellulase - chemistry</subject><subject>Cellulase - genetics</subject><subject>Cellulase - isolation & purification</subject><subject>Cellulase - metabolism</subject><subject>Cellulose</subject><subject>cellulose 1,4-beta-cellobiosidase</subject><subject>Cellulose 1,4-beta-Cellobiosidase - genetics</subject><subject>Cellulose 1,4-beta-Cellobiosidase - metabolism</subject><subject>Cloning</subject><subject>Cloning, Molecular</subject><subject>DNA polymerase</subject><subject>endo-1,4-beta-glucanase</subject><subject>Enzymes</subject><subject>Escherichia coli - genetics</subject><subject>Fossil fuels</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>fungi</subject><subject>Genetic Engineering</subject><subject>Genetics and Molecular Biology of Industrial Organisms</subject><subject>Gloeophyllum trabeum</subject><subject>glutamine</subject><subject>Industrial Microbiology</subject><subject>Inorganic Chemistry</subject><subject>Life Sciences</subject><subject>lignocellulose</subject><subject>Microbiology</subject><subject>Microorganisms</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>molecular weight</subject><subject>Mutation</subject><subject>open reading frames</subject><subject>Peptides</subject><subject>Pichia - genetics</subject><subject>Pichia pastoris</subject><subject>point mutation</subject><subject>Protein expression</subject><subject>Proteins</subject><subject>Residues</subject><subject>Sequence Alignment</subject><subject>signal peptide</subject><subject>Yeast</subject><issn>1367-5435</issn><issn>1476-5535</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqNkk1rFTEUhoNY7If-ADc6IEK7GM33JEu91CoUXGjdhkzm5DYlM7kmM4v-e3OZqxah4CqBPOfk5H2C0EuC3xGMu_eFYKpFiwltCSG45U_QCeGdbIVg4mndM9m1gjNxjE5LucMYi66jz9AxpUwLpdUJ-rG5tdm6GXIoc3ClSb7pg18mN4c02dhYF4bgGpiGtI2Ls5Mt0JxvIIqPF43PaWyuYoK0u72PcRmbOdselvE5OvI2FnhxWM_QzafL75vP7fXXqy-bD9etE5jOLdfc9cpSORAthOKiPgc8BaBUKsup6ntOcU9pL5gXVipNhwE0yE6wQTvHztD52neX088FymzGUBzEaCdISzE1IC6lVFj_D0q0Iop0FX3zD3qXllzTWClW0-P7hmSlXE6lZPBml8No832FzN6PWf2Y6sfs_Rhea14dOi_9CMOfit9CKvD2ANjibPTZTi6Uv5wkVFWNlaMrV-rRtIX8cMTHb3-9FnmbjN1W5ebmG8WE15_BNVGc_QIRFa7I</recordid><startdate>20120701</startdate><enddate>20120701</enddate><creator>Kim, Ho Myeong</creator><creator>Lee, Yoon Gyo</creator><creator>Patel, Darshan H</creator><creator>Lee, Kwang Ho</creator><creator>Lee, Dae-Seok</creator><creator>Bae, Hyeun-Jong</creator><general>Springer-Verlag</general><general>Springer</general><general>Oxford University Press</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>3V.</scope><scope>7QL</scope><scope>7QR</scope><scope>7T7</scope><scope>7WY</scope><scope>7WZ</scope><scope>7X7</scope><scope>7XB</scope><scope>87Z</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8FL</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BEZIV</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FRNLG</scope><scope>FYUFA</scope><scope>F~G</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>HCIFZ</scope><scope>K60</scope><scope>K6~</scope><scope>K9.</scope><scope>L.-</scope><scope>LK8</scope><scope>M0C</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>P64</scope><scope>PQBIZ</scope><scope>PQBZA</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>7QO</scope></search><sort><creationdate>20120701</creationdate><title>Characteristics of bifunctional acidic endoglucanase (Cel5B) from Gloeophyllum trabeum</title><author>Kim, Ho Myeong ; Lee, Yoon Gyo ; Patel, Darshan H ; Lee, Kwang Ho ; Lee, Dae-Seok ; Bae, Hyeun-Jong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c502t-494cb8a26d1955845295ef2ee2268a428bb420b22b53f5a6892dde9e6753d9cc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>alanine</topic><topic>Alternative energy sources</topic><topic>Amino Acid Sequence</topic><topic>Amino acids</topic><topic>Basidiomycota - enzymology</topic><topic>Basidiomycota - genetics</topic><topic>Biochemistry</topic><topic>Bioinformatics</topic><topic>Biological and medical sciences</topic><topic>Biomass</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>biotransformation</topic><topic>carboxymethylcellulose</topic><topic>Catalytic Domain</topic><topic>Cellulase</topic><topic>Cellulase - chemistry</topic><topic>Cellulase - genetics</topic><topic>Cellulase - isolation & purification</topic><topic>Cellulase - metabolism</topic><topic>Cellulose</topic><topic>cellulose 1,4-beta-cellobiosidase</topic><topic>Cellulose 1,4-beta-Cellobiosidase - genetics</topic><topic>Cellulose 1,4-beta-Cellobiosidase - metabolism</topic><topic>Cloning</topic><topic>Cloning, Molecular</topic><topic>DNA polymerase</topic><topic>endo-1,4-beta-glucanase</topic><topic>Enzymes</topic><topic>Escherichia coli - genetics</topic><topic>Fossil fuels</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>fungi</topic><topic>Genetic Engineering</topic><topic>Genetics and Molecular Biology of Industrial Organisms</topic><topic>Gloeophyllum trabeum</topic><topic>glutamine</topic><topic>Industrial Microbiology</topic><topic>Inorganic Chemistry</topic><topic>Life Sciences</topic><topic>lignocellulose</topic><topic>Microbiology</topic><topic>Microorganisms</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>molecular weight</topic><topic>Mutation</topic><topic>open reading frames</topic><topic>Peptides</topic><topic>Pichia - genetics</topic><topic>Pichia pastoris</topic><topic>point mutation</topic><topic>Protein expression</topic><topic>Proteins</topic><topic>Residues</topic><topic>Sequence Alignment</topic><topic>signal peptide</topic><topic>Yeast</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kim, Ho Myeong</creatorcontrib><creatorcontrib>Lee, Yoon Gyo</creatorcontrib><creatorcontrib>Patel, Darshan H</creatorcontrib><creatorcontrib>Lee, Kwang Ho</creatorcontrib><creatorcontrib>Lee, Dae-Seok</creatorcontrib><creatorcontrib>Bae, Hyeun-Jong</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>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Chemoreception Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>ABI/INFORM Collection</collection><collection>ABI/INFORM Global (PDF only)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ABI/INFORM Global (Alumni Edition)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ABI/INFORM Collection (Alumni Edition)</collection><collection>Research Library (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Business Premium Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Business Premium Collection (Alumni)</collection><collection>Health Research Premium Collection</collection><collection>ABI/INFORM Global (Corporate)</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>Research Library Prep</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Business Collection (Alumni Edition)</collection><collection>ProQuest Business Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ABI/INFORM Professional Advanced</collection><collection>ProQuest Biological Science Collection</collection><collection>ABI/INFORM Global</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Research Library</collection><collection>Science Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Research Library (Corporate)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Business</collection><collection>ProQuest One Business (Alumni)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><jtitle>Journal of industrial microbiology & biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kim, Ho Myeong</au><au>Lee, Yoon Gyo</au><au>Patel, Darshan H</au><au>Lee, Kwang Ho</au><au>Lee, Dae-Seok</au><au>Bae, Hyeun-Jong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characteristics of bifunctional acidic endoglucanase (Cel5B) from Gloeophyllum trabeum</atitle><jtitle>Journal of industrial microbiology & biotechnology</jtitle><stitle>J Ind Microbiol Biotechnol</stitle><addtitle>J Ind Microbiol Biotechnol</addtitle><date>2012-07-01</date><risdate>2012</risdate><volume>39</volume><issue>7</issue><spage>1081</spage><epage>1089</epage><pages>1081-1089</pages><issn>1367-5435</issn><eissn>1476-5535</eissn><abstract>The endoglucanase (Cel5B) from the filamentous fungus Gloeophyllum trabeum was cloned and expressed without a signal peptide, and alanine residue 22 converted to glutamine in Pichia pastoris GS115. The DNA sequence of Cel5B had an open reading frame of 1,077 bp, encoding a protein of 359 amino acid residues with a molecular weight of 47 kDa. On the basis of sequence similarity, Cel5B displayed active site residues at Glu-175 and Glu-287. Both residues lost full hydrolytic activity when replaced with alanine through point mutation. The purified recombinant Cel5B showed very high specific activity, about 80- to 1,000-fold and 13- to 70-fold in comparison with other endoglucanases and cellobiohydrolase, on carboxymethylcellulose and filter paper, respectively, at pH 3.5 and 55°C. Cel5B displayed bifunctional characteristics under acidic conditions. The kinetic properties of the enzyme determined using a Lineweaver–Burk plot indicated that Cel5B is a catalytically efficient cellulolytic enzyme. These results suggest that Cel5B has high bifunctional endo- and exoglucanase activity under acidic conditions and is a good candidate for bioconversion of lignocellulose.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer-Verlag</pub><pmid>22395898</pmid><doi>10.1007/s10295-012-1110-4</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | alanine Alternative energy sources Amino Acid Sequence Amino acids Basidiomycota - enzymology Basidiomycota - genetics Biochemistry Bioinformatics Biological and medical sciences Biomass Biomedical and Life Sciences Biotechnology biotransformation carboxymethylcellulose Catalytic Domain Cellulase Cellulase - chemistry Cellulase - genetics Cellulase - isolation & purification Cellulase - metabolism Cellulose cellulose 1,4-beta-cellobiosidase Cellulose 1,4-beta-Cellobiosidase - genetics Cellulose 1,4-beta-Cellobiosidase - metabolism Cloning Cloning, Molecular DNA polymerase endo-1,4-beta-glucanase Enzymes Escherichia coli - genetics Fossil fuels Fundamental and applied biological sciences. Psychology fungi Genetic Engineering Genetics and Molecular Biology of Industrial Organisms Gloeophyllum trabeum glutamine Industrial Microbiology Inorganic Chemistry Life Sciences lignocellulose Microbiology Microorganisms Models, Molecular Molecular Sequence Data molecular weight Mutation open reading frames Peptides Pichia - genetics Pichia pastoris point mutation Protein expression Proteins Residues Sequence Alignment signal peptide Yeast |
| title | Characteristics of bifunctional acidic endoglucanase (Cel5B) from Gloeophyllum trabeum |
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