Purification and characterization of an endo-1,3-beta-glucanase from Aspergillus fumigatus
An endo-1,3-beta-glucanase was purified from a cell wall autolysate of Aspergillus fumigatus. This beta-glucanase activity was associated with a glycosylated 74-kDa protein. Using a sensitive colorimetric assay and a high-performance anion-exchange chromatography with a pulsed electrochemical detect...
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| Veröffentlicht in: | European Journal of Biochemistry 1997-01, Vol.243 (1/2), p.315-321 |
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| creator | Fontaine, T Hartland, R.P Beauvais, A Diaquin, M Latge, J.P |
| description | An endo-1,3-beta-glucanase was purified from a cell wall autolysate of Aspergillus fumigatus. This beta-glucanase activity was associated with a glycosylated 74-kDa protein. Using a sensitive colorimetric assay and a high-performance anion-exchange chromatography with a pulsed electrochemical detector for product analysis, it was shown that the endoglucanase hydrolysed exclusively linear 1,3-beta-glucan chains, had an optimum pH of 7.0 and an optimum temperature of 60 degrees C. A substrate kinetic study gave a Km value of 0.3 mg/ml for soluble (laminarin and laminari-oligosaccharides) and 1.18 mg/ml for insoluble (curdlan) 1,3-beta-glucan. Laminari-oligosaccharide degradation, analysed by HPLC, showed that the endoglucanase bind to the substrate at several positions and suggested that the active site of the enzyme recognized five glucose units linked by a 1,3-beta bond. The association of the present endo-1,3-beta-glucanase with the cell wall of A. fumigatus suggests a putative role for this enzyme during cell-wall morphogenesis. |
| doi_str_mv | 10.1111/j.1432-1033.1997.0315a.x |
| format | Article |
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This beta-glucanase activity was associated with a glycosylated 74-kDa protein. Using a sensitive colorimetric assay and a high-performance anion-exchange chromatography with a pulsed electrochemical detector for product analysis, it was shown that the endoglucanase hydrolysed exclusively linear 1,3-beta-glucan chains, had an optimum pH of 7.0 and an optimum temperature of 60 degrees C. A substrate kinetic study gave a Km value of 0.3 mg/ml for soluble (laminarin and laminari-oligosaccharides) and 1.18 mg/ml for insoluble (curdlan) 1,3-beta-glucan. Laminari-oligosaccharide degradation, analysed by HPLC, showed that the endoglucanase bind to the substrate at several positions and suggested that the active site of the enzyme recognized five glucose units linked by a 1,3-beta bond. 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This beta-glucanase activity was associated with a glycosylated 74-kDa protein. Using a sensitive colorimetric assay and a high-performance anion-exchange chromatography with a pulsed electrochemical detector for product analysis, it was shown that the endoglucanase hydrolysed exclusively linear 1,3-beta-glucan chains, had an optimum pH of 7.0 and an optimum temperature of 60 degrees C. A substrate kinetic study gave a Km value of 0.3 mg/ml for soluble (laminarin and laminari-oligosaccharides) and 1.18 mg/ml for insoluble (curdlan) 1,3-beta-glucan. Laminari-oligosaccharide degradation, analysed by HPLC, showed that the endoglucanase bind to the substrate at several positions and suggested that the active site of the enzyme recognized five glucose units linked by a 1,3-beta bond. The association of the present endo-1,3-beta-glucanase with the cell wall of A. fumigatus suggests a putative role for this enzyme during cell-wall morphogenesis.</description><subject>Aspergillus fumigatus</subject><subject>Aspergillus fumigatus - enzymology</subject><subject>beta-Glucans</subject><subject>Cell Wall</subject><subject>Cell Wall - enzymology</subject><subject>endo‐1,3‐β‐glucanase</subject><subject>filamentous fungus</subject><subject>Fungal Proteins</subject><subject>Fungal Proteins - isolation & purification</subject><subject>Glucan Endo-1,3-beta-D-Glucosidase</subject><subject>Glucan Endo-1,3-beta-D-Glucosidase - antagonists & inhibitors</subject><subject>Glucan Endo-1,3-beta-D-Glucosidase - isolation & purification</subject><subject>Glucans</subject><subject>Glucans - metabolism</subject><subject>Glycosylation</subject><subject>Hydrogen-Ion Concentration</subject><subject>Life Sciences</subject><subject>Membrane Glycoproteins</subject><subject>Membrane Glycoproteins - isolation & purification</subject><subject>Molecular Weight</subject><subject>plant biochemistry</subject><subject>plant physiology</subject><subject>Polysaccharides</subject><subject>Polysaccharides - metabolism</subject><subject>Substrate Specificity</subject><subject>Temperature</subject><issn>0014-2956</issn><issn>1432-1033</issn><issn>1432-1327</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU1v1DAQhi0EKkvhJyByQhxIGMffF6RQtRRpJZBKL1wsx7G3XuVjsRNo-fUkZLVHwBdbM8-8Y-lBKMNQ4Pm82xeYkjLHQEiBlRIFEMxMcf8IbU6Nx2gDgGleKsafomcp7QGAKy7O0JkCAoLRDfr2ZYrBB2vGMPSZ6ZvM3plo7Ohi-LUWBz_XM9c3Q47fkrx2o8l37WRNb5LLfBy6rEoHF3ehbaeU-akLOzNO6Tl64k2b3IvjfY5ury6_Xlzn288fP11U29wyiquc-IZ51XBS25LYWnBFsBUN9ZwCKGud91aUxjDnmBVMGgdQ1jW3tJYKlzU5R_mae2dafYihM_FBDybo62qrDyaNbooaqKBSyfIHnvnXK3-Iw_fJpVF3IVnXtqZ3w5S0kBIzgfk_QcypFIqwGXzzd7BkQnKQZMmUK2rjkFJ0_vRjDHpRq_d6MagXg3pRq_-o1ffz6MvjlqnuXHMaPLqc--_X_s_Quof_ztVXlx9u5mc1B7xaA7wZtNnFkPTtTQmYQCk5ZaDIb20Ku9I</recordid><startdate>199701</startdate><enddate>199701</enddate><creator>Fontaine, T</creator><creator>Hartland, R.P</creator><creator>Beauvais, A</creator><creator>Diaquin, M</creator><creator>Latge, J.P</creator><general>Blackwell Science Ltd</general><general>Wiley</general><scope>FBQ</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>M7N</scope><scope>7X8</scope><scope>1XC</scope><orcidid>https://orcid.org/0000-0002-8184-789X</orcidid></search><sort><creationdate>199701</creationdate><title>Purification and characterization of an endo-1,3-beta-glucanase from Aspergillus fumigatus</title><author>Fontaine, T ; Hartland, R.P ; Beauvais, A ; Diaquin, M ; Latge, J.P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c541A-3fd5f9d63bc23cb76931c7d4f64009cceffc72aa5ee5c758ae002bb6c4b8912b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Aspergillus fumigatus</topic><topic>Aspergillus fumigatus - enzymology</topic><topic>beta-Glucans</topic><topic>Cell Wall</topic><topic>Cell Wall - enzymology</topic><topic>endo‐1,3‐β‐glucanase</topic><topic>filamentous fungus</topic><topic>Fungal Proteins</topic><topic>Fungal Proteins - isolation & purification</topic><topic>Glucan Endo-1,3-beta-D-Glucosidase</topic><topic>Glucan Endo-1,3-beta-D-Glucosidase - antagonists & inhibitors</topic><topic>Glucan Endo-1,3-beta-D-Glucosidase - isolation & purification</topic><topic>Glucans</topic><topic>Glucans - metabolism</topic><topic>Glycosylation</topic><topic>Hydrogen-Ion Concentration</topic><topic>Life Sciences</topic><topic>Membrane Glycoproteins</topic><topic>Membrane Glycoproteins - isolation & purification</topic><topic>Molecular Weight</topic><topic>plant biochemistry</topic><topic>plant physiology</topic><topic>Polysaccharides</topic><topic>Polysaccharides - metabolism</topic><topic>Substrate Specificity</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fontaine, T</creatorcontrib><creatorcontrib>Hartland, R.P</creatorcontrib><creatorcontrib>Beauvais, A</creatorcontrib><creatorcontrib>Diaquin, M</creatorcontrib><creatorcontrib>Latge, J.P</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><jtitle>European Journal of Biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Fontaine, T</au><au>Hartland, R.P</au><au>Beauvais, A</au><au>Diaquin, M</au><au>Latge, J.P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification and characterization of an endo-1,3-beta-glucanase from Aspergillus fumigatus</atitle><jtitle>European Journal of Biochemistry</jtitle><addtitle>Eur J Biochem</addtitle><date>1997-01</date><risdate>1997</risdate><volume>243</volume><issue>1/2</issue><spage>315</spage><epage>321</epage><pages>315-321</pages><issn>0014-2956</issn><eissn>1432-1033</eissn><eissn>1432-1327</eissn><abstract>An endo-1,3-beta-glucanase was purified from a cell wall autolysate of Aspergillus fumigatus. This beta-glucanase activity was associated with a glycosylated 74-kDa protein. Using a sensitive colorimetric assay and a high-performance anion-exchange chromatography with a pulsed electrochemical detector for product analysis, it was shown that the endoglucanase hydrolysed exclusively linear 1,3-beta-glucan chains, had an optimum pH of 7.0 and an optimum temperature of 60 degrees C. A substrate kinetic study gave a Km value of 0.3 mg/ml for soluble (laminarin and laminari-oligosaccharides) and 1.18 mg/ml for insoluble (curdlan) 1,3-beta-glucan. Laminari-oligosaccharide degradation, analysed by HPLC, showed that the endoglucanase bind to the substrate at several positions and suggested that the active site of the enzyme recognized five glucose units linked by a 1,3-beta bond. The association of the present endo-1,3-beta-glucanase with the cell wall of A. fumigatus suggests a putative role for this enzyme during cell-wall morphogenesis.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>9030754</pmid><doi>10.1111/j.1432-1033.1997.0315a.x</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0002-8184-789X</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Aspergillus fumigatus Aspergillus fumigatus - enzymology beta-Glucans Cell Wall Cell Wall - enzymology endo‐1,3‐β‐glucanase filamentous fungus Fungal Proteins Fungal Proteins - isolation & purification Glucan Endo-1,3-beta-D-Glucosidase Glucan Endo-1,3-beta-D-Glucosidase - antagonists & inhibitors Glucan Endo-1,3-beta-D-Glucosidase - isolation & purification Glucans Glucans - metabolism Glycosylation Hydrogen-Ion Concentration Life Sciences Membrane Glycoproteins Membrane Glycoproteins - isolation & purification Molecular Weight plant biochemistry plant physiology Polysaccharides Polysaccharides - metabolism Substrate Specificity Temperature |
| title | Purification and characterization of an endo-1,3-beta-glucanase from Aspergillus fumigatus |
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