A thermostable GH45 endoglucanase from yeast: impact of its atypical multimodularity on activity
<p>Abstract</p> <p>Background</p> <p>The gene encoding an atypical multi-modular glycoside hydrolase family 45 endoglucanase bearing five different family 1 carbohydrate binding modules (CBM1), designated <it>Pp</it>Cel45A, was identified in the <it>Pi...
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| Format: | Article |
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BMC
2011-12-01
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| Series: | Microbial Cell Factories |
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| Online Access: | http://www.microbialcellfactories.com/content/10/1/103 |
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| author | Couturier Marie Feliu Julia Haon Mireille Navarro David Lesage-Meessen Laurence Coutinho Pedro M Berrin Jean-Guy |
| author_facet | Couturier Marie Feliu Julia Haon Mireille Navarro David Lesage-Meessen Laurence Coutinho Pedro M Berrin Jean-Guy |
| author_sort | Couturier Marie |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The gene encoding an atypical multi-modular glycoside hydrolase family 45 endoglucanase bearing five different family 1 carbohydrate binding modules (CBM1), designated <it>Pp</it>Cel45A, was identified in the <it>Pichia pastoris </it>GS115 genome.</p> <p>Results</p> <p><it>Pp</it>Cel45A (full-length open reading frame), and three derived constructs comprising (i) the catalytic module with its proximal CBM1, (ii) the catalytic module only, and (iii) the five CBM1 modules without catalytic module, were successfully expressed to high yields (up to 2 grams per litre of culture) in <it>P. pastoris </it>X33. Although the constructs containing the catalytic module displayed similar activities towards a range of glucans, comparison of their biochemical characteristics revealed striking differences. We observed a high thermostability of <it>Pp</it>Cel45A (Half life time of 6 h at 80°C), which decreased with the removal of CBMs and glycosylated linkers. However, both binding to crystalline cellulose and hydrolysis of crystalline cellulose and cellohexaose were substantially boosted by the presence of one CBM rather than five.</p> <p>Conclusions</p> <p>The present study has revealed the specific features of the first characterized endo β-1,4 glucanase from yeast, whose thermostability is promising for biotechnological applications related to the saccharification of lignocellulosic biomass such as consolidated bioprocessing.</p> |
| first_indexed | 2024-12-12T10:15:14Z |
| format | Article |
| id | doaj.art-d67e5cf0eb86406596197f5962ba5704 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-12T10:15:14Z |
| publishDate | 2011-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-d67e5cf0eb86406596197f5962ba57042022-12-22T00:27:41ZengBMCMicrobial Cell Factories1475-28592011-12-0110110310.1186/1475-2859-10-103A thermostable GH45 endoglucanase from yeast: impact of its atypical multimodularity on activityCouturier MarieFeliu JuliaHaon MireilleNavarro DavidLesage-Meessen LaurenceCoutinho Pedro MBerrin Jean-Guy<p>Abstract</p> <p>Background</p> <p>The gene encoding an atypical multi-modular glycoside hydrolase family 45 endoglucanase bearing five different family 1 carbohydrate binding modules (CBM1), designated <it>Pp</it>Cel45A, was identified in the <it>Pichia pastoris </it>GS115 genome.</p> <p>Results</p> <p><it>Pp</it>Cel45A (full-length open reading frame), and three derived constructs comprising (i) the catalytic module with its proximal CBM1, (ii) the catalytic module only, and (iii) the five CBM1 modules without catalytic module, were successfully expressed to high yields (up to 2 grams per litre of culture) in <it>P. pastoris </it>X33. Although the constructs containing the catalytic module displayed similar activities towards a range of glucans, comparison of their biochemical characteristics revealed striking differences. We observed a high thermostability of <it>Pp</it>Cel45A (Half life time of 6 h at 80°C), which decreased with the removal of CBMs and glycosylated linkers. However, both binding to crystalline cellulose and hydrolysis of crystalline cellulose and cellohexaose were substantially boosted by the presence of one CBM rather than five.</p> <p>Conclusions</p> <p>The present study has revealed the specific features of the first characterized endo β-1,4 glucanase from yeast, whose thermostability is promising for biotechnological applications related to the saccharification of lignocellulosic biomass such as consolidated bioprocessing.</p>http://www.microbialcellfactories.com/content/10/1/103biomasscrystalline cellulosecellulase<it>Pichia pastoris</it>thermostable GH45 endoglucanase |
| spellingShingle | Couturier Marie Feliu Julia Haon Mireille Navarro David Lesage-Meessen Laurence Coutinho Pedro M Berrin Jean-Guy A thermostable GH45 endoglucanase from yeast: impact of its atypical multimodularity on activity Microbial Cell Factories biomass crystalline cellulose cellulase <it>Pichia pastoris</it> thermostable GH45 endoglucanase |
| title | A thermostable GH45 endoglucanase from yeast: impact of its atypical multimodularity on activity |
| title_full | A thermostable GH45 endoglucanase from yeast: impact of its atypical multimodularity on activity |
| title_fullStr | A thermostable GH45 endoglucanase from yeast: impact of its atypical multimodularity on activity |
| title_full_unstemmed | A thermostable GH45 endoglucanase from yeast: impact of its atypical multimodularity on activity |
| title_short | A thermostable GH45 endoglucanase from yeast: impact of its atypical multimodularity on activity |
| title_sort | thermostable gh45 endoglucanase from yeast impact of its atypical multimodularity on activity |
| topic | biomass crystalline cellulose cellulase <it>Pichia pastoris</it> thermostable GH45 endoglucanase |
| url | http://www.microbialcellfactories.com/content/10/1/103 |
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