Elaborate cellulosome architecture of Acetivibrio cellulolyticus revealed by selective screening of cohesin–dockerin interactions
Cellulosic waste represents a significant and underutilized carbon source for the biofuel industry. Owing to the recalcitrance of crystalline cellulose to enzymatic degradation, it is necessary to design economical methods of liberating the fermentable sugars required for bioethanol production. One...
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| Format: | Article |
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PeerJ Inc.
2014-10-01
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| author | Yuval Hamberg Vered Ruimy-Israeli Bareket Dassa Yoav Barak Raphael Lamed Kate Cameron Carlos M.G.A. Fontes Edward A. Bayer Daniel B. Fried |
| author_facet | Yuval Hamberg Vered Ruimy-Israeli Bareket Dassa Yoav Barak Raphael Lamed Kate Cameron Carlos M.G.A. Fontes Edward A. Bayer Daniel B. Fried |
| author_sort | Yuval Hamberg |
| collection | DOAJ |
| description | Cellulosic waste represents a significant and underutilized carbon source for the biofuel industry. Owing to the recalcitrance of crystalline cellulose to enzymatic degradation, it is necessary to design economical methods of liberating the fermentable sugars required for bioethanol production. One route towards unlocking the potential of cellulosic waste lies in a highly complex class of molecular machines, the cellulosomes. Secreted mainly by anaerobic bacteria, cellulosomes are structurally diverse, cell surface-bound protein assemblies that can contain dozens of catalytic components. The key feature of the cellulosome is its modularity, facilitated by the ultra-high affinity cohesin–dockerin interaction. Due to the enormous number of cohesin and dockerin modules found in a typical cellulolytic organism, a major bottleneck in understanding the biology of cellulosomics is the purification of each cohesin- and dockerin-containing component, prior to analyses of their interaction. As opposed to previous approaches, the present study utilized proteins contained in unpurified whole-cell extracts. This strategy was made possible due to an experimental design that allowed for the relevant proteins to be “purified” via targeted affinity interactions as a function of the binding assay. The approach thus represents a new strategy, appropriate for future medium- to high-throughput screening of whole genomes, to determine the interactions between cohesins and dockerins. We have selected the cellulosome of Acetivibrio cellulolyticus for this work due to its exceptionally complex cellulosome systems and intriguing diversity of its cellulosomal modular components. Containing 41 cohesins and 143 dockerins, A. cellulolyticus has one of the largest number of potential cohesin–dockerin interactions of any organism, and contains unusual and novel cellulosomal features. We have surveyed a representative library of cohesin and dockerin modules spanning the cellulosome’s total cohesin and dockerin sequence diversity, emphasizing the testing of unusual and previously-unknown protein modules. The screen revealed several novel cell-bound cellulosome architectures, thus expanding on those previously known, as well as soluble cellulose systems that are not bound to the bacterial cell surface. This study sets the stage for screening the entire complement of cellulosomal components from A. cellulolyticus and other organisms with large cellulosome systems. The knowledge gained by such efforts brings us closer to understanding the exceptional catalytic abilities of cellulosomes and will allow the use of novel cellulosomal components in artificial assemblies and in enzyme cocktails for sustainable energy-related research programs. |
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| language | English |
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| publishDate | 2014-10-01 |
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| spelling | doaj.art-f9c85b8c9578449e9b2072c0d564a7f42023-12-03T00:48:07ZengPeerJ Inc.PeerJ2167-83592014-10-012e63610.7717/peerj.636636Elaborate cellulosome architecture of Acetivibrio cellulolyticus revealed by selective screening of cohesin–dockerin interactionsYuval Hamberg0Vered Ruimy-Israeli1Bareket Dassa2Yoav Barak3Raphael Lamed4Kate Cameron5Carlos M.G.A. Fontes6Edward A. Bayer7Daniel B. Fried8Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, IsraelDepartment of Biological Chemistry, The Weizmann Institute of Science, Rehovot, IsraelDepartment of Biological Chemistry, The Weizmann Institute of Science, Rehovot, IsraelDepartment of Biological Chemistry, The Weizmann Institute of Science, Rehovot, IsraelDepartment of Molecular Microbiology and Biotechnology, Tel Aviv University, Ramat Aviv, IsraelCIISA – Faculdade de Medicina Veterinária, Universidade de Lisboa, Avenida da Universidade Técnica, Lisboa, PortugalCIISA – Faculdade de Medicina Veterinária, Universidade de Lisboa, Avenida da Universidade Técnica, Lisboa, PortugalDepartment of Biological Chemistry, The Weizmann Institute of Science, Rehovot, IsraelDepartment of Biological Chemistry, The Weizmann Institute of Science, Rehovot, IsraelCellulosic waste represents a significant and underutilized carbon source for the biofuel industry. Owing to the recalcitrance of crystalline cellulose to enzymatic degradation, it is necessary to design economical methods of liberating the fermentable sugars required for bioethanol production. One route towards unlocking the potential of cellulosic waste lies in a highly complex class of molecular machines, the cellulosomes. Secreted mainly by anaerobic bacteria, cellulosomes are structurally diverse, cell surface-bound protein assemblies that can contain dozens of catalytic components. The key feature of the cellulosome is its modularity, facilitated by the ultra-high affinity cohesin–dockerin interaction. Due to the enormous number of cohesin and dockerin modules found in a typical cellulolytic organism, a major bottleneck in understanding the biology of cellulosomics is the purification of each cohesin- and dockerin-containing component, prior to analyses of their interaction. As opposed to previous approaches, the present study utilized proteins contained in unpurified whole-cell extracts. This strategy was made possible due to an experimental design that allowed for the relevant proteins to be “purified” via targeted affinity interactions as a function of the binding assay. The approach thus represents a new strategy, appropriate for future medium- to high-throughput screening of whole genomes, to determine the interactions between cohesins and dockerins. We have selected the cellulosome of Acetivibrio cellulolyticus for this work due to its exceptionally complex cellulosome systems and intriguing diversity of its cellulosomal modular components. Containing 41 cohesins and 143 dockerins, A. cellulolyticus has one of the largest number of potential cohesin–dockerin interactions of any organism, and contains unusual and novel cellulosomal features. We have surveyed a representative library of cohesin and dockerin modules spanning the cellulosome’s total cohesin and dockerin sequence diversity, emphasizing the testing of unusual and previously-unknown protein modules. The screen revealed several novel cell-bound cellulosome architectures, thus expanding on those previously known, as well as soluble cellulose systems that are not bound to the bacterial cell surface. This study sets the stage for screening the entire complement of cellulosomal components from A. cellulolyticus and other organisms with large cellulosome systems. The knowledge gained by such efforts brings us closer to understanding the exceptional catalytic abilities of cellulosomes and will allow the use of novel cellulosomal components in artificial assemblies and in enzyme cocktails for sustainable energy-related research programs.https://peerj.com/articles/636.pdfGlycoside hydrolasesCellulosomesCohesinDockerinCellulasesBiomass degradation |
| spellingShingle | Yuval Hamberg Vered Ruimy-Israeli Bareket Dassa Yoav Barak Raphael Lamed Kate Cameron Carlos M.G.A. Fontes Edward A. Bayer Daniel B. Fried Elaborate cellulosome architecture of Acetivibrio cellulolyticus revealed by selective screening of cohesin–dockerin interactions PeerJ Glycoside hydrolases Cellulosomes Cohesin Dockerin Cellulases Biomass degradation |
| title | Elaborate cellulosome architecture of Acetivibrio cellulolyticus revealed by selective screening of cohesin–dockerin interactions |
| title_full | Elaborate cellulosome architecture of Acetivibrio cellulolyticus revealed by selective screening of cohesin–dockerin interactions |
| title_fullStr | Elaborate cellulosome architecture of Acetivibrio cellulolyticus revealed by selective screening of cohesin–dockerin interactions |
| title_full_unstemmed | Elaborate cellulosome architecture of Acetivibrio cellulolyticus revealed by selective screening of cohesin–dockerin interactions |
| title_short | Elaborate cellulosome architecture of Acetivibrio cellulolyticus revealed by selective screening of cohesin–dockerin interactions |
| title_sort | elaborate cellulosome architecture of acetivibrio cellulolyticus revealed by selective screening of cohesin dockerin interactions |
| topic | Glycoside hydrolases Cellulosomes Cohesin Dockerin Cellulases Biomass degradation |
| url | https://peerj.com/articles/636.pdf |
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