ArticleApplied and environmental microbiology2012
In vitro reconstitution of the complete Clostridium thermocellum cellulosome and synergistic activity on crystalline cellulose.
Article in Applied and environmental microbiology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 60 citations in OpenAlex.
- A minimal cellulosome-like system in Cellulosilyticum lentocellum.FEBS open bio · 2026Article
- Advancing cellulose degradation through synthetic biology: engineered pathways and microbial systems for sustainable biomass conversion.Journal of animal science and biotechnology · 2026Review
- Anaerobic gut fungi as biocatalysts: metabolic and physiological analysis of anaerobic gut fungi under diverse cultivation conditions.Frontiers in microbiology · 2025Article
- Exploitation of Biodiversity in Bioeconomy: Examples, Opportunities, and Challenges.Advances in biochemical engineering/biotechnology · 2025Review
- Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold.Applied microbiology and biotechnology · 2023Article
- Current progress on engineering microbial strains and consortia for production of cellulosic butanol through consolidated bioprocessing.Microbial biotechnology · 2023Review
- Improvement of Ruminal Neutral Detergent Fiber Degradability by Obtaining and Using Exogenous Fibrolytic Enzymes from White-Rot Fungi.Animals : an open access journal from MDPI · 2022Review
- Anaerobic Fungi: Past, Present, and Future.Frontiers in microbiology · 2020Review
- Assembly of Synthetic Functional Cellulosomal Structures onto the Cell Surface of Lactobacillus plantarum, a Potent Member of the Gut Microbiome.Applied and environmental microbiology · 2018Article
- Intein-mediated assembly of tunable scaffoldins for facile synthesis of designer cellulosomes.Applied microbiology and biotechnology · 2018Article
- Optimizing the composition of a synthetic cellulosome complex for the hydrolysis of softwood pulp: identification of the enzymatic core functions and biochemical complex characterization.Biotechnology for biofuels · 2018Article
- Comparative characterization of all cellulosomal cellulases fromBiotechnology for biofuels · 2017Article
- How does cellulosome composition influence deconstruction of lignocellulosic substrates inBiotechnology for biofuels · 2017Article
- Enzymatic diversity of the Clostridium thermocellum cellulosome is crucial for the degradation of crystalline cellulose and plant biomass.Scientific reports · 2016Article
- Stoichiometric Assembly of the Cellulosome Generates Maximum Synergy for the Degradation of Crystalline Cellulose, as Revealed by In Vitro Reconstitution of the Clostridium thermocellum Cellulosome.Applied and environmental microbiology · 2015Article
- Proximity effect among cellulose-degrading enzymes displayed on the Saccharomyces cerevisiae cell surface.Applied and environmental microbiology · 2015Article
- The emergence of Clostridium thermocellum as a high utility candidate for consolidated bioprocessing applications.Frontiers in chemistry · 2014Review
- Direct glucose production from lignocellulose using Clostridium thermocellum cultures supplemented with a thermostable β-glucosidase.Biotechnology for biofuels · 2013Article
- Insertion of endocellulase catalytic domains into thermostable consensus ankyrin scaffolds: effects on stability and cellulolytic activity.Applied and environmental microbiology · 2013Article
- Functional heterologous expression of an engineered full length CipA from Clostridium thermocellum in Thermoanaerobacterium saccharolyticum.Biotechnology for biofuels · 2013Article
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Authors and funding
3 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Artificial cellulase complexes active on crystalline cellulose were reconstituted in vitro from a native mix of cellulosomal enzymes and CipA scaffoldin. Enzymes containing dockerin modules for binding to the corresponding cohesin modules were prepared from culture supernatants of a C. thermocellum cipA mutant. They were reassociated to cellulosomes via dockerin-cohesin interaction. Recombinantly produced mini-CipA proteins with one to three cohesins either with or without the carbohydrate-binding module (CBM) and the complete CipA protein were used as the cellulosomal backbone. The binding between cohesins and dockerins occurred spontaneously. The hydrolytic activity against soluble and crystalline cellulosic compounds showed that the composition of the complex does not seem to be dependent on which CipA-derived cohesin was used for reconstitution. Binding did not seem to have an obvious local preference (equal binding to Coh1 and Coh6). The synergism on crystalline cellulose increased with an increasing number of cohesins in the scaffoldin. The in vitro-formed complex showed a 12-fold synergism on the crystalline substrate (compared to the uncomplexed components). The activity of reconstituted cellulosomes with full-size CipA reached 80% of that of native cellulosomes. Complexation on the surface of nanoparticles retained the activity of protein complexes and enhanced their stability. Partial supplementation of the native cellulosome components with three selected recombinant cellulases enhanced the activity on crystalline cellulose and reached that of the native cellulosome. This opens possibilities for in vitro complex reconstitution, which is an important step toward the creation of highly efficient engineered cellulases.
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