ArticleProceedings of the National Academy of Sciences of the United States of America2012
How nature can exploit nonspecific catalytic and carbohydrate binding modules to create enzymatic specificity.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 68 papers.
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Who cites it
68 citing papers in PubMed, 169 citations in OpenAlex.
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- Gatekeeping Dietary Fiber: The Role of Carbohydrate-Binding Modules in the Human Gut.Journal of microbiology and biotechnology · 2026Review
- Enhanced fructooligosaccharides synthesis by engineered Trichoderma atroviride β-fructofuranosidase.Applied microbiology and biotechnology · 2026Article
- Organic matter degradation by oceanic fungi differs between polar and non-polar waters.Nature communications · 2025Article
- Genome-Wide Identification and Analysis of Carbohydrate-Binding Modules inInternational journal of molecular sciences · 2025Article
- Characterization of a Fructan Exohydrolase from the Gut BacteriumJournal of applied glycoscience · 2025Article
- In vitro and ex vivo metabolism of chemically diverse fructans by bovine rumen Bifidobacterium and Lactobacillus species.Animal microbiome · 2024Article
- An Exopolysaccharide from the CyanobacteriumACS omega · 2024Review
- Functional studies on tandem carbohydrate-binding modules of a multimodular enzyme possessing two catalytic domains.Applied and environmental microbiology · 2024Article
- Expanding the horizons of levan: from microbial biosynthesis to applications and advanced detection methods.World journal of microbiology & biotechnology · 2024Review
- Three-component systems represent a common pathway for extracytoplasmic addition of pentofuranose sugars into bacterial glycans.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Article
- Identification of a novel xanthan-binding module of a multi-modularFrontiers in microbiology · 2024Article
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- Combinatorial metabolic engineering enables high yield production of α-arbutin from sucrose by biocatalysis.Applied microbiology and biotechnology · 2023Article
- New carbohydrate binding domains identified by phage display based functional metagenomic screens of human gut microbiota.Communications biology · 2023Article
- Exploring Host-Binding Machineries of Mycobacteriophages with AlphaFold2.Journal of virology · 2023Article
- Functional identification of two novel carbohydrate-binding modules of glucuronoxylanase CrXyl30 and their contribution to the lignocellulose saccharification.Biotechnology for biofuels and bioproducts · 2023Article
- Enhancement of catalytic activity and alkaline stability of cellobiohydrolase by structure-based protein engineering.3 Biotech · 2022Article
8 more citing papers are in PubMed but not listed here.
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Authors and funding
11 authors at 6 institutions in 4 countries.
Funding
Abstract
Noncatalytic carbohydrate binding modules (CBMs) are components of glycoside hydrolases that attack generally inaccessible substrates. CBMs mediate a two- to fivefold elevation in the activity of endo-acting enzymes, likely through increasing the concentration of the appended enzymes in the vicinity of the substrate. The function of CBMs appended to exo-acting glycoside hydrolases is unclear because their typical endo-binding mode would not fulfill a targeting role. Here we show that the Bacillus subtilis exo-acting β-fructosidase SacC, which specifically hydrolyses levan, contains the founding member of CBM family 66 (CBM66). The SacC-derived CBM66 (BsCBM66) targets the terminal fructosides of the major fructans found in nature. The crystal structure of BsCBM66 in complex with ligands reveals extensive interactions with the terminal fructose moiety (Fru-3) of levantriose but only limited hydrophobic contacts with Fru-2, explaining why the CBM displays broad specificity. Removal of BsCBM66 from SacC results in a ~100-fold reduction in activity against levan. The truncated enzyme functions as a nonspecific β-fructosidase displaying similar activity against β-2,1- and β-2,6-linked fructans and their respective fructooligosaccharides. Conversely, appending BsCBM66 to BT3082, a nonspecific β-fructosidase from Bacteroides thetaiotaomicron, confers exolevanase activity on the enzyme. We propose that BsCBM66 confers specificity for levan, a branched fructan, through an "avidity" mechanism in which the CBM and the catalytic module target the termini of different branches of the same polysaccharide molecule. This report identifies a unique mechanism by which CBMs modulate enzyme function, and shows how specificity can be tailored by integrating nonspecific catalytic and binding modules into a single enzyme.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.