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Carbohydrate Binding Module Family 58

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Ligand specificities

Only a single CBM58 family member has been characterized, the founding member from in the neopullulanase SusG of the human gut symbiont Bacteroides thetaiotaomicron [1]. The crystal structure of SusG featuring CBM58 revealed binding to maltoheptaose as well as acarbose. Isothermal titration calorimetry as well as affinity PAGE demonstrates that the CBM58 of SusG binds maltoheptaose, α-cyclodextrin and amylopectin. Based upon these data as reported by Koropatkin[1], the CBM58 family is believed to bind exclusively to α1,4-linked glucan structures in starch, with no apparent recognition or affinity for regions displaying α1,6- branching.

Structural Features

The crystal structure of CBM58 from SusG of B. thetaiotaomicron displays a canonical β-sandwich fold, with a single binding site accomodated by the loops connecting β3 and β4, connecting β6 and β7, and β7 and β8 [1]. These loops fold over one of the β-sheets. Because CBM58 recognizes internal regions of the polypeptide chain, it can be classified as a type B CBM [2]. Like many starch-specific CBMs [3], the helical α -glucan at CBM58 in SusG is cradled by aromatic stacking interactions with two aromatic residues, W287 and W299, as well as hydrogen bonding interactions with K304, N330 and Y260 that aid in positioning the adjacent glucose residues stacking with the two tryptophans [1].

A unique facet of the CBM58 of SusG is its position in the middle of the polypeptide chain, occupying residues 216-335 of the protein. This domain is inserted between α3 and β3 of the catalytic domain the, and essentially expands the typically small B domain of GH13 enzymes. In SusG, two short linker sequences from residues 212-217 and 334-338 span about 12Å between the A domain and the CBM58 have B-factors and an amino acid sequence imply this domain is held in a fixed position, without inherent flexibility. There are only 14 other members of the CBM58 family, and they all reside within the Bacteroidetes phylum and include similar GH13 enzymes from isolates of Alistipes finegoldii, Alistipes shahii, Bacteroides dorei, Bacteroides eggerthii, and Bacteroides vulgatus. A sequence alignment of all 15 GH13 enzymes possessing a CBM58 reveals that the location of this domain is invariant as an extension of the B domain.


  0  0  1  156  910  University of Michigan Medical School  14  5  1061  14.0

CBM58 is only found within GH13 enzymes, and the conserved placement of the domain within the B. thetaiotaomicron SusG and like enzymes is somewhat unique as it interrupts the fold of the catalytic domain. In the structure of SusG, the starch-binding face of CBM58 is located 45Å and on the opposite face of the protein from the catalytic cleft. A mutant of SusG in which the CBM58 has been deleted retains catalytic activity against small substrates such as PNP-maltohexaose similar to the wild-type enzyme. The CBM-less SusG enzyme is 2-3 fold more active on autoclaved soluble starch such as maize amylopectin, but only retains ~40% of the wild-type activity on insoluble corn starch. B. thetaiotaomicron expressing only an allele for the CBM-less SusG grow the same as wild-type cells on autoclaved soluble starches.

While the CBM58 of SusG has not been used in any novel applications, this CBM was replaced with the Halo-tag® protein for single-molecule fluorescence imaging of the SusG protein in live B. thetaiotaomicron cultured on glucose and starch.

Family Firsts

First Identified
The CBM58 domain was identified during the biochemical and structural characterization of SusG from B. thetaiotaomicron.
First Structural Characterization
The crystal structure of the GH13 enzyme SusG with maltoheptaose revealed an extended B domain as CBM58 that recognizes starch and maltooligosaccharides.


  1. Koropatkin NM and Smith TJ. (2010) SusG: a unique cell-membrane-associated alpha-amylase from a prominent human gut symbiont targets complex starch molecules. Structure. 18, 200-15. DOI:10.1016/j.str.2009.12.010 | PubMed ID:20159465 | HubMed [Koropatkin2010]
  2. Gilbert HJ, Knox JP, and Boraston AB. (2013) Advances in understanding the molecular basis of plant cell wall polysaccharide recognition by carbohydrate-binding modules. Curr Opin Struct Biol. 23, 669-77. DOI:10.1016/ | PubMed ID:23769966 | HubMed [Gilbert2013]
  3. Machovic M and Janecek S. (2006) Starch-binding domains in the post-genome era. Cell Mol Life Sci. 63, 2710-24. DOI:10.1007/s00018-006-6246-9 | PubMed ID:17013558 | HubMed [Machovic2006]
  4. Davies, G.J. and Sinnott, M.L. (2008) Sorting the diverse: the sequence-based classifications of carbohydrate-active enzymes. The Biochemist, vol. 30, no. 4., pp. 26-32. Download PDF version.
  5. Boraston AB, Bolam DN, Gilbert HJ, and Davies GJ. (2004) Carbohydrate-binding modules: fine-tuning polysaccharide recognition. Biochem J. 382, 769-81. DOI:10.1042/BJ20040892 | PubMed ID:15214846 | HubMed [Boraston2004]
  6. Hashimoto H (2006) Recent structural studies of carbohydrate-binding modules. Cell Mol Life Sci. 63, 2954-67. DOI:10.1007/s00018-006-6195-3 | PubMed ID:17131061 | HubMed [Hashimoto2006]
  7. Shoseyov O, Shani Z, and Levy I. (2006) Carbohydrate binding modules: biochemical properties and novel applications. Microbiol Mol Biol Rev. 70, 283-95. DOI:10.1128/MMBR.00028-05 | PubMed ID:16760304 | HubMed [Shoseyov2006]
  8. Guillén D, Sánchez S, and Rodríguez-Sanoja R. (2010) Carbohydrate-binding domains: multiplicity of biological roles. Appl Microbiol Biotechnol. 85, 1241-9. DOI:10.1007/s00253-009-2331-y | PubMed ID:19908036 | HubMed [Guillen2010]
All Medline abstracts: PubMed | HubMed