Carbohydrate Binding Module Family 13 - Revision history

Elizabeth Ficko-Blean: /* Ligand specificities */

2025-10-31T09:23:29Z

Ligand specificities

Elizabeth Ficko-Blean: /* Functionalities */

2025-10-31T09:13:16Z

Functionalities

Elizabeth Ficko-Blean at 09:11, 31 October 2025

2025-10-31T09:11:43Z

Elizabeth Ficko-Blean: /* Ligand specificities */

2025-10-31T09:01:29Z

Ligand specificities

Elizabeth Ficko-Blean at 09:00, 31 October 2025

2025-10-31T09:00:53Z

Elizabeth Ficko-Blean: /* Structural Features */

2025-10-31T08:56:46Z

Structural Features

Elizabeth Ficko-Blean: /* Family Firsts */

2025-10-30T14:05:52Z

Family Firsts

Elizabeth Ficko-Blean: /* Functionalities */

2025-10-30T14:03:34Z

Functionalities

Elizabeth Ficko-Blean: /* Structural Features */

2025-10-30T13:44:59Z

Structural Features

Elizabeth Ficko-Blean: /* Structural Features */

2025-10-30T13:25:02Z

Structural Features

@@ Line 18: / Line 18: @@
 == Ligand specificities ==
-The first identified CBM13 domains were in plant [[Carbohydrate-binding_modules#Blurred Lines|lectins]] like the ricin toxin and agglutinin, and were found to bind galactose residues <cite>Fujimoto2013</cite>. The domains were later found to be common within many CAZymes, especially glycoside hydrolases and glycosyltransferases. Binding to galactose, lactose, and agar is common in the family <cite>Cui2018</cite>, and binding to galacto-oligosaccharides of various different linkages has been observed <cite>Ichinose2006 Jiang2012</cite>. Some structural studies have shown the CBM13 binding sites can accommodate either the non-reducing end galactose or the reducing end glucose in lactose, showing remarkable plasticity in binding preference <cite>Notenboom2002</cite>.
+The first identified CBM13 domains were in plant [[Carbohydrate-binding_modules#Blurred Lines: CBMs, Lectins and Outliers|lectins]] like the ricin toxin and agglutinin, and were found to bind galactose residues <cite>Fujimoto2013</cite>. The domains were later found to be common within many CAZymes, especially glycoside hydrolases and glycosyltransferases. Binding to galactose, lactose, and agar is common in the family <cite>Cui2018</cite>, and binding to galacto-oligosaccharides of various different linkages has been observed <cite>Ichinose2006 Jiang2012</cite>. Some structural studies have shown the CBM13 binding sites can accommodate either the non-reducing end galactose or the reducing end glucose in lactose, showing remarkable plasticity in binding preference <cite>Notenboom2002</cite>.
 There are also many examples of xylan-binding CBM13 domains <cite>Garrido2022 Hagiwara2022</cite>. Here, there is evidence of mid-chain binding to longer oligosaccharides, and that xylopentaose can bind to two binding sites simultaneously, wrapping about the CBM13 domains to do so <cite>Notenboom2002</cite>. Multiple binding sites are often functional within CBM13 domains, with the α site seemingly being the strongest <cite>Scharpf2002 Fujimoto2004</cite>. Avid binding has been demonstrated for laminarin, by a CBM13 domain found in a β-1,3-glucanase <cite>Tamashiro2012</cite>. More recently, binding to alginate has also been demonstrated <cite>Lian2024</cite> and a CBM13 domain was identified in a cycloisomaltotetraose enzyme <cite>Fujita2021</cite>.

@@ Line 31: / Line 31: @@
 Bioinformatic analysis has revealed a strong cooccurrence of CBM13 and [[GH43]] modules, with subfamily [[GH43]]_7 enzymes apparently all containing a CBM13 domain <cite>Mewis2016</cite>. In that enzyme subfamily, the α-L-arabinofuranosidase AbfB from ''Streptomyces lividans'' carries a xylan-binding CBM13 domain <cite>Vincent1997</cite>, as does an endo-β-1,4-xylanase from ''Bacteroides intestinalis'' <cite>Pereira2021</cite>. CBM13 domains are also abundant in β-agarases, found in enzyme families [[GH16]], [[GH39]], [[GH50]], [[GH86]], and [[GH118]] <cite>Veerakumar2018</cite>.
-Diverse other examples have shown that a CBM13 domain binding to the substrate of an appended glycoside hydrolase module does lead to activity potentiation through enhanced substrate [[Carbohydrate-binding_modules#Properties|proximity]] effects, such as in a [[GH16]] agarase from ''Gilvimarinus agarilyticus'' JEA5 <cite>Lee2018</cite> and a [[GH5]]_35 xylanase from  ''Paenibacillus'' sp. H2C <cite>Hagiwara2022</cite>. The enzyme endo-β-agarase I from ''Microbulbifer thermotolerans'' JAMB-A94 was engineered by fusing the [[GH16]] catalytic module to a CBM13 domain derived from the agarolytic marine bacterium ''Catenovulum agarivorans'' <cite>Cui2014</cite>, leading to a substantial increase in agar binding and hydrolysis in the fusion enzyme <cite>Alkotaini2016</cite>.
+Diverse other examples have shown that a CBM13 domain binding to the substrate of an appended glycoside hydrolase module does lead to activity potentiation through enhanced substrate [[Carbohydrate-binding_modules#Functional Roles of CBMs|proximity]] effects, such as in a [[GH16]] agarase from ''Gilvimarinus agarilyticus'' JEA5 <cite>Lee2018</cite> and a [[GH5]]_35 xylanase from  ''Paenibacillus'' sp. H2C <cite>Hagiwara2022</cite>. The enzyme endo-β-agarase I from ''Microbulbifer thermotolerans'' JAMB-A94 was engineered by fusing the [[GH16]] catalytic module to a CBM13 domain derived from the agarolytic marine bacterium ''Catenovulum agarivorans'' <cite>Cui2014</cite>, leading to a substantial increase in agar binding and hydrolysis in the fusion enzyme <cite>Alkotaini2016</cite>.
 Reaction product structure can sometimes be affected by the action of a CBM domain. In the case of the PelQ1 pectate lyase from ''Saccharobesus litoralis'', inclusion of the native CBM13 domain in the recombinant protein promoted the formation of a dimer from polygalacturonate, whereas the enzyme without CBM produced a mixture of oligosaccharides dominated by an unsaturated trimer <cite>Lian2024</cite>. The CBM13 domain from an ''Agarivorans'' sp. L11 alginate lyase apparently improves both the catalytic efficiency and heat tolerance of the enzyme, as well as increasing the proportion of disaccharides in the final reaction product mix <cite>Li2015</cite>. It is proposed that a CBM13 also contributes to controlling product length in cycloisomaltotetraose-forming CI4Tase enzymes <cite>Fujita2021</cite>.

@@ Line 31: / Line 31: @@
 Bioinformatic analysis has revealed a strong cooccurrence of CBM13 and [[GH43]] modules, with subfamily [[GH43]]_7 enzymes apparently all containing a CBM13 domain <cite>Mewis2016</cite>. In that enzyme subfamily, the α-L-arabinofuranosidase AbfB from ''Streptomyces lividans'' carries a xylan-binding CBM13 domain <cite>Vincent1997</cite>, as does an endo-β-1,4-xylanase from ''Bacteroides intestinalis'' <cite>Pereira2021</cite>. CBM13 domains are also abundant in β-agarases, found in enzyme families [[GH16]], [[GH39]], [[GH50]], [[GH86]], and [[GH118]] <cite>Veerakumar2018</cite>.
-Diverse other examples have shown that a CBM13 domain binding to the substrate of an appended glycoside hydrolase module does lead to activity potentiation through enhanced substrate proximity effects, such as in a [[GH16]] agarase from ''Gilvimarinus agarilyticus'' JEA5 <cite>Lee2018</cite> and a [[GH5]]_35 xylanase from  ''Paenibacillus'' sp. H2C <cite>Hagiwara2022</cite>. The enzyme endo-β-agarase I from ''Microbulbifer thermotolerans'' JAMB-A94 was engineered by fusing the [[GH16]] catalytic module to a CBM13 domain derived from the agarolytic marine bacterium ''Catenovulum agarivorans'' <cite>Cui2014</cite>, leading to a substantial increase in agar binding and hydrolysis in the fusion enzyme <cite>Alkotaini2016</cite>.
+Diverse other examples have shown that a CBM13 domain binding to the substrate of an appended glycoside hydrolase module does lead to activity potentiation through enhanced substrate [[Carbohydrate-binding_modules#Properties|proximity]] effects, such as in a [[GH16]] agarase from ''Gilvimarinus agarilyticus'' JEA5 <cite>Lee2018</cite> and a [[GH5]]_35 xylanase from  ''Paenibacillus'' sp. H2C <cite>Hagiwara2022</cite>. The enzyme endo-β-agarase I from ''Microbulbifer thermotolerans'' JAMB-A94 was engineered by fusing the [[GH16]] catalytic module to a CBM13 domain derived from the agarolytic marine bacterium ''Catenovulum agarivorans'' <cite>Cui2014</cite>, leading to a substantial increase in agar binding and hydrolysis in the fusion enzyme <cite>Alkotaini2016</cite>.
 Reaction product structure can sometimes be affected by the action of a CBM domain. In the case of the PelQ1 pectate lyase from ''Saccharobesus litoralis'', inclusion of the native CBM13 domain in the recombinant protein promoted the formation of a dimer from polygalacturonate, whereas the enzyme without CBM produced a mixture of oligosaccharides dominated by an unsaturated trimer <cite>Lian2024</cite>. The CBM13 domain from an ''Agarivorans'' sp. L11 alginate lyase apparently improves both the catalytic efficiency and heat tolerance of the enzyme, as well as increasing the proportion of disaccharides in the final reaction product mix <cite>Li2015</cite>. It is proposed that a CBM13 also contributes to controlling product length in cycloisomaltotetraose-forming CI4Tase enzymes <cite>Fujita2021</cite>.

@@ Line 18: / Line 18: @@
 == Ligand specificities ==
-The first identified CBM13 domains were in plant [[Carbohydrate-binding_modules#Blurred Lines|lectins]] like ricin and agglutinin, and were found to bind galactose residues <cite>Fujimoto2013</cite>. The domains were later found to be common within many CAZymes, especially glycoside hydrolases and glycosyltransferases. Binding to galactose, lactose, and agar is common in the family <cite>Cui2018</cite>, and binding to galacto-oligosaccharides of various different linkages has been observed <cite>Ichinose2006 Jiang2012</cite>. Some structural studies have shown the CBM13 binding sites can accommodate either the non-reducing end galactose or the reducing end glucose in lactose, showing remarkable plasticity in binding preference <cite>Notenboom2002</cite>.
+The first identified CBM13 domains were in plant [[Carbohydrate-binding_modules#Blurred Lines|lectins]] like the ricin toxin and agglutinin, and were found to bind galactose residues <cite>Fujimoto2013</cite>. The domains were later found to be common within many CAZymes, especially glycoside hydrolases and glycosyltransferases. Binding to galactose, lactose, and agar is common in the family <cite>Cui2018</cite>, and binding to galacto-oligosaccharides of various different linkages has been observed <cite>Ichinose2006 Jiang2012</cite>. Some structural studies have shown the CBM13 binding sites can accommodate either the non-reducing end galactose or the reducing end glucose in lactose, showing remarkable plasticity in binding preference <cite>Notenboom2002</cite>.
 There are also many examples of xylan-binding CBM13 domains <cite>Garrido2022 Hagiwara2022</cite>. Here, there is evidence of mid-chain binding to longer oligosaccharides, and that xylopentaose can bind to two binding sites simultaneously, wrapping about the CBM13 domains to do so <cite>Notenboom2002</cite>. Multiple binding sites are often functional within CBM13 domains, with the α site seemingly being the strongest <cite>Scharpf2002 Fujimoto2004</cite>. Avid binding has been demonstrated for laminarin, by a CBM13 domain found in a β-1,3-glucanase <cite>Tamashiro2012</cite>. More recently, binding to alginate has also been demonstrated <cite>Lian2024</cite> and a CBM13 domain was identified in a cycloisomaltotetraose enzyme <cite>Fujita2021</cite>.

@@ Line 18: / Line 18: @@
 == Ligand specificities ==
-The first identified CBM13 domains were in plant lectins like ricin and agglutinin, and were found to bind galactose residues <cite>Fujimoto2013</cite>. The domains were later found to be common within many CAZymes, especially glycoside hydrolases and glycosyltransferases. Binding to galactose, lactose, and agar is common in the family <cite>Cui2018</cite>, and binding to galacto-oligosaccharides of various different linkages has been observed <cite>Ichinose2006 Jiang2012</cite>. Some structural studies have shown the CBM13 binding sites can accommodate either the non-reducing end galactose or the reducing end glucose in lactose, showing remarkable plasticity in binding preference <cite>Notenboom2002</cite>.
+The first identified CBM13 domains were in plant [[Carbohydrate-binding_modules#Blurred Lines|lectins]] like ricin and agglutinin, and were found to bind galactose residues <cite>Fujimoto2013</cite>. The domains were later found to be common within many CAZymes, especially glycoside hydrolases and glycosyltransferases. Binding to galactose, lactose, and agar is common in the family <cite>Cui2018</cite>, and binding to galacto-oligosaccharides of various different linkages has been observed <cite>Ichinose2006 Jiang2012</cite>. Some structural studies have shown the CBM13 binding sites can accommodate either the non-reducing end galactose or the reducing end glucose in lactose, showing remarkable plasticity in binding preference <cite>Notenboom2002</cite>.
 There are also many examples of xylan-binding CBM13 domains <cite>Garrido2022 Hagiwara2022</cite>. Here, there is evidence of mid-chain binding to longer oligosaccharides, and that xylopentaose can bind to two binding sites simultaneously, wrapping about the CBM13 domains to do so <cite>Notenboom2002</cite>. Multiple binding sites are often functional within CBM13 domains, with the α site seemingly being the strongest <cite>Scharpf2002 Fujimoto2004</cite>. Avid binding has been demonstrated for laminarin, by a CBM13 domain found in a β-1,3-glucanase <cite>Tamashiro2012</cite>. More recently, binding to alginate has also been demonstrated <cite>Lian2024</cite> and a CBM13 domain was identified in a cycloisomaltotetraose enzyme <cite>Fujita2021</cite>.

@@ Line 24: / Line 24: @@
 == Structural Features ==
 [[File: Cbm13 overview.png|thumb|right|500px|'''Figure 1. Structure of the CBM13 domain in the multidomain protein Xyn10A from ''Streptomyces olivaceoviridis'' E-86.''' a) The overall structure with the subdomains distinctly coloured and its ligand binding tyrosine and aspartate residues of each subdomain shown as sticks (PDB accession [{{PDBlink}}1xyf 1XYF]). b) The binding site found in the α-subdomain of the CBM13 domain in complex with 2<sup>3</sup>-4-''O''-methyl-α-D-glucuronosyl-xylotriose (MeGlcUA-X3, PDB accession [{{PDBlink}}1v6x 1V6X]). c) Overlay of the subdomains showing sequence conservation within the binding sites. Single letter residue codes are coloured based on the subdomains shown in panel a) and are labelled for subdomains ⍺/β/γ, in that order.]]
-CBM13 proteins are [[Carbohydrate-binding_modules#Types|type C]] CBMs, comprising 3 internal subdomains (α, β, and γ), each approximately 40 residues in length, which fold in similar ways around a pseudo-3-fold axis, giving rise to a β-trefoil tertiary structure ('''Figure 1'''), as is also common for plant lectins. The ligand binding site in each subdomain is found in a surface exposed pocket, where binding is principally facilitated by tyrosine and aspartate residues found conserved within each subdomain. The binding sites are designated as α, β, and γ, referring to the subdomain from which they are found. The same naming system has been used for the other multivalent β-trefoil members families [[CBM42]] and [[CBM92]], which share the same modular structure as CBM13 domains.
+CBM13 proteins are [[Carbohydrate-binding_modules#Types|type B]] or [[Carbohydrate-binding_modules#Types|type C]] CBMs, comprising 3 internal subdomains (α, β, and γ), each approximately 40 residues in length, which fold in similar ways around a pseudo-3-fold axis, giving rise to a β-trefoil tertiary structure ('''Figure 1'''), as is also common for plant lectins. The ligand binding site in each subdomain is found in a surface exposed pocket, where binding is principally facilitated by tyrosine and aspartate residues found conserved within each subdomain. The binding sites are designated as α, β, and γ, referring to the subdomain from which they are found. The same naming system has been used for the other multivalent β-trefoil members families [[CBM42]] and [[CBM92]], which share the same modular structure as CBM13 domains.
 == Functionalities ==

@@ Line 36: / Line 36: @@
 == Family Firsts ==
-;First Identified
+;First Identified:The first reported characterization of a protein containing a CBM13 domain was xylanase A from ''Streptomyces lividans'' (''Sl''XynA) <cite>Morosoli1986</cite>. At that time, the CBM had not been distinguished from the xylanase domain within the gene product. Subsequent gene sequencing and sequence alignment studies demonstrated that the domain was distinct from other CBM families <cite>Dupont1998</cite> and was later categorised as CBM family 13 <cite>Tomme1998</cite>.
-:The first reported characterization of a protein containing a CBM13 domain was xylanase A from ''Streptomyces lividans'' (''Sl''XynA) <cite>Morosoli1986</cite>. At that time, the CBM had not been distinguished from the xylanase domain within the gene product. Subsequent gene sequencing and sequence alignment studies demonstrated that the domain was distinct from other CBM families <cite>Dupont1998</cite> and was later categorised as CBM family 13 <cite>Tomme1998</cite>.
+;First Structural Characterization:The structure of the first CBM13 member, defined as a carbohydrate active enzyme encoded with the CBM domain, was Xyn10A from ''Streptomyces olivaceoviridis'' E-86 (''So''XynA; <cite>Fujimoto2000</cite>; PDB: [{{PDBlink}}1xyf 1XYF]). The first structures of a CBM13 in complex with ligands were reported with ''So''Xyn10A <cite>Fujimoto2002</cite> followed very soon after by complex structures with Xyn10A from ''Streptomyces lividans'' (''Sl''Xyn10A; <cite>Notenboom2002</cite>).
 == References ==

@@ Line 29: / Line 29: @@
 Carbohydrate Binding Module family 13 has a rich history. The earliest known examples were biochemically characterised prior to their annotation as CBM13 domains. These were shown to be xylan binders increasing substrate affinity of industrial xylan-degrading enzymes <cite>Irwin1994</cite>, yet they often proved to be non-essential in xylan hydrolysing <cite>Black1995</cite> and wood pulp bleaching applications <cite>Morris1998 Leskinen2002</cite>.
-Bioinformatic analysis has revealed a strong cooccurrence of CBM13 and GH43 modules, with subfamily GH43_7 enzymes apparently all containing a CBM13 domain <cite>Mewis2016</cite>. In that enzyme subfamily, the α-L-arabinofuranosidase AbfB from ''Streptomyces lividans'' carries a xylan-binding CBM13 domain <cite>Vincent1997</cite>, as does an endo-β-1,4-xylanase from ''Bacteroides intestinalis'' <cite>Pereira2021</cite>. CBM13 domains are also abundant in β-agarases, found in enzyme families GH16, GH39, GH50, GH86, and GH118 <cite>Veerakumar2018</cite>.
+Bioinformatic analysis has revealed a strong cooccurrence of CBM13 and [[GH43]] modules, with subfamily [[GH43]]_7 enzymes apparently all containing a CBM13 domain <cite>Mewis2016</cite>. In that enzyme subfamily, the α-L-arabinofuranosidase AbfB from ''Streptomyces lividans'' carries a xylan-binding CBM13 domain <cite>Vincent1997</cite>, as does an endo-β-1,4-xylanase from ''Bacteroides intestinalis'' <cite>Pereira2021</cite>. CBM13 domains are also abundant in β-agarases, found in enzyme families [[GH16]], [[GH39]], [[GH50]], [[GH86]], and [[GH118]] <cite>Veerakumar2018</cite>.
-Diverse other examples have shown that a CBM13 domain binding to the substrate of an appended glycoside hydrolase module does lead to activity potentiation through enhanced substrate proximity effects, such as in a GH16 agarase from ''Gilvimarinus agarilyticus'' JEA5 <cite>Lee2018</cite> and a GH5_35 xylanase from  ''Paenibacillus'' sp. H2C <cite>Hagiwara2022</cite>. The enzyme endo-β-agarase I from ''Microbulbifer thermotolerans'' JAMB-A94 was engineered by fusing the GH16 catalytic module to a CBM13 domain derived from the agarolytic marine bacterium ''Catenovulum agarivorans'' <cite>Cui2014</cite>, leading to a substantial increase in agar binding and hydrolysis in the fusion enzyme <cite>Alkotaini2016</cite>.
+Diverse other examples have shown that a CBM13 domain binding to the substrate of an appended glycoside hydrolase module does lead to activity potentiation through enhanced substrate proximity effects, such as in a [[GH16]] agarase from ''Gilvimarinus agarilyticus'' JEA5 <cite>Lee2018</cite> and a [[GH5]]_35 xylanase from  ''Paenibacillus'' sp. H2C <cite>Hagiwara2022</cite>. The enzyme endo-β-agarase I from ''Microbulbifer thermotolerans'' JAMB-A94 was engineered by fusing the [[GH16]] catalytic module to a CBM13 domain derived from the agarolytic marine bacterium ''Catenovulum agarivorans'' <cite>Cui2014</cite>, leading to a substantial increase in agar binding and hydrolysis in the fusion enzyme <cite>Alkotaini2016</cite>.
 Reaction product structure can sometimes be affected by the action of a CBM domain. In the case of the PelQ1 pectate lyase from ''Saccharobesus litoralis'', inclusion of the native CBM13 domain in the recombinant protein promoted the formation of a dimer from polygalacturonate, whereas the enzyme without CBM produced a mixture of oligosaccharides dominated by an unsaturated trimer <cite>Lian2024</cite>. The CBM13 domain from an ''Agarivorans'' sp. L11 alginate lyase apparently improves both the catalytic efficiency and heat tolerance of the enzyme, as well as increasing the proportion of disaccharides in the final reaction product mix <cite>Li2015</cite>. It is proposed that a CBM13 also contributes to controlling product length in cycloisomaltotetraose-forming CI4Tase enzymes <cite>Fujita2021</cite>.