Carbohydrate Binding Module Family 32 - Revision history

Elizabeth Ficko-Blean: /* Functionalities */

2022-08-25T12:38:08Z

Functionalities

Elizabeth Ficko-Blean: /* Structural Features */

2022-08-25T12:35:25Z

Structural Features

Elizabeth Ficko-Blean: /* Structural Features */

2022-08-25T12:33:18Z

Structural Features

Elizabeth Ficko-Blean: /* Functionalities */

2022-08-25T09:35:14Z

Functionalities

Elizabeth Ficko-Blean: /* Functionalities */

2022-08-25T09:34:48Z

Functionalities

Elizabeth Ficko-Blean: /* Functionalities */

2022-08-25T09:34:24Z

Functionalities

Elizabeth Ficko-Blean: /* References */

2022-08-25T09:33:22Z

References

Elizabeth Ficko-Blean: /* Functionalities */

2022-08-25T09:26:17Z

Functionalities

Elizabeth Ficko-Blean: /* Ligand specificities */

2022-08-25T09:00:27Z

Ligand specificities

Elizabeth Ficko-Blean: /* Functionalities */

2022-08-25T08:58:45Z

Functionalities

@@ Line 29: / Line 29: @@
 CBMs, in general, are thought to [[Carbohydrate-binding modules|target pendant catalytic modules to their respective substrates]]. CtCBM32 from a ''C. thermocellum'' manannase  potentiates the hydrolytic function of the enzyme and prevents transglycosylation reactions by preferentially binding the non-reducing end of β-mannans.  Thus, the CBM32 may sterically block the access of longer manno-oligosaccharide acceptor molecules to the enzyme active site thereby enhancing hydrolysis and affecting the products of catalysis <cite>Mizutani2012</cite>.
-Alginate lyases may be mannuronate (EC 4.2.2.3), guluronate (EC 4.2.2.11) or mixed link (EC 4.2.2.-) specific lyases. The β-elimination of the 4-OH of the C5 epimers mannuronate or guluronate during alginate lyase activity results in the same 4,5-unsaturated uronate residue on the non-reducing end.  Thus, both 4,5-unsaturated ManA and GulA have the same configuration and they become identical. The ''Persicobacter sp'' CBM32 from AlyQ, a [[PL7]], specifically targets this unsaturated non-reducing end uronate rather than the individual mannuronate or guluronate residues <cite>Teh2020, Sim2017</cite>.
+Alginate lyases may be mannuronate (EC 4.2.2.3), guluronate (EC 4.2.2.11) or mixed link (EC 4.2.2.-) specific lyases. The β-elimination of the 4-OH of the C5 epimers mannuronate or guluronate during alginate lyase activity results in the same 4,5-unsaturated uronate residue on the non-reducing end.  Thus, alginate lyase released non-reducing end 4,5-unsaturated ManA and GulA have the same configuration and are identical. The ''Persicobacter sp'' CBM32 from AlyQ, a [[PL7]], specifically targets this unsaturated non-reducing end uronate rather than the individual mannuronate or guluronate residues <cite>Teh2020, Sim2017</cite>.
 CBM32s from the Gram-positive pathogen ''C. perfringens'' may well have a dual role as many of the enzymes containing CBM32s have an LPXTG motif at their C-terminal end, which generally signals for sortase-mediated anchoring to the bacterial cell wall <cite>Mazmanian1999</cite>. Thus, not only would the catalytic modules be targeted to substrate, but also the bacterium as a whole, suggesting an adhesin-like activity for these CBMs <cite>Ficko-BleanPortraitOfAnEnzyme</cite>.

@@ Line 22: / Line 22: @@
 [[File:Cbm32C.png|thumb|300px|right|'''Figure 1.'''  A secondary structure representation of the ''C. perfringens'' NagJ CpCBM32C [{{PDBlink}}2j1e 2J1e] showing the tryptophan platform, found in the terminal loop region, interacting with D-galactose from LacNac. A calcium is shown as a grey sphere. ]]
-The CBM32s share the common beta sandwich fold and have a bound structural metal ion that is most often attributed to be calcium <cite>Boraston2004</cite>. Most family members have fairly weak binding affinities (''K''<sub>a</sub> values in the mM<sup>-1</sup> and low μM<sup>-1</sup> range) <cite>Ficko-Blean2009 Ficko-Blean2006 Mizutani2012</cite>. These binding site are located at the terminal loop region within the CBM32 family. The binding sites are, in some cases, quite shallow and designed to bind monosaccharides or short oligosaccharides, making these [[Carbohydrate-binding_modules#Types|type C]] CBMs <cite>Ficko-Blean2009 Ficko-Blean2006 Boraston2007 Ficko-Blean2012</cite>.  Variability within the apical loop region within the family confers the different ligand specificities. Characteristically, there is one residue, commonly Trp but also Tyr, which provides an important hydrophobic platform for interaction with the ring of one of the sugar moieties (Figure 1) <cite>Gaskell1995 Ficko-Blean2006 Ficko-Blean2012</cite>.  In most cases, the CBM32 members interact with the saturated non-reducing end of oligosaccharides <cite>Ficko-Blean2009 Ficko-Blean2006 Boraston2007 Ficko-Blean2012 Mizutani2012</cite>. However, this is not always the case, as demonstrated by the periplasmic-binding protein from ''Y. enterocolitica'', which shares sequence identity with the CBM32 family <cite>Cantarel2009</cite> and binds polygalacturonic acid polymers <cite>Abbott2007</cite> or the CBM32 domain of the alginate lyase AlyQ, which specifically recognizes a 4,5-unsaturated uronate and not the saturated mannuronate or guluronate at the non-reducing end <cite>Teh2020</cite>.
+The CBM32s share the common beta sandwich fold and have a bound structural metal ion that is most often attributed to be calcium <cite>Boraston2004</cite>. Most family members have fairly weak binding affinities (''K''<sub>a</sub> values in the mM<sup>-1</sup> and low μM<sup>-1</sup> range) <cite>Ficko-Blean2009 Ficko-Blean2006 Mizutani2012</cite>. These binding site are located at the terminal loop region within the CBM32 family. The binding sites are, in some cases, quite shallow and designed to bind monosaccharides or short oligosaccharides, making these [[Carbohydrate-binding_modules#Types|type C]] CBMs <cite>Ficko-Blean2009 Ficko-Blean2006 Boraston2007 Ficko-Blean2012</cite>.  Variability within the apical loop region within the family confers the different ligand specificities. Characteristically, there is one residue, commonly Trp but also Tyr, which provides an important hydrophobic platform for interaction with the ring of one of the sugar moieties (Figure 1) <cite>Gaskell1995 Ficko-Blean2006 Ficko-Blean2012</cite>.  In most cases, the CBM32 members interact with the saturated non-reducing end of oligosaccharides <cite>Ficko-Blean2009 Ficko-Blean2006 Boraston2007 Ficko-Blean2012 Mizutani2012</cite>. However, this is not always the case, as demonstrated by the periplasmic-binding protein from ''Y. enterocolitica'', which shares sequence identity with the CBM32 family <cite>Cantarel2009</cite> and binds polygalacturonic acid polymers <cite>Abbott2007</cite> or the CBM32 domain of the alginate lyase AlyQ, which specifically recognizes a 4,5-unsaturated uronate at the non-reducing end <cite>Teh2020</cite>.
 Some structural examples of the complex oligosaccharide binding sites of the CBM32s can be found in the following PDB entries: [{{PDBlink}}1euu 1EUU] <cite>Gaskell1995</cite>, [{{PDBlink}}7d29 7D29]<cite>Teh2020</cite>,[{{PDBlink}}2j7m 2J7M] <cite>Ficko-Blean2006</cite>, [{{PDBlink}}4a45 4A45] <cite>Ficko-Blean2012</cite>, [{{PDBlink}}4a6o 4A6O] <cite>Ficko-Blean2012</cite>, and [{{PDBlink}}2w1u 2W1U] <cite>Ficko-Blean2009</cite>, to name just a few.  An updated list of available three-dimensional structures is available on the [http://www.cazy.org/CBM32_structure.html CBM32 page of the CAZy Database].

@@ Line 22: / Line 22: @@
 [[File:Cbm32C.png|thumb|300px|right|'''Figure 1.'''  A secondary structure representation of the ''C. perfringens'' NagJ CpCBM32C [{{PDBlink}}2j1e 2J1e] showing the tryptophan platform, found in the terminal loop region, interacting with D-galactose from LacNac. A calcium is shown as a grey sphere. ]]
-The CBM32s share the common beta sandwich fold and have a bound structural metal ion that is most often attributed to be calcium <cite>Boraston2004</cite>. Most family members have fairly weak binding affinities (''K''<sub>a</sub> values in the mM<sup>-1</sup> and low μM<sup>-1</sup> range) <cite>Ficko-Blean2009 Ficko-Blean2006 Mizutani2012</cite>. These binding site are located at the terminal loop region within the CBM32 family. The binding sites are, in some cases, quite shallow and designed to bind monosaccharides or short oligosaccharides, making these [[Carbohydrate-binding_modules#Types|type C]] CBMs <cite>Ficko-Blean2009 Ficko-Blean2006 Boraston2007 Ficko-Blean2012</cite>.  Variability within the apical loop region within the family confers the different ligand specificities. Characteristically, there is one residue, commonly Trp but also Tyr, which provides an important hydrophobic platform for interaction with the ring of one of the sugar moieties (Figure 1) <cite>Gaskell1995 Ficko-Blean2006 Ficko-Blean2012</cite>.  In most cases, the CBM32 members interact with the saturated non-reducing end of oligosaccharides <cite>Ficko-Blean2009 Ficko-Blean2006 Boraston2007 Ficko-Blean2012 Mizutani2012</cite>. However, this is not always the case, as demonstrated by the periplasmic-binding protein from ''Y. enterocolitica'', which shares sequence identity with the CBM32 family <cite>Cantarel2009</cite> and binds polygalacturonic acid polymers <cite>Abbott2007</cite> or the CBM32 domain of the alginate lyase AlyQ, which specifically recognizes a 4,5-unsaturated mannuronate and not the saturated mannuronate at the non-reducing end <cite>Teh2020</cite>.
+The CBM32s share the common beta sandwich fold and have a bound structural metal ion that is most often attributed to be calcium <cite>Boraston2004</cite>. Most family members have fairly weak binding affinities (''K''<sub>a</sub> values in the mM<sup>-1</sup> and low μM<sup>-1</sup> range) <cite>Ficko-Blean2009 Ficko-Blean2006 Mizutani2012</cite>. These binding site are located at the terminal loop region within the CBM32 family. The binding sites are, in some cases, quite shallow and designed to bind monosaccharides or short oligosaccharides, making these [[Carbohydrate-binding_modules#Types|type C]] CBMs <cite>Ficko-Blean2009 Ficko-Blean2006 Boraston2007 Ficko-Blean2012</cite>.  Variability within the apical loop region within the family confers the different ligand specificities. Characteristically, there is one residue, commonly Trp but also Tyr, which provides an important hydrophobic platform for interaction with the ring of one of the sugar moieties (Figure 1) <cite>Gaskell1995 Ficko-Blean2006 Ficko-Blean2012</cite>.  In most cases, the CBM32 members interact with the saturated non-reducing end of oligosaccharides <cite>Ficko-Blean2009 Ficko-Blean2006 Boraston2007 Ficko-Blean2012 Mizutani2012</cite>. However, this is not always the case, as demonstrated by the periplasmic-binding protein from ''Y. enterocolitica'', which shares sequence identity with the CBM32 family <cite>Cantarel2009</cite> and binds polygalacturonic acid polymers <cite>Abbott2007</cite> or the CBM32 domain of the alginate lyase AlyQ, which specifically recognizes a 4,5-unsaturated uronate and not the saturated mannuronate or guluronate at the non-reducing end <cite>Teh2020</cite>.
 Some structural examples of the complex oligosaccharide binding sites of the CBM32s can be found in the following PDB entries: [{{PDBlink}}1euu 1EUU] <cite>Gaskell1995</cite>, [{{PDBlink}}7d29 7D29]<cite>Teh2020</cite>,[{{PDBlink}}2j7m 2J7M] <cite>Ficko-Blean2006</cite>, [{{PDBlink}}4a45 4A45] <cite>Ficko-Blean2012</cite>, [{{PDBlink}}4a6o 4A6O] <cite>Ficko-Blean2012</cite>, and [{{PDBlink}}2w1u 2W1U] <cite>Ficko-Blean2009</cite>, to name just a few.  An updated list of available three-dimensional structures is available on the [http://www.cazy.org/CBM32_structure.html CBM32 page of the CAZy Database].

@@ Line 29: / Line 29: @@
 CBMs, in general, are thought to [[Carbohydrate-binding modules|target pendant catalytic modules to their respective substrates]]. CtCBM32 from a ''C. thermocellum'' manannase  potentiates the hydrolytic function of the enzyme and prevents transglycosylation reactions by preferentially binding the non-reducing end of β-mannans.  Thus, the CBM32 may sterically block the access of longer manno-oligosaccharide acceptor molecules to the enzyme active site thereby enhancing hydrolysis and affecting the products of catalysis <cite>Mizutani2012</cite>.
-Alginate lyases may be mannuronate (EC 4.2.2.3), guluronate (EC 4.2.2.11) or mixed link (EC 4.2.2.-) specific lyases. The β-elimination of the 4-OH of the C5 epimers mannuronate or guluronate during alginate lyase activity results in the same 4,5-unsaturated uronate residue on the non-reducing end.  Thus, both 4,5-unsaturated ManA and GulA have the same configuration and they become identical. The ''Persicobacter sp'' CBM32 from AlyQ a [[PL7]] specifically targets this unsaturated non-reducing end uronate rather than the individual mannuronate or guluronate residues <cite>Teh2020, Sim2017</cite>.
+Alginate lyases may be mannuronate (EC 4.2.2.3), guluronate (EC 4.2.2.11) or mixed link (EC 4.2.2.-) specific lyases. The β-elimination of the 4-OH of the C5 epimers mannuronate or guluronate during alginate lyase activity results in the same 4,5-unsaturated uronate residue on the non-reducing end.  Thus, both 4,5-unsaturated ManA and GulA have the same configuration and they become identical. The ''Persicobacter sp'' CBM32 from AlyQ, a [[PL7]], specifically targets this unsaturated non-reducing end uronate rather than the individual mannuronate or guluronate residues <cite>Teh2020, Sim2017</cite>.
 CBM32s from the Gram-positive pathogen ''C. perfringens'' may well have a dual role as many of the enzymes containing CBM32s have an LPXTG motif at their C-terminal end, which generally signals for sortase-mediated anchoring to the bacterial cell wall <cite>Mazmanian1999</cite>. Thus, not only would the catalytic modules be targeted to substrate, but also the bacterium as a whole, suggesting an adhesin-like activity for these CBMs <cite>Ficko-BleanPortraitOfAnEnzyme</cite>.

@@ Line 29: / Line 29: @@
 CBMs, in general, are thought to [[Carbohydrate-binding modules|target pendant catalytic modules to their respective substrates]]. CtCBM32 from a ''C. thermocellum'' manannase  potentiates the hydrolytic function of the enzyme and prevents transglycosylation reactions by preferentially binding the non-reducing end of β-mannans.  Thus, the CBM32 may sterically block the access of longer manno-oligosaccharide acceptor molecules to the enzyme active site thereby enhancing hydrolysis and affecting the products of catalysis <cite>Mizutani2012</cite>.
-Alginate lyases may be mannuronate (EC 4.2.2.3), guluronate (EC 4.2.2.11) or mixed link (EC 4.2.2.-) specific lyases. The β-elimination of the 4-OH of the C5 epimers mannuronate or guluronate during alginate lyase activity results in the same 4,5-unsaturated uronate residue on the non-reducing end.  Thus, both 4,5-unsaturated ManA and GulA have the same configuration and they become identical. The ''Persicobacter sp'' CBM32 from AlyQ a [PL7] specifically targets this unsaturated non-reducing end uronate rather than the individual mannuronate or guluronate residues <cite>Teh2020, Sim2017</cite>.
+Alginate lyases may be mannuronate (EC 4.2.2.3), guluronate (EC 4.2.2.11) or mixed link (EC 4.2.2.-) specific lyases. The β-elimination of the 4-OH of the C5 epimers mannuronate or guluronate during alginate lyase activity results in the same 4,5-unsaturated uronate residue on the non-reducing end.  Thus, both 4,5-unsaturated ManA and GulA have the same configuration and they become identical. The ''Persicobacter sp'' CBM32 from AlyQ a [[PL7]] specifically targets this unsaturated non-reducing end uronate rather than the individual mannuronate or guluronate residues <cite>Teh2020, Sim2017</cite>.
 CBM32s from the Gram-positive pathogen ''C. perfringens'' may well have a dual role as many of the enzymes containing CBM32s have an LPXTG motif at their C-terminal end, which generally signals for sortase-mediated anchoring to the bacterial cell wall <cite>Mazmanian1999</cite>. Thus, not only would the catalytic modules be targeted to substrate, but also the bacterium as a whole, suggesting an adhesin-like activity for these CBMs <cite>Ficko-BleanPortraitOfAnEnzyme</cite>.

@@ Line 63: / Line 63: @@
 #Ogel1994 Ögel, Z.B.  Brayford, D. and McPherson, M.J. (1994) ''Cellulose-Triggered Sporulation in the Galactose Oxidase-Producing Fungus Cladobotryum (Dactylium) Dendroides Nrrl-2903 and Its Reidentification as a Species of Fusarium.'' Mycological Research, 98(4):474-480. [http://dx.doi.org/10.1016/S0953-7562(09)81207-0 DOI:10.1016/S0953-7562(09)81207-0]
 #Suderman2017 pmid=28428153
-#Teh2020 pmid=33010888
+#Sim2017 pmid=29057942
 </biblio>
 [[Category:Carbohydrate Binding Module Families|CBM032]]

@@ Line 17: / Line 17: @@
 == Ligand specificities ==
-In 1994, the first structure of a [[carbohydrate-binding modules|carbohydrate-binding module]] of family 32, in complex with D-galactose, was determined from a fungal galactose oxidase <cite>Ito1994</cite>. Following that, a CBM32 from a multi-modular sialidase produced by ''Micromonospora viridifaciens''  was shown to demonstrate galactose and lactose binding specificity <cite>Gaskell1995 Newstead2005</cite>. A CBM32 from a ''Cellvibrio mixtus'' family 16 glycoside hydrolase binds laminarin and pustulan <cite>Centeno2006</cite>, while a CBM32 from a ''Clostridium thermocellum'' mannanase has demonstrable binding on the non-reducing end of β-mannans and β-1,4-linked mannooligosaccharides <cite>Mizutani2012</cite>. A periplasmic-binding protein, YeCBM32, from ''Yersinia enterolitica'' shares sequence identity with the CBM32 family and binds the polygalaturonic acid components of pectin <cite>Abbott2007</cite>. A CBM32 from ''Persicobacter sp'' alginate lyase AlyQ binds 4,5-unsaturated mannuronic acid on the non-reducing end of alginate oligosaccharides <cite>Teh2020</cite>.  The ''Clostridium perfringens'' CBM32s have been well studied and many of their ligand specificities were determined as follows: D-galactose, N-acetyl-D-galactosamine <cite>Boraston2007 Ficko-Blean2012 Ficko-Blean2006 </cite>, D-galactose-β-1,4-N-acetyl-D-glucosamine (LacNAc), L-fucose-α-1,2-D-galactose-β-1,4-N-acetyl-D-glucosamine (type II blood group H-trisaccharide) <cite>Ficko-Blean2006</cite>, N-acetyl-D-glucosamine, N-acetyl-D-glucosamine-β-1,3-N-acetyl-D-galactosamine,  N-acetyl-D-glucosamine-β-1,2-D-mannose,  N-acetyl-D-glucosamine-β-1,3-D-mannose (non-biological)  <cite>Ficko-Blean2009</cite>, and N-acetyl-D-glucosamine-α-1,4-D-galactose <cite>Ficko-Blean2012</cite>. Some members of CBM family 32 have exhibited a degree of substrate promiscuity; these include CpCBM32-2 from the NagH enzyme and CpCBM32C from the NagJ enzyme of ''Clostridium perfringens'' <cite>Ficko-Blean2009 Ficko-Blean2006</cite>. This CBM family has a very diverse set of ligand specificities that is reflected in the notable amino acid sequence divergence throughout the family <cite>AbbottMolBiolEvol</cite>.
+In 1994, the first structure of a [[carbohydrate-binding modules|carbohydrate-binding module]] of family 32, in complex with D-galactose, was determined from a fungal galactose oxidase <cite>Ito1994</cite>. Following that, a CBM32 from a multi-modular sialidase produced by ''Micromonospora viridifaciens''  was shown to demonstrate galactose and lactose binding specificity <cite>Gaskell1995 Newstead2005</cite>. A CBM32 from a ''Cellvibrio mixtus'' family 16 glycoside hydrolase binds laminarin and pustulan <cite>Centeno2006</cite>, while a CBM32 from a ''Clostridium thermocellum'' mannanase has demonstrable binding on the non-reducing end of β-mannans and β-1,4-linked mannooligosaccharides <cite>Mizutani2012</cite>. A periplasmic-binding protein, YeCBM32, from ''Yersinia enterolitica'' shares sequence identity with the CBM32 family and binds the polygalaturonic acid components of pectin <cite>Abbott2007</cite>. A CBM32 from ''Persicobacter sp'' alginate lyase AlyQ binds 4,5-unsaturated uronic acid on the non-reducing end of alginate oligosaccharides <cite>Teh2020</cite>.  The ''Clostridium perfringens'' CBM32s have been well studied and many of their ligand specificities were determined as follows: D-galactose, N-acetyl-D-galactosamine <cite>Boraston2007 Ficko-Blean2012 Ficko-Blean2006 </cite>, D-galactose-β-1,4-N-acetyl-D-glucosamine (LacNAc), L-fucose-α-1,2-D-galactose-β-1,4-N-acetyl-D-glucosamine (type II blood group H-trisaccharide) <cite>Ficko-Blean2006</cite>, N-acetyl-D-glucosamine, N-acetyl-D-glucosamine-β-1,3-N-acetyl-D-galactosamine,  N-acetyl-D-glucosamine-β-1,2-D-mannose,  N-acetyl-D-glucosamine-β-1,3-D-mannose (non-biological)  <cite>Ficko-Blean2009</cite>, and N-acetyl-D-glucosamine-α-1,4-D-galactose <cite>Ficko-Blean2012</cite>. Some members of CBM family 32 have exhibited a degree of substrate promiscuity; these include CpCBM32-2 from the NagH enzyme and CpCBM32C from the NagJ enzyme of ''Clostridium perfringens'' <cite>Ficko-Blean2009 Ficko-Blean2006</cite>. This CBM family has a very diverse set of ligand specificities that is reflected in the notable amino acid sequence divergence throughout the family <cite>AbbottMolBiolEvol</cite>.
 == Structural Features ==

@@ Line 30: / Line 30: @@
 The β-elimination of the 4-OH of the C5 epimers mannuronate or guluronate during alginate lyase activity results in the same 4,5-unsaturated uronate residue on the non-reducing end.  Thus, both 4,5-unsaturated ManA and GulA have the same configuration and they become identical. The ''Persicobacter sp'' CBM32 from AlyQ specifically targets this unsaturated non-reducing end uronate rather than the individual mannuronate or guluronate residues <cite>Teh2020</cite>.
 CBM32s from the Gram-positive pathogen ''C. perfringens'' may well have a dual role as many of the enzymes containing CBM32s have an LPXTG motif at their C-terminal end, which generally signals for sortase-mediated anchoring to the bacterial cell wall <cite>Mazmanian1999</cite>. Thus, not only would the catalytic modules be targeted to substrate, but also the bacterium as a whole, suggesting an adhesin-like activity for these CBMs <cite>Ficko-BleanPortraitOfAnEnzyme</cite>.
@@ Line 36: / Line 35: @@
 The types of catalytic modules that the CBM32 members are associated with vary widely and include sialidases <cite>Boraston2007</cite>, β-N-acetylglucosaminidases <cite>Rao</cite>, α-N-acetylglucosaminidases <cite>Ficko-BleanGH89</cite>, alginate lyases <cite>Teh2020</cite>, mannanases <cite>Mizutani2012</cite> and galactose oxidases <cite>Ito1994</cite>.  In enteric bacteria the CBM32 motif may occur more than once in the same enzyme and they may or may not share the same ligand specifities; this suggests the possibility of heterogenic multivalent binding events <cite>Boraston2007 Ficko-Blean2012 AbbottMolBiolEvol</cite>. Other modules that may be associated in the same enzymes are different families of CBMs, FNIII domains, and cohesin and dockerin domains <cite>Ficko-BleanPortraitOfAnEnzyme Chitayat2008 Adams2008 Ficko-Blean2012</cite>. There are now examples of CBM32s that are independent of a catalytic module, such as the YeCBM32 periplasmic-binding protein <cite>Abbott2007</cite>.
-The ligand binding loops from CpCBM32-2 from the NagH enzyme were modified to create polyol-responsive antibody mimetics for affinity purification of specific proteins <cite>Suderman2017</cite>, this is an unusual biotechnological application for a CBM and it speaks to the plasticity within the loop binding region.
+As an example of an unusual biotechnological application, the ligand binding loops from CpCBM32-2 from the NagH enzyme were modified to create polyol-responsive antibody mimetics for affinity purification of specific proteins <cite>Suderman2017</cite>. This CBM32 application speaks to the plasticity within the loop binding region.
 == Family Firsts ==