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Carbohydrate-binding module assisting glycosynthase-catalysed polymerizations.

Victoria Codera1, Harry J Gilbert2, Magda Faijes1

  • 1Laboratory of Biochemistry, Bioengineering Department, Institut Químic de Sarrià, Universitat Ramon Llull, 08017 Barcelona, Spain.

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Summary
This summary is machine-generated.

Carbohydrate-binding modules (CBMs) enhance artificial polysaccharide synthesis by glycosynthases. CBM11 influences the degree of polymerization in synthetic β-glucans, depending on polymerization rate.

Keywords:
carbohydrate binding moduledegree of polymerizationfusion proteinglycosynthasepolymerizationpolysaccharides

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Area of Science:

  • Biochemistry
  • Carbohydrate Chemistry
  • Enzymology

Background:

  • Carbohydrate-binding modules (CBMs) are protein domains that recognize and bind carbohydrates.
  • CBMs are typically found in polysaccharide-degrading enzymes (glycoside hydrolases, GHs), enhancing their catalytic activity.
  • Glycosynthases are engineered GHs that catalyze glycosidic bond formation for synthesizing complex carbohydrates.

Purpose of the Study:

  • To investigate the potential of CBMs to assist in vitro glycosynthase-catalyzed polymerization reactions for artificial polysaccharide synthesis.
  • To explore the effect of CBM11 on the synthesis of artificial 1,3-1,4-β-glucans using a specific glycosynthase.

Main Methods:

  • Utilized a glycosynthase E(134)S derived from Bacillus licheniformis lichenase for synthesizing 1,3-1,4-β-glucans.
  • Introduced CBM11, either as a standalone protein or fused to the glycosynthase, into the polymerization reaction.
  • Analyzed the effect of CBM11 on the degree of polymerization (DP) of the synthesized polysaccharides under varying polymerization rates.

Main Results:

  • The addition of CBM11 influenced the degree of polymerization (DP) of the synthesized artificial β-glucans.
  • The effect of CBM11 on DP was dependent on the rate of polymerization.
  • Both isolated CBM11 and CBM11 fused to the glycosynthase demonstrated an impact on polysaccharide chain length.

Conclusions:

  • CBMs can potentially aid in the in vitro synthesis of artificial polysaccharides via glycosynthase catalysis.
  • CBM11 modulates the DP of synthesized glycans, suggesting a role in controlling polysaccharide chain length during enzymatic synthesis.
  • The findings provide insights into the mechanism of CBMs in influencing polysaccharide synthesis and offer a strategy for producing artificial glycans with controlled properties.