Polymer grafting by polysaccharide synthases using artificial sugar acceptors
First Claim
1. A method for producing a glycosaminoglycan polymer derivative, comprising the steps of:
- providing an enzymatically active glycosaminoglycan synthase enzyme from Pasteurella multocida;
providing a synthetic, artificial acceptor for the glycosaminoglycan synthase enzyme;
combining the synthetic, artificial acceptor with the glycosaminoglycan synthase enzyme within a reaction medium, wherein the reaction medium contains at least one sugar precursor selected from the group consisting of UDP-GlcA, UDP-GlcNAc, UDP-GalNAc; and
reacting the glycosaminoglycan synthase enzyme with the synthetic, artificial acceptor to produce an oligosaccharide or polysaccharide polymer derivative.
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Abstract
The present invention relates to methodology for polymer grafting by a polysaccharide synthase and, more particularly, polymer grafting using the glycosaminoglycan synthases from Pasteurella multocida. The methodology of the present invention includes providing an enzymatically active glycosaminoglycan synthase enzyme from Pasteurella multocida, providing a synthetic, artificial acceptor for the glycosaminoglycan synthase enzyme; combining the synthetic, artificial acceptor with the glycosaminoglycan synthase enzyme within a reaction medium, wherein the reaction medium contains at least one sugar precursor selected from the group consisting of UDP-GlcA, UDP-GlcNAc, UDP-GalNAc, and reacting the glycosaminoglycan synthase enzyme with the synthetic, artificial acceptor to produce an oligosaccharide or polysaccharide polymer. The glycosaminoglycan synthase enzyme may be hyaluronan synthase, chondroitin synthase, or heparosan synthase from P. multocida, and the oligosaccharide or polysaccharide polymer may be hyaluronic acid (hyaluronan), chondroitin, heparosan, or combinations thereof.
31 Citations
19 Claims
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1. A method for producing a glycosaminoglycan polymer derivative, comprising the steps of:
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providing an enzymatically active glycosaminoglycan synthase enzyme from Pasteurella multocida;
providing a synthetic, artificial acceptor for the glycosaminoglycan synthase enzyme;
combining the synthetic, artificial acceptor with the glycosaminoglycan synthase enzyme within a reaction medium, wherein the reaction medium contains at least one sugar precursor selected from the group consisting of UDP-GlcA, UDP-GlcNAc, UDP-GalNAc; and
reacting the glycosaminoglycan synthase enzyme with the synthetic, artificial acceptor to produce an oligosaccharide or polysaccharide polymer derivative. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. A method for producing a hyaluronic acid (hyaluronan) polymer derivative, comprising the steps of:
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providing an enzymatically active hyaluronan synthase enzyme from Pasteurella multocida;
providing a synthetic, artificial acceptor for the hyaluronan synthase enzyme;
combining the synthetic, artificial acceptor with the hyaluronan synthase enzyme within a reaction medium, wherein the reaction medium contains at least one sugar precursor selected from the group consisting of UDP-GlcA and UDP-GlcNAc; and
reacting the hyaluronan synthase enzyme with the synthetic, artificial acceptor to produce an hyaluronic acid (hyaluronan) polymer derivative. - View Dependent Claims (9, 10, 11)
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12. A method for producing a chondroitin polymer derivative, comprising the steps of:
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providing an enzymatically active chondroitin synthase enzyme from Pasteurella multocida;
providing a synthetic, artificial acceptor for the chondroitin synthase enzyme;
combining the synthetic, artificial acceptor with the chondroitin synthase enzyme within a reaction medium, wherein the reaction medium contains at least one sugar precursor selected from the group consisting of UDP-GlcA and UDP-GalNAc; and
reacting the chondroitin synthase enzyme with the synthetic, artificial acceptor to produce a chondroitin polymer derivative. - View Dependent Claims (13, 14, 15)
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16. A method for producing a heparosan polymer derivative, comprising the steps of:
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providing an enzymatically active heparosan synthase enzyme from Pasteurella multocida;
providing a synthetic, artificial acceptor for the heparosan synthase enzyme;
combining the synthetic, artificial acceptor with the heparosan synthase enzyme within a reaction medium, wherein the reaction medium contains at least one sugar precursor selected from the group consisting of UDP-GlcA and UDP-GlcNAc; and
reacting the heparosan synthase enzyme with the synthetic, artificial acceptor to produce a heparosan polymer derivative. - View Dependent Claims (17, 18, 19)
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Specification