Synthesis of glucan comprising beta-1,3 glycosidic linkages with phosphorylase enzymes
AssigneeDanisco US Inc.
InventorsZheyong Yu, Slavko Kralj, Natnael Behabtu, Zhenghong Zhang, Laurie A. Howe
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Abstract
Reaction compositions are disclosed herein comprising at least water, alpha-glucose-1-phosphate (alpha-G1P), an acceptor molecule, and a beta-1,3-glucan phosphorylase enzyme. These reactions can synthesize oligosaccharides and polysaccharides with beta-1,3 glycosidic linkages. Further disclosed are methods of isolating beta-1,3-glucan.
Claims21
1-18. (canceled)
A reaction composition comprising at least water, alpha-glucose-1-phosphate (alpha-G1P), an acceptor molecule, and a beta-1,3-glucan phosphorylase enzyme comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:2, 4, 8, 10, or 12, wherein said amino acid sequence has beta-1,3-glucan phosphorylase activity,
- wherein the acceptor molecule (i) comprises one or more glucose monomeric units, (ii) does not consist of glucose, and (iii) has one or more glycosidic linkages,
- wherein the beta-1,3-glucan phosphorylase enzyme synthesizes a beta-1,3-glucan in the reaction composition and the reaction composition is cell-free.
A method for producing beta-1,3-glucan, said method comprising:
- contacting at least water, alpha-glucose-1-phosphate (alpha-G1P), an acceptor molecule, and a beta-1,3-glucan phosphorylase enzyme comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:2, 4, 8, 10, or 12, wherein said amino acid sequence has beta-1,3-glucan phosphorylase activity, wherein the acceptor molecule (i) comprises one or more glucose monomeric units, (ii) does not consist of glucose, and (iii) has one or more glycosidic linkages,
- wherein a beta-1,3-glucan is produced, and
- wherein the contacting step is performed under cell-free conditions.
Dependencies are inferred from the claim text.