Glucosyltransferase enzymes for production of glucan polymers
View Patent ↗Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan.
1. A reaction solution comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan, wherein said glucosyltransferase enzyme consists of an amino acid sequence that is at least 93% identical to SEQ ID NO:26.
2. The reaction solution of claim 1 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1,3-glucan having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100.
3. The reaction solution of claim 2 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1,3-glucan having 100% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100.
4. The reaction solution of claim 1 , further comprising a primer.
5. The reaction solution of claim 4 , wherein the primer is dextran.
6. The reaction solution of claim 4 , wherein the primer is hydrolyzed glucan.
7. The reaction solution of claim 1 , wherein said glucosyltransferase enzyme consists of an amino acid sequence that is at least 94% identical to SEQ ID NO:26.
8. The reaction solution of claim 7 , wherein said glucosyltransferase enzyme consists of an amino acid sequence that is at least 95% identical to SEQ ID NO:26.
9. The reaction solution of claim 8 , wherein said glucosyltransferase enzyme consists of an amino acid sequence that is at least 99% identical to SEQ ID NO:26.
10. The reaction solution of claim 1 , wherein a heterologous amino acid sequence of 1-300 residues is at the N-terminus and/or C-terminus of said glucosyltransferase enzyme.
11. A method for producing poly alpha-1,3-glucan comprising:
a) contacting at least water, sucrose, and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan, wherein said glucosyltransferase enzyme consists of an amino acid sequence that is at least 93% identical to SEQ ID NO:26;
whereby poly alpha-1,3-glucan is produced; and
b) optionally, isolating the poly alpha-1,3-glucan produced in step (a).
12. The method of claim 11 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1,3-glucan having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100.
13. The method of claim 12 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1,3-glucan having 100% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100.
14. The method of claim 11 , wherein step (a) further comprises contacting a primer with the water, sucrose, and glucosyltransferase enzyme.
15. The method of claim 14 , wherein the primer is dextran.
16. The method of claim 14 , wherein the primer is hydrolyzed glucan.
17. The method of claim 11 , wherein said glucosyltransferase enzyme consists of an amino acid sequence that is at least 94% identical to SEQ ID NO:26.
18. The method of claim 17 , wherein said glucosyltransferase enzyme consists of an amino acid sequence that is at least 95% identical to SEQ ID NO:26.
19. The method of claim 18 , wherein said glucosyltransferase enzyme consists of an amino acid sequence that is at least 99% identical to SEQ ID NO:26.
20. The method of claim 11 , wherein a heterologous amino acid sequence of 1-300 residues is at the N-terminus and/or C-terminus of said glucosyltransferase enzyme.