Enzymatically produced cellulose
Enzymatic reactions are disclosed herein comprising water, glucose-1-phosphate, cellodextrin, and at least one cellodextrin phosphorylase enzyme comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:2 or SEQ ID NO:6. These reactions produce a low molecular weight, insoluble cellulose with enhanced features.
1. An isolated reaction composition comprising water, glucose-1-phosphate, cellodextrin, and a cellodextrin phosphorylase enzyme comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:2 or SEQ ID NO:6, wherein the cellodextrin phosphorylase enzyme synthesizes insoluble cellulose having a weight-average degree of polymerization (DPw) of at least 15.
2. The isolated reaction composition of claim 1 , wherein said insoluble cellulose has a DPw of at least 17.
3. The isolated reaction composition of claim 1 , wherein said insoluble cellulose has a DPw of 15 to 50.
4. The isolated reaction composition of claim 1 , wherein the cellodextrin comprises cellobiose.
5. A method for producing insoluble cellulose, said method comprising:
a) contacting at least water, glucose-1-phosphate, cellodextrin, and a cellodextrin phosphorylase enzyme in an isolated reaction, wherein said cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:2 or SEQ ID NO:6,
wherein insoluble cellulose having a weight-average degree of polymerization (DPw) of at least 15 is produced; and
b) optionally, isolating the insoluble cellulose produced in step (a).
6. The method of claim 5 , wherein the insoluble cellulose produced in step (a) has a DPw of at least 17.
7. The method of claim 5 , wherein the insoluble cellulose produced in step (a) has a DPw of 15 to 50.
8. The method of claim 5 , wherein the insoluble cellulose produced in step (a) has a cellulose II crystal structure.
9. The method of claim 5 , wherein the cellodextrin comprises cellobiose.
10. The method of claim 5 , wherein said glucose-1-phosphate is provided in step (a) by providing a second reaction, wherein the products of the second reaction comprise glucose-1-phosphate.
11. The method of claim 10 , wherein the second reaction produces glucose-1-phosphate by:
(i) contacting water, inorganic phosphate, starch, a starch phosphorylase, and optionally a starch debranching enzyme;
(ii) contacting water, inorganic phosphate, sucrose, and a sucrose phosphorylase enzyme; or
(iii) contacting water, inorganic phosphate, cellulosic biomass, an endoglucanase, a cellodextrin phosphorylase, and optionally a lytic polysaccharide monooxygenase and/or a cellobiohydrolase.
12. The method of claim 10 , wherein the second reaction is provided in the same vessel in which step (a) is performed, and wherein the second reaction is performed before and/or continuously with step (a).
13. The method of claim 11 , wherein the starch debranching enzyme is a pullulanase or isoamylase.
14. The method of claim 5 , further comprising isolating the insoluble cellulose produced in step (a).
15. The method of claim 5 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.
16. The isolated reaction composition of claim 3 , wherein said insoluble cellulose has a DPw of 15 to 30.
17. The isolated reaction composition of claim 2 , wherein said insoluble cellulose has a DPw of 17 to 50.
18. The isolated reaction composition of claim 17 , wherein said insoluble cellulose has a DPw of 17 to 30.
19. The isolated reaction composition of claim 1 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.
20. The isolated reaction composition of claim 2 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.
21. The isolated reaction composition of claim 3 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.
22. The isolated reaction composition of claim 16 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.
23. The isolated reaction composition of claim 17 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.
24. The isolated reaction composition of claim 18 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.
25. The isolated reaction composition of claim 1 , wherein the cellodextrin phosphorylase enzyme is a Vibrio species cellodextrin phosphorylase enzyme or a Ruminococcus species cellodextrin phosphorylase enzyme.
26. The isolated reaction composition of claim 1 , comprised within a stainless steel vessel, plastic vessel, or glass vessel.
27. The isolated reaction composition of claim 1 , wherein the isolated reaction composition is cell-free.