Enzymatically produced cellulose

Patent metadata

Field	Value
Publication number	US-10131929-B2
Application number	US-201515532550-A
Country	US
Kind code	B2
Filing date	Dec 15, 2015
Priority date	Dec 23, 2014
Publication date	Nov 20, 2018
Grant date	Nov 20, 2018

Abstract

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.

First claim

What is claimed is:

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.

Assignees

• Du Pont
• Du Pont

Inventors

• Behabtu Natnael
• Poulose Ayrookaran J
• Yu Zheyong
• Zhang Zhenghong

Classifications

• A61P17/16

  Emollients or protectives, e.g. against radiation · CPC title

• C12Y204/01049

  Cellodextrin phosphorylase (2.4.1.49) · CPC title

• C12P19/04 Primary

  Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds · CPC title

• C12N9/1051

  Hexosyltransferases (2.4.1) · CPC title

• C08L1/02

  Cellulose; Modified cellulose · CPC title