IP Library Granted Patent US 10,370,690
Granted Patent B2
US 10,370,690 · App. 16/213,319 · Granted Aug 6, 2019

Microorganisms for producing glycogen and methods of using same

Inventors: Brian F. Pfleger (Madison, WI); Andrew L. Markley (Madison, WI)
Assignee: WISCONSIN ALUMNI RESEARCH FOUNDATION
C12P19/04C12N1/20C12N9/1241C12Y207/07027
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Quick Facts
Patent No.
US 10,370,690
App. No.
16/213,319
Granted
Aug 6, 2019
Kind
B2
Abstract

Recombinant microorganisms configured for increased glycogen production. The recombinant microorganisms comprise a recombinant nucleic acid configured to express or overexpress a glucose-1-phosphate adenylyltransferase. The recombinant microorganisms produce an increased amount of glycogen compared to a corresponding microorganism not comprising the recombinant nucleic acid.

Claims (23)

1. A recombinant microorganism modified with respect to a native microorganism, the recombinant microorganism comprising a recombinant nucleic acid configured to express a glucose-1-phosphate adenylyltransferase, wherein the recombinant microorganism:

is a cyanobacterium;

exhibits enhanced glucose-1-phosphate adenylyltransferase activity compared to the native microorganism; and

produces an increased amount of glycogen compared to the native microorganism while having a growth rate of at least a growth rate of the native microorganism when grown photoautotrophically in the presence of light and 10% CO 2 .

2. The recombinant microorganism of claim 1 , wherein the glucose-1-phosphate adenylyltransferase is allosterically regulated by a compound selected from the group consisting of adenosine diphosphate and adenosine monophosphate.

3. The recombinant microorganism of claim 1 , wherein the glucose-1-phosphate adenylyltransferase comprises a sequence at least 90% identical to SEQ ID NO:2.

4. The recombinant microorganism of claim 1 , wherein the glucose-1-phosphate adenylyltransferase comprises a glycine at a position corresponding to position 336 of SEQ ID NO:2.

5. The recombinant microorganism of claim 1 , wherein the nucleic acid comprises a glucose-1-phosphate adenylyltransferase coding sequence operably connected to a promoter not operably connected to the coding sequence in nature.

6. The recombinant microorganism of claim 5 , wherein the promoter is an inducible promoter.

7. The recombinant microorganism of claim 5 , wherein the promoter is a constitutive promoter.

8. The recombinant microorganism of claim 1 , wherein the recombinant microorganism exhibits a native glycogen synthase expression level.

9. The recombinant microorganism of claim 1 , wherein the recombinant microorganism exhibits native glycogen synthase activity.

10. The recombinant microorganism of claim 1 , wherein the recombinant microorganism exhibits a native 1,4-alpha-glucan-branching enzyme expression level.

11. The recombinant microorganism of claim 1 , wherein the recombinant microorganism exhibits native 1,4-alpha-glucan-branching enzyme activity.

12. The recombinant microorganism of claim 1 , wherein the recombinant microorganism is capable of producing glycogen as a mass percent of dry cell weight (DCW) in an amount of at least about 25% DCW.

13. A method of producing glycogen comprising culturing the recombinant microorganism of claim 1 .

14. The recombinant microorganism of claim 1 , wherein the recombinant microorganism exhibits a native 1,4-alpha-glucan-branching enzyme expression level and native 1,4-alpha-glucan-branching enzyme activity, and wherein the recombinant microorganism is capable of producing glycogen as a mass percent of dry cell weight (DCW) in an amount of at least about 25% DCW.

15. The recombinant microorganism of claim 14 , wherein the glucose-1-phosphate adenylyltransferase comprises a glycine at a position corresponding to position 336 of SEQ ID NO:2 and is allosterically regulated by a compound selected from the group consisting of adenosine diphosphate and adenosine monophosphate.

16. The recombinant microorganism of claim 14 , wherein the glucose-1-phosphate adenylyltransferase comprises a sequence at least 90% identical to SEQ ID NO:2.

17. The recombinant microorganism of claim 14 , wherein the nucleic acid comprises a glucose-1-phosphate adenylyltransferase coding sequence operably connected to a promoter not operably connected to the coding sequence in nature.

18. The recombinant microorganism of claim 14 , wherein the recombinant microorganism exhibits a native glycogen synthase expression level.

19. The recombinant microorganism of claim 14 , wherein the recombinant microorganism exhibits native glycogen synthase activity.

20. A method of producing glycogen comprising culturing the recombinant microorganism of claim 14 .

Assignments (2)
CONFIRMATORY LICENSE Recorded Feb 26, 2020
From: UNIVERSITY OF WISCONSIN, MADISON
To: NATIONAL SCIENCE FOUNDATION
Reel/Frame 052019/0996 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 17, 2019
From: PFLEGER, BRIAN; MARKLEY, ANDREW
To: WISCONSIN ALUMNI RESEARCH FOUNDATION
Reel/Frame 048049/0936 →
Continuity (3)
Continuation 15366676 · Dec 1, 2016
Provisional Application 62262106 · Dec 2, 2015
Related Publication 20190093138A1 · Mar 28, 2019