Microorganisms for producing glycogen and methods of using same
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.
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 .