Hyperphotosynthetic organisms
The present disclosure identifies pathways and mechanisms to confer improved industrial fitness on engineered organisms. It also discloses engineered organisms having improved industrial fitness. Synthetic biologic engineering modules are disclosed that provide for light capture, carbon dioxide fixation, NADH production, NADPH production, thermotolerance, pH tolerance, flue gas tolerance, salt tolerance, nutrient independence and near infrared absorbance. The disclosed engineered organisms can include one or more of these modules. Also provided are methods of using the engineered organism to produce carbon-based products of interest, biomass or pharmaceutical agents.
1. An engineered cyanobacterial cell for fuel production, wherein said cell comprises a recombinant nucleic acid encoding Vitamin B 12 independent methionine synthase, wherein said Vitamin B 12 independent methionine synthase is at least 95% identical to Escherichia coli K12 MetE of SEQ ID NO: 20, and wherein said cyanobacterial cell lacks an endogenous Vitamin B 12 independent methionine synthase.
2. The engineered cyanobacterial cell of claim 1 , wherein said Vitamin B 12 independent methionine synthase is Escherichia coli K12 MetE of SEQ ID NO: 20.
3. The engineered cyanobacterial cell of claim 1 , wherein said cyanobacterial cell is a Synechococcus species.
4. The engineered cyanobacterial cell of claim 2 , wherein said cyanobacterial cell is a Synechococcus species.
5. The engineered cyanobacterial cell of claim 3 or 4 , wherein said Synechococcus species is Synechococcus sp. PCC 7002.
6. A method for conferring Vitamin B 12 independence to a cyanobacterial cell, comprising transforming said cyanobacterial cell with a nucleic acid encoding a Vitamin B 12 independent methionine synthase at least 95% identical to Escherichia coli K12 MetE of SEQ ID NO: 20, wherein said cyanobacterial cell requires exogenous Vitamin B 12 for growth prior to said transformation.
7. The method of claim 6 , wherein said Vitamin B 12 independent methionine synthase is Escherichia coli K12 MetE of SEQ ID NO: 20.
8. The method of claim 7 , wherein said cyanobacterial cell is a Synechococcus species.
9. The method of claim 6 , wherein said cyanobacterial cell is a Synechococcus species.
10. The method of claim 6 or 7 , further comprising culturing said transformed cells in media lacking Vitamin B 12 , wherein said media selects for the growth of said transformed cells.
11. The method of claim 8 or 9 , wherein said Synechococcus species is Synechococcus sp. PCC 7002.
12. A method to produce a carbon-based product of interest, comprising
culturing an engineered cyanobacterial cell in the presence of CO 2 and light under conditions suitable to produce a carbon-based product of interest, wherein said engineered cyanobacterial cell is Vitamin B 12 independent, and wherein said engineered cyanobacterial cell comprises a recombinant nucleic acid encoding a Vitamin B 12 independent methionine synthase, wherein said Vitamin B 12 independent methionine synthase is at least 95% identical to Escherichia coli K12 MetE of SEQ ID NO: 20, and wherein said cyanobacterial cell lacks an endogenous Vitamin B 12 independent methionine synthase.
13. The method of claim 12 , wherein Vitamin B 12 independent methionine synthase is Escherichia coli K12 MetE of SEQ ID NO: 20.
14. The method of claim 12 , wherein said cyanobacterial cell is a Synechococcus species.
15. The method of claim 13 , wherein said cyanobacterial cell is a Synechococcus species.
16. The method of claim 14 or 15 , wherein said Synechococcus species is Synechococcus sp. PCC 7002.