US 10,138,489 B2
Cyanobacterial strains capable of utilizing phosphite
Paul Gordon Roessler, San Diego, CA (US); Charles Ryan Budinoff, Fort Myers, FL (US); Songhua Zhu, Fort Myers, FL (US); Kui Wang, Fort Myers, FL (US); Heike Enke, Berlin (DE); Christian Weissert, Berlin (DE); Frank Jochem, West Palm Beach, FL (US); and Ming-De Deng, Manitowoc, WI (US)
Assigned to Algenol Biotech LLC, Fort Myers, FL (US)
Filed by Algenol Biotech LLC, Fort Myers, FL (US)
Filed on Oct. 12, 2017, as Appl. No. 15/782,404.
Claims priority of provisional application 62/410,577, filed on Oct. 20, 2016.
Prior Publication US 2018/0112225 A1, Apr. 26, 2018
Int. Cl. C12N 1/20 (2006.01); C12N 15/52 (2006.01); C07K 14/195 (2006.01); C12N 15/63 (2006.01); C12N 15/74 (2006.01); C12N 15/67 (2006.01); C12R 1/01 (2006.01); C12P 1/04 (2006.01); C12P 7/06 (2006.01)
CPC C12N 15/635 (2013.01) [C12N 1/20 (2013.01); C12N 15/52 (2013.01); C12N 15/67 (2013.01); C12N 15/74 (2013.01); C12P 1/04 (2013.01); C12P 7/065 (2013.01); C12R 1/01 (2013.01); C12Y 101/01001 (2013.01); C12Y 120/01001 (2013.01); C12Y 401/01001 (2013.01); C12N 2320/50 (2013.01); C12N 2330/51 (2013.01)] 22 Claims
1. A genetically modified cyanobacterial cell for the production of a product of interest, comprising:
a) at least one recombinant gene that encodes a heterologous phosphite dehydrogenase enzyme EC: that catalyzes the oxidation of phosphite to phosphate, wherein said enzyme has at least 85% identity to the protein sequence of the Ralstonia phosphite dehydrogenase enzyme (SEQ ID NO: 12);
b) an operon comprising at least one recombinant phosphite transporter gene encoding at least one phosphite transporter protein for transporting phosphite into the cell wherein the at least one phosphite transporter protein has at least 85% identity to a protein sequence selected from the group consisting of Cyanothece PtxA (SEQ ID NO: 19), Cyanothece PtxB (SEQ ID NO:22), Cyanothece PtxC (SEQ ID NO: 25), and Desulfotignum phosphitoxidans PtdC (SEQ ID NO: 28);
c) a knockout or knockdown of a gene encoding an endogenous protein having at least 85% identity to the repressor protein PhoU; and
d) at least one recombinant production gene encoding a polypeptide for the production of said product of interest.