Patent
A cyanobacterial host cell, Cyanobacterium sp., that harbors at least one recombinant gene for the production of a chemical compounds is provided, as well as vectors derived from an endogenous plasmid isolated from the cell.
1. A genetically enhanced ethanologenic Cyanobacterium sp. ABICyano1 host cell derived from wild type organism deposited in the American Type Culture Collection (ATCC) under ATCC accession number PTA-13311.
2. The host cell of claim 1 wherein said Cyanobacterium sp. ABICyano1 can tolerate salinities ranging up to about 35 practical salinity units, and
can tolerate temperatures ranging up to about 55 degrees Celsius, and
can tolerate dissolved oxygen concentrations ranging up to about 1000 μmol/L, and
can tolerate ethanol concentrations ranging up to about one percent for 16 weeks.
3. The host cell of claim 1 , wherein said host cell comprises a genetically enhanced plasmid derived from a plasmid that is endogenous to Cyanobacterium sp. ABICyano1.
4. The host cell of claim 3 , wherein said genetically enhanced plasmid comprises an origin of replication and an origin of transfer (oriVT) that has a sequence of nucleotides 5640 to 6698 of SEQ ID NO: 10.
5. The host cell of claim 3 , wherein said genetically enhanced plasmid comprises a gene encoding a replication initiation factor having an amino acid sequence identity of at least 90% to the sequence of a protein encoded by nucleotides 594 to 3779 of SEQ ID NO: 1.
6. The host cell of claim 3 , wherein said genetically enhanced plasmid comprises a heterologous alcohol dehydrogenase gene and a heterologous pyruvate decarboxylase gene, and
wherein said alcohol dehydrogenase gene is derived from Synechocystis and is operably linked to a promoter, and
wherein said pyruvate decarboxylase gene is derived from Zymomonas and is operably linked to a promoter.
7. The host cell of claim 6 , wherein said heterologous alcohol dehydrogenase gene encodes for an alcohol dehydrogenase that has an amino acid sequence identity of at least 90% to the amino acid sequence of the enzyme encoded for by nucleotides 117 to 1127 of SEQ ID NO: 15.
8. The host cell of claim 6 wherein said heterologous pyruvate decarboxylase gene encodes for a pyruvate decarboxylase having the amino acid sequence encoded by nucleotides 379 to 2085 of SEQ ID NO: 10.
9. The host cell of claim 6 wherein said promoter operably linked to an alcohol dehydrogenase gene and said promoter operably linked to a pyruvate decarboxylase gene are selected from the group consisting of PnirA, Prbc, PrpsL, PpsbA. PpetJ, PcorT, PsmtA and PpetE.
10. The host cell of claim 6 wherein said plasmid comprises # 1578 (SEQ ID NO: 15).