Methods for transforming eukaryotic algae
AssigneeTabletop Energy
InventorAndrew C. S. Saphire
Life of this patent
Estimating the term…
Abstract
Compositions and methods for genetically modifying algae or other photosynthetic eukaryotes to express an alcohol producing enzyme, such as pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) or any other enzymes that also synthesizes methanol, ethanol or butanol are provided. These enzymes are engineered to contain chloroplast targeting sequences which efficiently target xenotypic proteins to the chloroplast. Algae can be stably transformed with the compositions comprising chloroplast targeting constructs containing alcohol producing enzymes whereby the alcohol is produced in the chloroplast of the organism.
Claims15
A method for producing an alcohol in a transgenic eukaryotic algae cell, the method comprising the steps of:
- transforming the algae cell with a DNA construct comprising:
- a DNA sequence comprising a nucleic acid sequence encoding an alcohol producing enzyme, wherein said enzyme comprises pyruvate decarboxylase (PDC);
- a nucleic acid sequence encoding a chloroplast transit peptide (CTP) arranged to be translated in conjunction with the enzyme, wherein said CTP has at least 90% homology to SEQ ID NO:6 or SEQ ID NO:7, or at least 90% homology to a CTP from SEQ ID NOs:
- 11-20 and 22-27;
- a nucleotide regulatory element that controls expression of the enzyme and chloroplast transit peptide in the algae;
- expressing the alcohol producing enzyme in the algae cell, wherein the enzyme is targeted to the chloroplast by the encoded chloroplast transit peptide;
- and
- producing the alcohol.
A method for producing an alcohol in a transgenic eukaryotic algae cell, the method comprising the steps of:
- transforming a chloroplast genome of the algae cell with a DNA construct comprising:
- a DNA sequence comprising a nucleic acid sequence encoding an alcohol producing enzyme, wherein said enzyme comprises pyruvate decarboxylase (PDC);
- a nucleotide regulatory element that controls expression of the enzyme in the algae operably linked to the nucleic acid sequence encoding the alcohol producing enzyme;
- expressing the alcohol producing enzyme in the chloroplast;
- and
- producing the alcohol.
Dependencies are inferred from the claim text.