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US 9,493,520 B2Granted
Fatty acid desaturases and elongases and uses thereof
AssigneeBASF Plant Science GmbH
InventorsJörg Bauer, Johnathan A. Napier, Olga Sayanova
Life of this patent
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Abstract
The invention provides isolated nucleic acid molecules which encode novel fatty acid desaturases and elongases from the organism Emiliana huxleyi. The invention also provides recombinant expression vectors containing desaturase or elongase nucleic acid molecules, host cells into which the expression vectors have been introduced, and methods for large-scale production of long chain polyunsaturated fatty acids (LCPUFAs), e.g. arachidonic acid (ARA), eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA).
Claims23
A polynucleotide comprising an expression control sequence operatively linked to a first nucleic acid sequence selected from the group consisting of:
- a) the nucleic acid sequence of SEQ ID NO:
- 3;
- b) a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 4;
- c) a nucleic acid sequence having at least 70% sequence identity to the nucleic acid sequence of a), wherein said nucleic acid sequence encodes a polypeptide having elongase activity;
- d) a nucleic acid sequence encoding a polypeptide having elongase activity and comprising an amino acid sequence which has at least 82% sequence identity to the amino acid sequence of SEQ ID NO:
- 4;
- and
- e) a nucleic acid sequence which is capable of hybridizing under stringent conditions comprising hybridization in 6× sodium chloride/sodium citrate (SSC) at approximately 45° C. followed by one or more wash steps in 0.2× SSC, 0.1% SDS at 50 to 65° C. to the nucleic acid sequence of a), wherein said nucleic acid sequence encodes a polypeptide having elongase activity,
- wherein said expression control sequence is heterologous to said first nucleic acid sequence,
- and wherein said polynucleotide further comprises a second nucleic acid sequence selected from the group consisting of:
- i) the nucleic acid sequence of SEQ ID NO:
- 9;
- ii) a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 10;
- iii) a nucleic acid sequence having at least 70% sequence identity to the nucleic acid sequence of i), wherein said nucleic acid sequence encodes a polypeptide having desaturase activity;
- iv) a nucleic acid sequence encoding a polypeptide having desaturase activity and comprising an amino acid sequence which has at least 82% sequence identity to the amino acid sequence of SEQ ID NO:
- 10;
- and
- v) a nucleic acid sequence which is capable of hybridizing under stringent conditions comprising hybridization in 6× sodium chloride/sodium citrate (SSC) at approximately 45° C. followed by one or more wash steps in 0.2× SSC, 0.1% SDS at 50 to 65° C. to the nucleic acid sequence of i), wherein said nucleic acid sequence encodes a polypeptide having desaturase activity.
17 more elements
A method for the manufacture of polyunsaturated fatty acids, comprising:
- a) cultivating a plant comprising a first nucleic acid sequence encoding a polypeptide having elongase activity and a second nucleic acid sequence encoding a polypeptide having desaturase activity under conditions which allow for the production of polyunsaturated fatty acids in said plant or seeds thereof;
- and
- b) obtaining said polyunsaturated fatty acids from said plant or seeds thereof,
- wherein said first nucleic acid sequence is selected from the group consisting of:
- i) the nucleic acid sequence of SEQ ID NO:
- 3;
- ii) a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 4;
- iii) a nucleic acid sequence having at least 70% sequence identity to the nucleic acid sequence of i);
- iv) a nucleic acid sequence encoding a polypeptide having at least 82% sequence identity to the amino acid sequence of SEQ ID NO:
- 4;
- and
- v) a nucleic acid sequence which is capable of hybridizing under stringent conditions comprising hybridization in 6× sodium chloride/sodium citrate (SSC) at approximately 45° C. followed by one or more wash steps in 0.2× SSC, 0.1% SDS at 50 to 65° C. to the nucleic acid sequence of i),
- and wherein said second nucleic acid sequence is selected from the group consisting of:
- i) the nucleic acid sequence of SEQ ID NO:
- 9;
- ii) a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 10;
- iii) a nucleic acid sequence having at least 70% sequence identity to the nucleic acid sequence of i);
- iv) a nucleic acid sequence encoding a polypeptide having at least 82% sequence identity to the amino acid sequence of SEQ ID NO:
- 10;
- and
- v) a nucleic acid sequence which is capable of hybridizing under stringent conditions comprising hybridization in 6× sodium chloride/sodium citrate (SSC) at approximately 45° C. followed by one or more wash steps in 0.2× SSC, 0.1% SDS at 50 to 65° C. to the nucleic acid sequence of i).
20 more elements
A method for the manufacture of an oil-, lipid- or fatty acid-composition, comprising:
- a) cultivating a plant comprising a first nucleic acid sequence encoding a polypeptide having elongase activity and a second nucleic acid sequence encoding a polypeptide having desaturase activity under conditions which allow for the production of polyunsaturated fatty acids in said plant or seeds thereof;
- and
- b) obtaining an oil-, lipid- or fatty acid-composition from said plant or seeds thereof,
- wherein said first nucleic acid sequence is selected from the group consisting of:
- i) the nucleic acid sequence of SEQ ID NO:
- 3;
- ii) a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 4;
- iii) a nucleic acid sequence having at least 70% sequence identity to the nucleic acid sequence of i);
- iv) a nucleic acid sequence encoding a polypeptide having at least 82% sequence identity to the amino acid sequence of SEQ ID NO:
- 4;
- and
- v) a nucleic acid sequence which is capable of hybridizing under stringent conditions comprising hybridization in 6× sodium chloride/sodium citrate (SSC) at approximately 45° C. followed by one or more wash steps in 0.2× SSC, 0.1% SDS at 50 to 65° C. to the nucleic acid sequence of i),
- and wherein said second nucleic acid sequence is selected from the group consisting of:
- i) the nucleic acid sequence of SEQ ID NO:
- 9;
- ii) a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 10;
- iii) a nucleic acid sequence having at least 70% sequence identity to the nucleic acid sequence of i);
- iv) a nucleic acid sequence encoding a polypeptide having at least 82% sequence identity to the amino acid sequence of SEQ ID NO:
- 10;
- and
- v) a nucleic acid sequence which is capable of hybridizing under stringent conditions comprising hybridization in 6× sodium chloride/sodium citrate (SSC) at approximately 45° C. followed by one or more wash steps in 0.2× SSC, 0.1% SDS at 50 to 65° C. to the nucleic acid sequence of i).
20 more elements
Dependencies are inferred from the claim text.