Method For the Production of Multiple-Unsaturated Fatty Acids in Transgenic Organisms

• US 20080155705A1
• Filed: 07/16/2004
• Published: 06/26/2008
• Est. Priority Date: 08/01/2003
• Status: Active Grant

• Current Assignee: BASF Plant Science GmbH (BASF SE)
• Original Assignee: BASF Plant Science GmbH (BASF SE)
• Inventors: Meyer, Astrid, Sperling, Petra, Cirpus, Petra, Zank, Thorsten, Abbadi, Amine, Qiu, Xiao, Kirsch, Jelena, Heinz, Ernst, Vrinten, Patricia, Domergue, Frederic, Bauer, Jorg
• Granted Patent: US 9,433,228 B2
• Time in Patent Office: Days
• US Class Current: 800/13
• CPC Class Codes: A01H 1/00 Processes for modifying genotypes ; Plants characterised by associated natural traits A01H4/00 takes precedence
• CPC Class Codes: A23D 9/00 Other edible oils or fats, e.g. shortenings, cooking oils
• CPC Class Codes: A23D 9/02 characterised by the production or working-up
• CPC Class Codes: A23L 33/115 Fatty acids or derivatives thereof; Fats or oils
• CPC Class Codes: A23V 2002/00 Food compositions, function of food ingredients or processes for food or foodstuffs
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: C11B 1/00 Production of fats or fatty oils from raw materials
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12N 9/0083 Miscellaneous (1.14.99)
• CPC Class Codes: C12N 9/0093 acting on CH or CH2 groups (1.17)
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12P 7/6409 Fatty acids
• CPC Class Codes: C12P 7/6427 Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12P 7/6431 Linoleic acids [18:2[n-6]]
• CPC Class Codes: C12P 7/6432 Eicosapentaenoic acids [EPA]
• CPC Class Codes: C12P 7/6434 Docosahexenoic acids [DHA]
• CPC Class Codes: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12P 7/6472 containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12Y 114/19 with oxidation of a pair of donors resulting in the reduction of molecular oxygen to two molecules of water (1.14.19)
• CPC Class Codes: C12Y 114/19003 : Linoleoyl-CoA desaturase (1.14.19.3)
• CPC Class Codes: C12Y 203/01199 : Very-long-chain 3-oxoacyl-CoA synthase (2.3.1.199)
• CPC Class Codes: Y02P 20/52 : using catalysts, e.g. selective catalysts

Abstract

The invention relates to a method for the production of multiply-unsaturated fatty acids in an organism, into which nucleic acids have been introduced, which code for polypeptides with Δ-5 elongase activity. Said nucleic acid sequences, optionally with further nucleic acid sequences, coding for polypeptides for the biosynthesis of fatty acids and lipid metabolism, are advantageously expressed in the organism. Nucleic acid sequences coding for a Δ-6 desaturase, a Δ-5 desaturase, Δ-4 desaturase and/or Δ-6 elongase activity are particularly advantageous and, advantageously, said saturases and elongases are derived from Thalassiosira, Euglena or Ostreococcus. The invention further relates to a method for the production of oils and/or triacylglycerides with an increased content of long-chain, multiply-unsaturated fatty acids. A particular embodiment of the invention is a method for the production of unsaturated ω-3 fatty acids and a method for the production of triglycerides with an increased content of unsaturated fatty acids, in particular, of Δ-3 fatty acids with more than three double bonds. Also disclosed is the production of a transgenic organism, preferably a transgenic plant, or a transgenic microorganism with increased content of ω-3 fatty acids, oils or lipids with ω-3 double bonds as a result of the expression of the elongases and desaturases employed in the above method, preferably in combination with ω-3 desaturases, for example a ω-3 desaturase from fungi of the family Pythiaceae such as the genus Phytophtora, for example, the genus and species Phytophtora infestans, or a ω-3 desaturase from algae such as the family Prasinophyceae, for example, the genus Ostreococcus and, particularly, the genus and species Ostreococcus tauri or diatomaceae such as the genus Thalassiosira and, particularly, the genus and species Thalassiosira pseudonana. The invention also relates to the nucleic acid sequences, nucleic acid constructs, vectors and organisms containing the nucleic acid sequences and/or the nucleic acid constructs and transgenic organisms containing said nucleic acid sequences, nucleic acid constructs and/or vectors. A further part of the invention relates to oils, lipids and/or fatty acids produced according to the above method and use thereof and, furthermore, unsaturated fatty acids and triglycerides with an increased content of unsaturated fatty acids and use thereof.

Claims

1. A process for the production of compounds of the formula I

2. The process according to claim 1, wherein the nucleic acid sequences which encode polypeptides with Δ

9-elongase, Δ

6-desaturase, Δ

8-desaturase, Δ

6-elongase, Δ

5-desaturase, Δ

5-elongase or Δ

4-desaturase activity are selected from the group consisting of;

a) a nucleic acid sequence with the sequence shown in SEQ ID NO;

1, SEQ ID NO;

3, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

15, SEQ ID NO;

17, SEQ ID NO;

19, SEQ ID NO;

21, SEQ ID NO;

23, SEQ ID NO;

25, SEQ ID NO;

27, SEQ ID NO;

29, SEQ ID NO;

31, SEQ ID NO;

33, SEQ ID NO;

35, SEQ ID NO;

37, SEQ ID NO;

39, SEQ ID NO;

41, SEQ ID NO;

43, SEQ ID NO;

45, SEQ ID NO;

47, SEQ ID NO;

49, SEQ ID NO;

51, SEQ ID NO;

53, SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65, SEQ ID NO;

67, SEQ ID NO;

69, SEQ ID NO;

71, SEQ ID NO;

73, SEQ ID NO;

75, SEQ ID NO;

77, SEQ ID NO;

79, SEQ ID NO;

81, SEQ ID NO;

83, SEQ ID NO;

85, SEQ ID NO;

89, SEQ ID NO;

91, SEQ ID NO;

93, SEQ ID NO;

95, SEQ ID NO;

97, SEQ ID NO;

99, SEQ ID NO;

101, SEQ ID NO;

103, SEQ ID NO;

111, SEQ ID NO;

113, SEQ ID NO;

117, SEQ ID NO;

119, SEQ ID NO;

131, SEQ ID NO;

133, SEQ ID NO;

135, SEQ ID NO;

137 or SEQ ID NO;

183, or

b) nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequences shown in SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

6, SEQ ID NO;

8, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

20, SEQ ID NO;

22, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, SEQ ID NO;

38, SEQ ID NO;

40, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

46, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

52, SEQ ID NO;

54, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

70, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

76, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

82, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

88, SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

96, SEQ ID NO;

98, SEQ ID NO;

100, SEQ ID NO;

102, SEQ ID NO;

104, SEQ ID NO;

112, SEQ ID NO;

114, SEQ ID NO;

118, SEQ ID NO;

120, SEQ ID NO;

132, SEQ ID NO;

134, SEQ ID NO;

136, SEQ ID NO;

138 or SEQ ID NO;

184, or

c) derivatives of the nucleic acid sequence shown in SEQ ID NO;

1, SEQ ID NO;

3, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

15, SEQ ID NO;

17, SEQ ID NO;

19, SEQ ID NO;

21, SEQ ID NO;

23, SEQ ID NO;

25, SEQ ID NO;

27, SEQ ID NO;

29, SEQ ID NO;

31, SEQ ID NO;

33, SEQ ID NO;

35, SEQ ID NO;

37, SEQ ID NO;

39, SEQ ID NO;

41, SEQ ID NO;

43, SEQ ID NO;

45, SEQ ID NO;

47, SEQ ID NO;

49, SEQ ID NO;

51, SEQ ID NO;

53, SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65, SEQ ID NO;

67, SEQ ID NO;

69, SEQ ID NO;

71, SEQ ID NO;

73, SEQ ID NO;

75, SEQ ID NO;

77, SEQ ID NO;

79, SEQ ID NO;

81, SEQ ID NO;

83, SEQ ID NO;

85, SEQ ID NO;

89, SEQ ID NO;

91, SEQ ID NO;

93, SEQ ID NO;

95, SEQ ID NO;

97, SEQ ID NO;

99, SEQ ID NO;

101, SEQ ID NO;

103, SEQ ID NO;

11, SEQ ID NO;

113, SEQ ID NO;

117, SEQ ID NO;

119, SEQ ID NO;

131, SEQ ID NO;

133, SEQ ID NO;

135, SEQ ID NO;

137 or SEQ ID NO;

183 which encode polypeptides with at least 40% identity at the amino acid level with SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

6, SEQ ID NO;

8, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

20, SEQ ID NO;

22, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, SEQ ID NO;

38, SEQ ID NO;

40, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

46, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

52, SEQ ID NO;

54, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

70, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

76, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

82, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

88, SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

96, SEQ ID NO;

98, SEQ ID NO;

100, SEQ ID NO;

102, SEQ ID NO;

104, SEQ ID NO;

112, SEQ ID NO;

114, SEQ ID NO;

118, SEQ ID NO;

120, SEQ ID NO;

132, SEQ ID NO;

134, SEQ ID NO;

136, SEQ ID NO;

138 or SEQ ID NO;

184 and which have Δ

9-elongase, Δ

6-desaturase, Δ

8-desaturase, Δ

6-elongase, Δ

5-desaturase, Δ

5-elongase or Δ

4-desaturase activity.

3. The process according to claim 1, wherein a nucleic acid sequence which encodes polypeptides with ω

3-desaturase activity, selected from the group consisting of;

a. a nucleic acid sequence with the sequence shown in SEQ ID NO;

87 or SEQ ID NO;

105, or

b. nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO;

88 or SEQ ID NO;

106, or

c. derivatives of the nucleic acid sequence shown in SEQ ID NO;

87 or SEQ ID NO;

105 which encodes polypeptides with at least 60% identity at the amino acid level with SEQ ID NO;

88 or SEQ ID NO;

106 and which have ω

3-desaturase activity is additionally introduced to the organism.

4. The process according to claim 1, wherein a nucleic acid sequence which encodes polypeptides with Δ

12-desaturase activity, selected from the group consisting of;

a) a nucleic acid sequence with the sequence shown in SEQ ID NO;

107 or SEQ ID NO;

109, or

b) nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO;

108 or SEQ ID NO;

110, or

c) derivatives of the nucleic acid sequence shown in SEQ ID NO;

107 or SEQ ID NO;

110 which encode polypeptides with at least 60% identity at the amino acid level with SEQ ID NO;

108 or SEQ ID NO;

110 and which have Δ

12-desaturase activity is additionally introduced into the organism.

5. The process according to claim 1, wherein the substituents R² or R³ independently of one another are saturated or unsaturated C₁₈-C₂₂-alkylcarbonyl.

6. The process according to claim 1, wherein the substituents R² or R³ independently of one another are unsaturated C₁₈-, C₂₀- or C₂₂-alkyl-carbonyl with at least two double bonds.

7. The process according to claim 1, wherein the transgenic organism is a transgenic microorganism or a transgenic plant.

8. The process according to claim 1, wherein the transgenic organism is an oil-producing plant, a vegetable plant or an ornamental.

9. The process according to claim 1, wherein the transgenic organism is a transgenic plant selected from the group consisting of the plant families:

Adelotheciaceae, Anacardiaceae, Asteraceae, Apiaceae, Betulaceae, Boraginaceae, Brassicaceae, Bromeliaceae, Caricaceae, Cannabaceae, Convolvulaceae, Chenopodiaceae, Crypthecodiniaceae, Cucurbitaceae, Ditrichaceae, Elaeagnaceae, Ericaceae, Euphorbiaceae, Fabaceae, Geraniaceae, Gramineae, Juglandaceae, Lauraceae, Leguminosae, Linaceae of and Prasinophyceae.

10. The process according to claim 1, wherein the compounds of the formula I are isolated from the organism in the form of oils, lipids or free fatty acids.

11. The process according to claim 1, wherein the compounds of the formula I are isolated in a concentration of at least 5% by weight based on the total lipid content of the transgenic organism.

12. Oil, lipid or fatty acid, or a fraction thereof, produced by the process according to claim 1.

13. Oil, lipid or fatty acid composition which comprises polyunsaturated fatty acids (PUFAs) produced by the process according to claim 1 and which is derived from transgenic plants.

14. A process for the production of oils, lipids or fatty acid compositions, wherein the process comprises mixing oils, lipids or fatty acids produced by the process according to claim 1, or oil, lipid or fatty acid compositions comprising polyunsaturated fatty acids (PUFAs) produced by the process according to claim 1 and derived from transgenic plants with animal oils, lipids or fatty acids.

15. Feed, foodstuffs, cosmetics or pharmaceuticals comprising oils, lipids or fatty acids produced by the process according to claim 1, or oils, lipid or fatty acid compositions comprising polyunsaturated fatty acids (PUFAs) produced by the process according to claim 1 and derived from transgenic plants.

16. An isolated nucleic acid sequence which encodes a polypeptide with Δ

5-elongase activity and comprises an amino acid sequence selected from the group of amino acid sequences as shown in SEQ ID NO;

115, SEQ ID NO;

116, SEQ ID NO;

139, SEQ ID NO;

140, SEQ ID NO;

141 of and SEQ ID NO;

142.

17. The isolated nucleic acid sequence according to claim 16, wherein the nucleic acid sequence which encodes a polypeptide with Δ

5-elongase activity comprises a combination of the amino acid sequences selected from the group consisting of;

a) SEQ ID NO;

115 and SEQ ID NO;

139, SEQ ID NO;

115 and SEQ ID NO;

140 or SEQ ID NO;

139 and SEQ ID NO;

140;

or

b) SEQ ID NO;

116 and SEQ ID NO;

141, SEQ ID NO;

116 and SEQ ID NO;

142 or SEQ ID NO;

141 and SEQ ID NO;

142;

or

c) SEQ ID NO;

115, SEQ ID NO;

139 and SEQ ID NO;

140 or SEQ ID NO;

116, SEQ ID NO;

141 and SEQ ID NO;

142.

18. The isolated nucleic acid sequence according to claim 16 which encodes a polypeptide with Δ

5-elongase activity, wherein the nucleic acid sequence is selected from the group consisting of;

a. a nucleic acid sequence with the sequence shown in SEQ ID NO;

43, SEQ ID NO;

45, SEQ ID NO;

47, SEQ ID NO;

49, SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65, SEQ ID NO;

67, SEQ ID NO;

75, SEQ ID NO;

77, SEQ ID NO;

79, SEQ ID NO;

83, SEQ ID NO;

85, SEQ ID NO;

113, SEQ ID NO;

131 or SEQ ID NO;

133,

b. nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO;

44, SEQ ID NO;

46, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

76, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

114, SEQ ID NO;

132 or SEQ ID NO;

134, or

c. derivatives of the nucleic acid sequence shown in SEQ ID NO;

43, SEQ ID NO;

45, SEQ ID NO;

47, SEQ ID NO;

49, SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65, SEQ ID NO;

67, SEQ ID NO;

75, SEQ ID NO;

77, SEQ ID NO;

79, SEQ ID NO;

83, SEQ ID NO;

85, SEQ ID NO;

113, SEQ ID NO;

131 or SEQ ID NO;

133 which encode polypeptides with at least 40% homology at the amino acid level with SEQ ID NO;

44, SEQ ID NO;

46, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

76, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

114, SEQ ID NO;

132 or SEQ ID NO;

134 and which have Δ

5-elongase activity.

25. The isolated nucleic acid sequence according to claim 16, which sequence is derived from an alga, a fungus, a microorganism, a plant or a nonhuman animal.

26. The isolated nucleic acid sequence according to claim 16, which sequence is derived from the order Salmoniformes, the diatom genera Thalassiosira or Crypthecodinium or from the family of the Prasinophyceae, Euglenaceae or Pythiaceae.

27. An amino acid sequence which is encoded by an isolated nucleic acid sequence according to claim 16.

28. A gene construct comprising an isolated nucleic acid according to claim 16, wherein the nucleic acid is linked operably with one or more regulatory signals.

29. The gene construct according to claim 28, wherein the nucleic acid construct comprises additional biosynthesis genes of the fatty acid or lipid metabolism selected from the group acyl-CoA dehydrogenase(s), acyl-ACP [=acyl carrier protein] desaturase(s), acyl-ACP thioesterase(s), fatty acid acyltransferase(s), acyl-CoA:

lysophospholipid acyltransferase(s), fatty acid synthase(s), fatty acid hydroxylase(s), acetyl-coenzyme A carboxylase(s), acyl-coenzyme A oxidase(s), fatty acid desaturase(s), fatty acid acetylenases, lipoxygenases, triacylglycerol lipases, allenoxide synthases, hydroperoxide lyases or fatty acid elongase(s).

30. The gene construct according to claim 28, wherein the nucleic acid construct comprises additional biosynthesis genes of the fatty acid or lipid metabolism selected from the group Δ

4-desaturase, Δ

5-desaturase, Δ

6-desaturase, Δ

8-desaturase, Δ

9-desaturase, Δ

12-desaturase, Δ

6-elongase or Δ

9-elongase.

31. A vector comprising a the nucleic acid according to claim 16.

32. A transgenic nonhuman organism, at least one nucleic acid according to claim 16.

33. The transgenic nonhuman organism according to claim 32, which organism is a microorganism, a nonhuman animal or a plant.

34. The transgenic nonhuman organism according to claim 32, which organism is a plant.

19. An isolated nucleic acid sequence which encodes a polypeptide with Δ

6-elongase activity selected from the group consisting of;

a. a nucleic acid sequence with the sequence shown in SEQ ID NO;

69, SEQ ID NO;

81, SEQ ID NO;

111 or SEQ ID NO;

183,

b. nucleic acid sequences which, as a result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO;

70, SEQ ID NO;

82, SEQ ID NO;

112 or SEQ ID NO;

184, or

c. derivatives of the nucleic acid sequence shown in SEQ ID NO;

69, SEQ ID NO;

81, SEQ ID NO;

111 or SEQ ID NO;

183 which encode polypeptides with at least 40% homology at the amino acid level with SEQ ID NO;

70, SEQ ID NO;

82, SEQ ID NO;

112 or SEQ ID NO;

184 and which have Δ

6-elongase activity.

20. An isolated nucleic acid sequence which encodes a polypeptide with ω

3-desaturase activity, selected from the group consisting of;

a. a nucleic acid sequence with the sequence shown in SEQ ID NO;

87 or SEQ ID NO;

105,

b. nucleic acid sequences which, as a result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO;

88 or SEQ ID NO;

106, or

c. derivatives of the nucleic acid sequence shown in SEQ ID NO;

87 or SEQ ID NO;

105 which have polypeptides with at least 60% identity at the amino acid level with SEQ ID NO;

88 or SEQ ID NO;

106 and which have ω

3-desaturase activity.

21. An isolated nucleic acid sequence which encodes a polypeptide with Δ

6-desaturase activity, selected from the group consisting of;

a) a nucleic acid sequence with the sequence shown in SEQ ID NO;

89 or in SEQ ID NO;

97,

b) nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO;

90 or in SEQ ID NO;

98, or

c) derivatives of the nucleic acid sequence shown in SEQ ID NO;

89 or in SEQ ID NO;

97 which encode polypeptides with at least 40% homology at the amino acid level with SEQ ID NO;

90 or in SEQ ID NO;

98 and which have Δ

6-desaturase activity.

22. An isolated nucleic acid sequence which encodes a polypeptide with Δ

5-desaturase activity, selected from the group consisting of;

a) a nucleic acid sequence with the sequence shown in SEQ ID NO;

91, SEQ ID NO;

93, SEQ ID NO;

99 or in SEQ ID NO;

101,

b) nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

100, or in SEQ ID NO;

102, or

c) derivatives of the nucleic acid sequence shown in SEQ ID NO;

91, SEQ ID NO;

93, SEQ ID NO;

99 or in SEQ ID NO;

101 which encode polypeptides with at least 40% homology at the amino acid level with SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

100, or in SEQ ID NO;

102 and which have Δ

5-desaturase activity.

23. An isolated nucleic acid sequence which encodes a polypeptide with Δ

4-desaturase activity, selected from the group consisting of;

a) a nucleic acid sequence with the sequence shown in SEQ ID NO;

95 or in SEQ ID NO;

103,

b) nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO;

96 or in SEQ ID NO;

104, or

c) derivatives of the nucleic acid sequence shown in SEQ ID NO;

95 or in SEQ ID NO;

103 which encode polypeptides with at least 40% homology at the amino acid level with SEQ ID NO;

96 or in SEQ ID NO;

104 and which have Δ

4-desaturase activity.

24. An isolated nucleic acid sequence which encodes a polypeptide with Δ

12-desaturase activity, selected from the group consisting of;

a) a nucleic acid sequence with the sequence shown in SEQ ID NO;

107 or in SEQ ID NO;

109, or

b) nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO;

108 or in SEQ ID NO;

110, or

c) derivatives of the nucleic acid sequence shown in SEQ ID NO;

107 or in SEQ ID NO;

109 which encode polypeptides with at least 50% identity at the amino acid level with SEQ ID NO;

108 or in SEQ ID NO;

110 and which have Δ

12-desaturase activity.