Elongase genes and uses thereof

• US 20030177508A1
• Filed: 04/04/2003
• Published: 09/18/2003
• Est. Priority Date: 08/23/1999
• Status: Active Grant

• Current Assignee: Amanda E. Leonard, Jennifer M. Thurmond, Jennifer Parker-Barnes, Pradip Mukerji, Tapas Das, Yung-Sheng Huang
• Original Assignee: Amanda E. Leonard, Jennifer M. Thurmond, Jennifer Parker-Barnes, Pradip Mukerji, Tapas Das, Yung-Sheng Huang
• Inventors: Das, Tapas, Mukerji, Pradip, Huang, Yung-Sheng, Thurmond, Jennifer M., Leonard, Amanda E., Parker-Barnes, Jennifer
• Granted Patent: US 7,070,970 B2
• Time in Patent Office: Days
• US Class Current: 800/8
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: A23L 33/12 Fatty acids or derivatives thereof
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• 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 7/6432 Eicosapentaenoic acids [EPA]
• CPC Class Codes: C12P 7/6434 Docosahexenoic acids [DHA]

Abstract

The subject invention relates to the identification of four genes involved in the elongation of polyunsaturated acids (i.e., “elongases”) and to uses thereof. Two of these genes are also involved in the elongation of monounsaturated fatty acids. In particular, elongase is utilized in the conversion of gamma linolenic acid (GLA) to dihomogamma linolenic acid (DGLA) and in the conversion of DGLA or 20:4n-3 to eicosapentaenoic acid (EPA). DGLA may be utilized in the production of polyunsaturated fatty acids, such as arachidonic acid (AA), docosahexaenoic acid (DHA), EPA, adrenic acid, ω6-docosapentaenoic acid or ω3-docosapentaenoic acid which may be added to pharmaceutical compositions, nutritional compositions, animal feeds, as well as other products such as cosmetics.

Claims

1. An isolated nucleotide sequence corresponding to or complementary to at least about 50% of the nucleotide sequence shown in SEQ ID NO:

1 (FIG.

     6).

2. The isolated nucleotide sequence of claim 1 wherein said sequence is represented by SEQ ID NO:

1.

3. The isolated nucleotide sequence of claims 1 or 2 wherein said sequence encodes a functionally active elongase which utilizes a polyunsaturated fatty acid or a monounsaturated fatty acid as a substrate.

4. The nucleotide sequence of claim 1 wherein said sequence is derived from a fungus of the genus Mortierella.

5. The nucleotide sequence of claim 4 wherein said fungus is of the species alpina.

6. A purified protein encoded by said nucleotide sequence of claims 1 or 2.

7. A purified polypeptide which elongates polyunsaturated fatty acids or monounsaturated fatty acids and has at least about 50% amino acid similarity to the amino acid sequence of said purified protein of claim 6.

45. A pharmaceutical composition comprising 1) at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 30, said another polyunsaturated fatty acid produced according to the method of claim 32, and said final polyunsaturated fatty acid produced according to the method of claim 34 and 2) a pharmaceutically acceptable carrier.

46. The pharmaceutical composition of claim 45 wherein said pharmaceutical composition is administered to a human or an animal.

47. The pharmaceutical composition of claim 45 wherein said pharmaceutical composition further comprises at least one element selected from the group consisting of a vitamin, a mineral, a salt, a carbohydrate, an amino acid, a free fatty acid, a preservative, an excipient, an anti-histamine, a growth factor, an antibiotic, a diluent, a phospholipid, an antioxidant, and a phenolic compound.

98. A pharmaceutical composition comprising 1) at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 83, said another polyunsaturated fatty acid produced according to the method of claim 85, and said final polyunsaturated fatty acid produced according to the method of claim 87 and 2) a pharmaceutically acceptable carrier.

99. The pharmaceutical composition of claim 98 wherein said pharmaceutical composition is administered to a human or an animal.

100. The pharmaceutical composition of claim 98 wherein said pharmaceutical composition further comprises an element selected from the group consisting of a vitamin, a mineral, a carbohydrate, an amino acid, a free fatty acid, a preservative, an excipient, an anti-histamine, a growth factor, an antibiotic, a phospholipid, an antioxidant, and a phenolic compound.

8. A method of producing an elongase-enzyme comprising the steps of:

a) isolating said nucleotide sequence represented by SEQ ID NO;

1 (FIG.

     6);

b) constructing a vector comprising;

i) said isolated nucleotide sequence operably linked to ii) a promoter;

c) introducing said vector into a host cell under time and conditions sufficient for expression of said elongase enzyme.

9. The method of claim 8 wherein said host cell is selected from the group consisting of a eukaryotic cell or a prokaryotic cell.

10. The method of claim 9 wherein said prokaryotic cell is selected from the group consisting of E. coli, cyanobacteria, and B. subtilis.

11. The method of claim 9 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

12. The method of claim 11 wherein said fungal cell is selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

13. The method of claim 11 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

14. The method of claim 13 wherein said yeast cell is Saccharomyces cerevisiae.

15. A vector comprising:

a) a nucleotide sequence as represented by SEQ ID NO;

1 (FIG.

     6) operably linked to b) a promoter.

16. A host cell comprising said vector of claim 15.

17. The host cell of claim 16, wherein said host cell is selected from the group consisting of a eukaryotic cell or a prokaryotic cell.

18. The host cell of claim 17 wherein said prokaryotic cell is selected from the group consisting of E. coli, Cyanobacteria, and B. subtilis.

19. The host cell of claim 17 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

20. The host cell of claim 19 wherein said fungal cell is a selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

21. The host cell of claim 20 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

22. The host cell of claim 21 wherein said host cell is Saccharomyces cerevisiae.

23. A plant cell, plant or plant tissue comprising said vector of claim 15, wherein expression of said nucleotide sequence of said vector results in production of at least one fatty acid selected from the group consisting of a monounsaturated fatty acid and a polyunsaturated fatty acid by said plant cell, plant or plant tissue.

24. The plant cell, plant or plant tissue of claim 23 wherein said polyunsaturated fatty acid is selected from the group consisting dihomo-γ

-linolenic acid (DGLA), 20;

4n-3, and adrenic acid (ADA).

25. One or more plant oils or fatty acids expressed by said plant cell, plant or plant tissue of claim 23.

26. A transgenic plant comprising said vector of claim 15, wherein expression of said nucleotide sequence of said vector results in production of a polyunsaturated fatty acid in seeds of said transgenic plant.

122. A host cell comprising said vector of claim 21.

123. The host cell of claim 122, wherein said host cell is selected from the group consisting of a eukaryotic cell or a prokaryotic cell.

124. The host cell of claim 123 wherein said prokaryotic cell is selected from the group consisting of E. coli, Cyanobacteria, and B. subtilis.

125. The host cell of claim 123 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

126. The host cell of claim 125 wherein said fungal cell is a selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

127. The host cell of claim 126 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

128. The host cell of claim 127 wherein said host cell is Saccharomyces cerevisiae.

27. A transgenic, non-human mammal whose genome comprises a DNA sequence encoding an elongase operably linked to a promoter.

28. The transgenic, non-human mammal of claim 27, wherein said DNA sequence is represented by SEQ ID NO:

1 (FIG.

     6).

29. A fluid produced by said transgenic, non-human mammal of claim 28 wherein said fluid comprises a detectable level of at least one elongase or products thereof.

30. A method for producing a polyunsaturated fatty acid comprising the steps of:

a) isolating said nucleotide sequence represented by SEQ ID NO;

1 (FIG.

     6);

b) constructing a vector comprising said isolated nucleotide sequence;

c) introducing said vector into a host cell under time and conditions sufficient for expression of elongase enzyme encoded by said isolated nucleotide sequence; and

d) exposing said expressed elongase enzyme to a substrate polyunsaturated fatty acid in order to convert said substrate to a product polyunsaturated fatty acid.

31. The method according to claim 30, wherein said substrate polyunsaturated fatty acid is selected from the group consisting of -linolenic acid (GLA), stearidonic acid (STA) and arachidonic acid (AA), and said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA, respectively.

32. The method according to claim 30 further comprising the step of exposing said product polyunsaturated fatty acid to at least one desaturase in order to convert said product polyunsaturated fatty acid to another polyunsaturated fatty acid.

33. The method according to claim 32 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA said another polyunsaturated fatty acid is selected from the group consisting of AA, eicosapentaenoic acid (EPA), ω

6-docosapentaenoic acid, respectively, and said at least one desaturase is Δ

5-desaturase, with respect to production of AA or EPA, and Δ

4-desaturase, with respect to production of ω

6-docosapentaenoic acid.

34. The method of claim 32 further comprising the step of exposing said another polyunsaturated fatty acid to one or more enzymes selected from the group consisting of at least one elongase and at least one additional desaturase in order to convert said another polyunsaturated fatty acid to a final polyunsaturated fatty acid.

35. The method of claim 34 wherein said final polyunsaturated fatty acid is selected from the group consisting of docosahexaenoic acid (DHA), AA, ω

6-docosapentaenoic acid, and ω

3-docosapentaenoic acid.

36. A nutritional composition comprising at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 30, said another polyunsaturated fatty acid produced according to the method of claim 32, and said final polyunsaturated fatty acid produced according to the method of claim 34

37. The nutritional composition of claim 36 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3 and ADA.

38. The nutritional composition of claim 36 wherein said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA, and ω

6-docosapentaenoic acid.

39. The nutritional composition of claim 36 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, adrenic acid, ω

6-docosapentaenoic acid and ω

3-docosapentaenoic acid.

40. The nutritional composition of claim 36 wherein said nutritional composition is selected from the group consisting of an infant formula, a dietary supplement and a dietary substitute.

41. The nutritional composition of claim 40 wherein said nutritional composition is administered to a human or an animal.

42. The nutritional composition of claim 41 wherein said nutritional composition is administered enterally or parenterally.

43. The nutritional composition of claim 40 wherein said nutritional composition further comprises at least one macronutrient selected from the group consisting of coconut oil, soy oil, canola oil, monoglycerides, diglycerides, triglycerides, glucose, edible lactose, electrodialysed whey, electrodialysed skim milk, milk whey, soy protein, protein hydrolysates, sunflower oil, safflower oil, corn oil, and flax oil.

44. The nutritional composition of claim 43 wherein said nutritional composition further comprises at least one vitamin selected from the group consisting of Vitamins A, C, D, E, and B complex and at least one mineral selected from the group consisting of calcium magnesium, zinc, manganese, sodium, potassium, phosphorus, copper, chloride, iodine, selenium and iron.

48. An animal feed comprising at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 30, said another polyunsaturated fatty acid produced according to the method of claim 32 and said final polyunsaturated fatty acid produced according to the method of claim 34.

49. The animal feed of claim 48 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA.

50. The animal feed of claim 48 wherein said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA, and ω

6-docosapentaenoic acid.

51. The animal feed of claim 48 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, ADA, ω

6-docosapentaenoic acid and ω

3-docosapentaenoic acid.

52. A cosmetic comprising a polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 30, said another polyunsaturated fatty acid produced according to the method of claim 32 and said final polyunsaturated fatty acid produced according to the method of claim 34.

53. A method of preventing or treating a condition caused by insufficient intake of polyunsaturated fatty acids comprising administering to said patient said nutritional composition of claim 36 in an amount sufficient to effect said prevention or treatment.

54. An isolated nucleotide sequence corresponding to or complementary to at least about 35% of the nucleotide sequence shown in SEQ ID NO:

2 (FIG.

     22).

55. The isolated nucleotide sequence of claim 54 wherein said sequence is represented by SEQ ID NO:

2.

56. The isolated nucleotide sequence of claims 54 or 55 wherein said sequence encodes a functionally active elongase which utilizes a polyunsaturated fatty acid as a substrate.

57. The nucleotide sequence of claim 55 wherein said sequence is derived from a fungus of the genus Mortierella.

58. The nucleotide sequence of claim 57 wherein said fungus is of the species alpina.

59. A purified protein encoded by said nucleotide sequence of claims 54 or 55.

60. A purified polypeptide which elongates polyunsaturated fatty acids and has at least about 30% amino acid similarity to the amino acid sequence of said purified protein of claim 59.

61. A method of producing an elongase enzyme comprising the steps of:

a) isolating said nucleotide sequence represented by SEQ ID NO;

2 (FIG.

     22);

b) constructing a vector comprising;

i) said isolated nucleotide sequence operably linked to ii) a promoter;

c) introducing said vector into a host cell under time and conditions sufficient for expression of said elongase enzyme.

62. The method of claim 61 wherein said host cell is selected from the group consisting of a eukaryotic cell or a prokaryotic cell.

63. The method of claim 62 wherein said prokaryotic cell is selected from the group consisting of E. coli, cyanobacteria, and B. subtilis.

64. The method of claim 62 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

65. The method of claim 64 wherein said fungal cell is selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

66. The method of claim 65 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

67. The method of claim 66 wherein said yeast cell is Saccharomyces cerevisiae.

68. A vector comprising:

a) a nucleotide sequence as represented by SEQ ID NO;

2 (FIG.

     22) operably linked to b) a promoter.

69. A host cell comprising said vector of claim 68.

70. The host cell of claim 69, wherein said host cell is selected from the group consisting of a eukaryotic cell or a prokaryotic cell.

71. The host cell of claim 70 wherein said prokaryotic cell is selected from the group consisting of E. coli, Cyanobacteria, and B. subtilis.

72. The host cell of claim 70 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

73. The host cell of claim 72 wherein said fungal cell is selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

74. The host cell of claim 73 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

75. The host cell of claim 74 wherein said host cell is Saccharomyces cerevisiae.

76. A plant cell, plant or plant tissue comprising said vector of claim 68, wherein expression of said nucleotide sequence of said vector results in production of a polyunsaturated fatty acid by said plant cell, plant or plant tissue.

77. The plant cell, plant or plant tissue of claim 76 wherein said polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA.

78. One or more plant oils or acids expressed by said plant cell, plant or plant tissue of claim 76.

79. A transgenic plant comprising said vector of claim 68, wherein expression of said nucleotide sequence of said vector results in production of a polyunsaturated fatty acid in seeds of said transgenic plant.

80. A transgenic, non-human mammal whose genome comprises a DNA sequence, from M. alpina, encoding an elongase operably linked to a promoter.

81. The transgenic, non-human mammal of claim 80, wherein said DNA sequence is represented by SEQ ID NO:

2 (FIG.

     22).

82. A fluid produced by said transgenic, non-human mammal of claim 81 wherein said fluid comprises a detectable level of at least one elongase or products thereof.

83. A method for producing a polyunsaturated fatty acid comprising the steps of:

a) isolating said nucleotide sequence represented by SEQ ID NO;

2 (FIG.

     22);

b) constructing a vector comprising said isolated nucleotide sequence;

c) introducing said vector into a host cell under time and conditions sufficient for expression of an elongase enzyme encoded by said isolated nucleotide sequence; and

d) exposing said expressed elongase enzyme to a substrate polyunsaturated fatty acid in order to convert said substrate to a product polyunsaturated fatty acid.

84. The method according to claim 83, wherein said substrate polyunsaturated fatty acid is selected from the group consisting of GLA, STA, and AA, and said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ω

6-docosapentaenoic acid, respectively.

85. The method according to claim 83 further comprising the step of exposing said expressed elongase enzyme to at least one desaturase in order to convert said product polyunsaturated fatty acid to another polyunsaturated fatty acid.

86. The method according to claim 85 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA, and ω

6-docosapentaenoic acid, respectively, and said at least one desaturase is Δ

5-desaturase with respect to production of AA or EPA, and Δ

4-desaturase with respect to production of ω

6-docosapentaenoic acid.

87. The method of claim 86 further comprising the steps of exposing said another polyunsaturated fatty acid to one or more enzymes selected from the group consisting of at least one elongase and at least one additional desaturase in order to convert said another polyunsaturated fatty acid to a final polyunsaturated fatty acid.

88. The method of claim 87 wherein said final polyunsaturated fatty acid is selected from the group consisting of docosahexaenoic acid, AA, ω

6-docosapentaenoic acid, and ω

3-docosapentaenoic acid.

89. A nutritional composition comprising at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 83, said another polyunsaturated fatty acid produced according to the method of claim 85, and said final polyunsaturated fatty acid produced according to the method of claim 87.

90. The nutritional composition of claim 89 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3 and ADA.

91. The nutritional composition of claim 89 wherein said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA, and ω

6-docosapentaenoic acid.

92. The nutritional composition of claim 89 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, AA, ω

6-docosapentaenoic acid, and ω

3-docosapentaenoic acid.

93. The nutritional composition of claim 89 wherein said nutritional composition is selected from the group consisting of an infant formula, a dietary supplement and a dietary substitute.

94. The nutritional composition of claim 93 wherein said nutritional composition is administered to a human or an animal.

95. The nutritional composition of claim 94 wherein said nutritional composition is administered enterally or parenterally.

96. The nutritional composition of claim 93 wherein said nutritional composition further comprises at least one macronutrient selected from the group consisting of coconut oil, soy oil, canola oil, monoglycerides, diglycerides, triglycerides, glucose, edible lactose, electrodialysed whey, electrodialysed skim milk, milk whey, soy protein, protein hydrolysates, sunflower oil, safflower oil, corn oil, and flax oil.

97. The nutritional composition of claim 96 wherein said nutritional composition further comprises at least one vitamin selected from the group consisting of Vitamins A, C, D, E, and B complex and at least one mineral selected from the group consisting of calcium magnesium, zinc, manganese, sodium, potassium, phosphorus, copper, chloride, iodine, selenium and iron.

101. An animal feed comprising at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 83, said another polyunsaturated fatty acid produced according to the method of claim 85 and said final polyunsaturated fatty acid produced according to the method of claim 87.

102. The animal feed of claim 101 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA.

103. The animal feed of claim 101 wherein said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA, and ω

6-docosapentaenoic acid.

104. The animal feed of claim 101 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, adrenic acid, ω

6-docosapentaenoic acid and ω

3-docosapentaenoic acid.

105. A cosmetic comprising a polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 83, said another polyunsaturated fatty acid produced according to the method of claim 85 and said final polyunsaturated fatty acid produced according to the method of claim 87.

106. A method of preventing or treating a condition caused by insufficient intake of polyunsaturated fatty acids comprising administering to said patient said nutritional composition of claim 89 in an amount sufficient to effect said treatment.

107. An isolated nucleotide sequence corresponding to or complementary to at least about 35% of the nucleotide sequence shown in SEQ ID NO:

3 (FIG.

     43).

108. The isolated nucleotide sequence of claim 107 wherein said sequence is represented by SEQ ID NO:

3.

109. The isolated nucleotide sequence of claims 107 or 108 wherein said sequence encodes a functionally active elongase which utilizes a polyunsaturated fatty acid or a monounsaturated fatty acid as a substrate.

110. The nucleotide sequence of claim 109 wherein said sequence is derived from a mammal.

111. The nucleotide sequence of claim 110 wherein said mammal is a human.

112. A purified protein encoded by said nucleotide sequence of claims 107 or 108.

113. A purified polypeptide which elongates polyunsaturated fatty acids or monounsaturated fatty acids and has at least about 30% amino acid similarity to the amino acid sequence of said purified protein of claim 112.

114. A method of producing an elongase enzyme comprising the steps of:

a) isolating said nucleotide sequence represented by SEQ ID NO;

3 (FIG.

     43);

b) constructing a vector comprising;

i) said isolated nucleotide sequence operably linked to ii) a promoter;

c) introducing said vector into a host cell under time and conditions sufficient for expression of said elongase enzyme.

115. The method of claim 114 wherein said host cell is selected from the group consisting of a eukaryotic cell or a prokaryotic cell.

116. The method of claim 115 wherein said prokaryotic cell is selected from the group consisting of E. coli, cyanobacteria, and B. subtilis.

117. The method of claim 115 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

118. The method of claim 117 wherein said fungal cell is selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

119. The method of claim 118 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

120. The method of claim 119 wherein said yeast cell is Saccharomyces cerevisiae.

121. A vector comprising:

a) a nucleotide sequence as represented by SEQ ID NO;

3 (FIG.

     43) operably linked to b) a promoter.

129. A plant cell, plant or plant tissue comprising said vector of claim 121, wherein expression of said nucleotide sequence of said vector results in production of at least one fatty acid selected from the group consisting of a monounsaturated fatty acid and a polyunsaturated fatty acid by said plant cell, plant or plant tissue.

130. The plant cell, plant or plant tissue of claim 129 wherein said polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3 and ADA.

131. One or more plant oils or acids expressed by said plant cell, plant or plant tissue of claim 130.

132. A transgenic plant comprising said vector of claim 121, wherein expression of said nucleotide '"'"'sequence of said vector results in production of a polyunsaturated fatty acid in seeds of said transgenic plant.

133. A transgenic, non-human mammal whose genome comprises a human DNA sequence encoding an elongase operably linked to a promoter.

134. The transgenic, non-human mammal of claim 133, wherein said DNA sequence is represented by SEQ ID NO:

3 (FIG.

     43).

135. A fluid produced by said transgenic, non-human mammal of claim 134 wherein said fluid comprises a detectable level of at least one elongase or products thereof.

136. A method for producing a polyunsaturated fatty acid comprising the steps of:

a) isolating said nucleotide sequence represented by SEQ ID NO;

3 (FIG.

     43);

b) constructing a vector comprising said isolated nucleotide sequence;

c) introducing said vector into a host cell under time and conditions sufficient for expression of elongase enzyme encoded by said isolated nucleotide sequence; and

d) exposing said expressed elongase enzyme to a substrate polyunsaturated fatty acid in order to convert said substrate to a product polyunsaturated fatty acid.

137. The method according to claim 136, wherein said substrate polyunsaturated fatty acid is selected from the group consisting of GLA, STA and AA, and said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA, respectively.

138. The method according to claim 136 further comprising the step of exposing said product polyunsaturated fatty acid to at least one desaturase in order to convert said product polyunsaturated fatty acid to another polyunsaturated fatty acid.

139. The method according to claim 138 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3 and ADA, said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA, and ω

6-docosapentaenoic acid, respectively, and said at least one desaturase is Δ

5-desaturase with respect to production of AA or EPA and Δ

4-desaturase with respect to production of ω

6-docosapentaenoic acid.

140. The method of claim 138 further comprising the step of exposing said another polyunsaturated fatty acid to one or more enzymes selected from the group consisting of at least one elongase and at least one additional desaturase in order to convert said another polyunsaturated fatty acid to a final polyunsaturated fatty acid.

141. The method of claim 140 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, ADA, ω

6-docosapentaenoic acid, and ω

3-docosapentaenoic acid.

142. A nutritional composition comprising at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 136, said another polyunsaturated fatty acid produced according to the method of claim 138, and said final polyunsaturated fatty acid produced according to the method of claim 140.

143. The nutritional composition of claim 142 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA.

144. The nutritional composition of claim 142 wherein said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA, and ω

6-docosapentaenoic acid.

145. The nutritional composition of claim 142 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, ADA, ω

6-docosapentaenoic acid, and ω

3-docosapentaenoic acid.

146. The nutritional composition of claim 142 wherein said nutritional composition is selected from the group consisting of an infant formula, a dietary supplement and a dietary substitute.

147. The nutritional composition of claim 146 wherein said nutritional composition is administered to a human or an animal.

148. The nutritional composition of claim 147 wherein said nutritional composition is administered enterally or parenterally.

149. The nutritional composition of claim 147 wherein said nutritional composition further comprises at least one macronutrient selected from the group consisting of coconut oil, soy oil, canola oil, monoglycerides, diglycerides, triglycerides, glucose, edible lactose, electrodialysed whey, electrodialysed skim milk, milk whey, soy protein, protein hydrolysates, sunflower oil, safflower oil, corn oil, and flax oil.

150. The nutritional composition of claim 149 wherein said nutritional composition further comprises at least one vitamin selected from the group consisting of Vitamins A, C, D, E, and B complex and at least one mineral selected from the group consisting of calcium magnesium, zinc, manganese, sodium, potassium, phosphorus, copper, chloride, iodine, selenium and iron.

151. A pharmaceutical composition comprising 1) at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 136, said another polyunsaturated fatty acid produced according to the method of claim 138, and said final polyunsaturated fatty acid produced according to the method of claim 140 and 2) a pharmaceutically acceptable carrier.

152. The pharmaceutical composition of claim 151 wherein said pharmaceutical composition is administered to a human or an animal.

153. The pharmaceutical composition of claim 151 wherein said pharmaceutical composition further comprises at least one element selected from the group consisting of a vitamin, a mineral, a salt, a carbohydrate, an amino acid, a free fatty acid, a preservative, an excipient, an anti-histamine, a growth factor, an antibiotic, a diluent, a phospholipid, an antioxidant, and a phenolic compound.

154. An animal feed comprising at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 136, said another polyunsaturated fatty acid produced according to the method of claim 138 and said final polyunsaturated fatty acid produced according to the method of claim 140.

155. The animal feed of claim 154 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA.

156. The animal feed of claim 154 wherein said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA, and ω

6-docosapentaenoic acid.

157. The animal feed of claim 154 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, ADA, ω

6-docosapentaenoic acid and ω

3-docosapentaenoic acid.

158. A cosmetic comprising a polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 136, said another polyunsaturated fatty acid produced according to the method of claim 138 and said final polyunsaturated fatty acid produced according to the method of claim 140.

159. A method of preventing or treating a condition caused by insufficient intake of polyunsaturated fatty acids comprising administering to said patient said nutritional composition of claim 142 in an amount sufficient to effect said treatment.

160. An isolated nucleotide sequence corresponding to or complementary to at least about 35% of the nucleotide sequence shown in SEQ ID NO:

4 (FIG.

     46).

161. The isolated nucleotide sequence of claim 160 wherein said sequence is represented by SEQ ID NO:

4.

162. The isolated nucleotide sequence of claims 160 or 161 wherein said sequence encodes a functionally active elongase which utilizes a polyunsaturated fatty acid as a substrate.

163. The nucleotide sequence of claim 162 wherein said sequence is derived from a nematode of the genus Caenorhabditis.

164. The nucleotide sequence of claim 163 wherein said nematode is of the species elegans.

165. A purified protein encoded by said nucleotide sequence of claims 160 or 161.

166. A purified polypeptide which elongates polyunsaturated fatty acids and has at least about 30% amino acid similarity to the amino acid sequence of said purified protein of claim 165.

167. A method of producing an elongase enzyme comprising the steps of:

a) isolating said nucleotide sequence represented by SEQ ID NO;

4 (FIG.

     46);

b) constructing a vector comprising;

i) said isolated nucleotide sequence operably linked to ii) a promoter;

c) introducing said vector into a host cell under time and conditions sufficient for expression of said elongase enzyme.

168. The method of claim 167 wherein said host cell is selected from the group consisting of a eukaryotic cell or a prokaryotic cell.

169. The method of claim 168 wherein said prokaryotic cell is selected from the group consisting of E. coli, cyanobacteria, and B. subtilis.

170. The method of claim 168 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

171. The method of claim 170 wherein said fungal cell is selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

172. The method of claim 171 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

173. The method of claim 172 wherein said yeast cell is Saccharomyces cerevisiae.

174. A vector comprising:

a) a nucleotide sequence as represented by SEQ ID NO;

4 (FIG.

     46) operably linked to b) a promoter.

175. A host cell comprising said vector of claim 174.

176. The host cell of claim 175, wherein said host cell is selected from the group consisting of a eukaryotic cell-or a prokaryotic cell.

177. The host cell of claim 176 wherein said prokaryotic cell is selected from the group consisting of E. coli, Cyanobacteria, and B. subtilis.

178. The host cell of claim 176 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

179. The host cell of claim 178 wherein said fungal cell is selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

180. The host, cell of claim 179 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

181. The host cell of claim 180 wherein said host cell is Saccharomyces cerevisiae.

182. A plant cell, plant or plant tissue comprising said vector of claim 174, wherein expression of said nucleotide sequence of said vector results in production of a polyunsaturated fatty acid by said plant cell, plant or plant tissue.

183. The plant cell, plant or plant tissue of claim 182 wherein said polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA.

184. One or more plant oils or acids expressed by said plant cell, plant or plant tissue of claim 182.

185. A transgenic plant comprising said vector of claim 174, wherein expression of said nucleotide sequence of said vector results in production of a polyunsaturated fatty acid in seeds of said transgenic plant.

186. A transgenic, non-human mammal whose genome comprises a DNA sequence, from C. elegans, encoding an elongase operably linked to a promoter.

187. The transgenic, non-human mammal of claim 186, wherein said DNA sequence is represented by SEQ ID NO:

4 (FIG.

     46).

188. A fluid produced by said transgenic, non-human mammal of claim 187 wherein said fluid comprises a detectable level of at least one elongase or products thereof.

189. A method for producing a polyunsaturated fatty acid comprising the steps of:

a) isolating said nucleotide sequence represented by SEQ ID NO;

4 (FIG.

     46);

b) constructing a vector comprising said isolated nucleotide sequence;

c) introducing said vector into a host cell under time and conditions sufficient for expression of an elongase enzyme encoded by said isolated nucleotide sequence; and

d) exposing said expressed elongase enzyme to a substrate polyunsaturated fatty acid in order to convert said substrate to a product polyunsaturated fatty acid.

190. The method according to claim 189, wherein said substrate polyunsaturated fatty acid is selected from the group consisting of GLA, STA, and AA and said product polyunsaturated fatty acid is DGLA, 20:

4n-3, and ADA, respectively.

191. The method according to claim 189 further comprising the step of exposing said expressed elongase enzyme to at least one desaturase in order to convert said product polyunsaturated fatty acid to another polyunsaturated fatty acid.

192. The method according to claim 191 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3 and ADA, said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA and ω

6-docosapentaenoic acid, respectively, and said at least one desaturase is Δ

5-desaturase with respect to production of AA or EPA, and Δ

4-desaturase with respect to production of ω

6-docosapentaenoic acid.

193. The method of claim 192 further comprising the steps of exposing said another polyunsaturated fatty acid to one or more enzymes selected from the group consisting of at least one elongase and at least one additional desaturase in order to convert said another polyunsaturated fatty acid to a final polyunsaturated fatty acid.

194. The method of claim 193 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, ADA, ω

6-docosapentaenoic acid, and ω

3-docosapentaenoic acid.

195. A nutritional composition comprising at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 189, said another polyunsaturated fatty acid produced according to the method of claim 191, and said final polyunsaturated fatty acid produced according to the method of claim 193.

196. The nutritional composition of claim 195 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA.

197. The nutritional composition of claim 195 wherein said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA, and ω

6-docosapentaenoic acid.

198. The nutritional composition of claim 195 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, ADA, ω

6-docosapentaenoic acid, and ω

3-docosapentaenoic acid.

199. The nutritional composition of claim 195 wherein said nutritional composition is selected from the group consisting of an infant formula, a dietary supplement and a dietary substitute.

200. The nutritional composition of claim 199 wherein said nutritional composition is administered to a human or an animal.

201. The nutritional composition of claim 200 wherein said nutritional composition is administered enterally or parenterally.

202. The nutritional composition of claim 199 wherein said nutritional composition further comprises at least one macronutrient selected from the group consisting of coconut oil, soy oil, canola oil, monoglycerides, diglycerides, triglycerides, glucose, edible lactose, electrodialysed whey, electrodialysed skim milk, milk whey, soy protein, protein hydrolysates, sunflower oil, safflower oil, corn oil and flax oil.

203. The nutritional composition of claim 202 wherein said nutritional composition further comprises at least one vitamin selected from the group consisting of Vitamins A, C, D, E, and B complex and at least one mineral selected from the group consisting of calcium magnesium, zinc, manganese, sodium, potassium, phosphorus, copper, chloride, iodine, selenium and iron.

204. A pharmaceutical composition comprising 1) at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 189, said another polyunsaturated fatty acid produced according to the method of claim 191, and said final polyunsaturated fatty acid produced according to the method of claim 193 and 2) a pharmaceutically acceptable carrier.

205. The pharmaceutical composition of claim 204 wherein said pharmaceutical composition is administered to a human or an animal.

206. The pharmaceutical composition of claim 203 wherein said pharmaceutical composition further comprises an element selected from the group consisting of a vitamin, a mineral, a carbohydrate, an amino acid, a free fatty acid, a preservative, an excipient, an anti-histamine, a growth factor, an antibiotic, a phospholipid, an antioxidant, and a phenolic compound.

207. An animal feed comprising at least one polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 189, said another polyunsaturated fatty acid produced according to the method of claim 191 and said final polyunsaturated fatty acid produced according to the method of claim 193.

208. The animal feed of claim 207 wherein said product polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3 and ADA.

209. The animal feed of claim 207 wherein said another polyunsaturated fatty acid is selected from the group consisting of AA, EPA and ω

6-docosapentaenoic acid.

210. The animal feed of claim 207 wherein said final polyunsaturated fatty acid is selected from the group consisting of DHA, ADA, ω

6-docosapentaenoic acid and ω

3-docosapentaenoic acid.

211. A cosmetic comprising a polyunsaturated fatty acid selected from the group consisting of said product polyunsaturated fatty acid produced according to the method of claim 189, said another polyunsaturated fatty acid produced according to the method of claim 191 and said final polyunsaturated fatty acid produced according to the method of claim 193.

212. A method of preventing or treating a condition caused by insufficient intake of polyunsaturated fatty acids comprising administering to said patient said nutritional composition of claim 195 in an amount sufficient to effect said prevention or treatment.