LIPID COMPOSITIONS COMPRISING TRIACYLGLYCEROL WITH LONG-CHAIN POLYUNSATURATED FATTY ACIDS AT THE SN-2 POSITION

• US 20160177220A1
• Filed: 12/18/2015
• Published: 06/23/2016
• Est. Priority Date: 06/27/2014
• Status: Active Grant

• Current Assignee: Commonwealth Scientific & Industrial Research Organisation (Commonwealth Of Australia As Represented By Department Of Industry, Innovation, Climate Change, Science, Research And Tertiary Education), Grains Research & Development Corporation, Nuseed Nutritional Australia Pty Limited
• Original Assignee: Commonwealth Scientific & Industrial Research Organisation (Commonwealth Of Australia As Represented By Department Of Industry, Innovation, Climate Change, Science, Research And Tertiary Education), Grains Research & Development Corporation, Nuseed Pty. Ltd. (Nufarm Ltd.)
• Inventors: Petrie, James Robertson, Singh, Surinder Pal
• Granted Patent: US 10,005,713 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A23D 9/00 Other edible oils or fats, e.g. shortenings, cooking oils
• CPC Class Codes: A61K 2236/00 Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
• CPC Class Codes: A61K 31/20 having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
• CPC Class Codes: A61K 31/201 having one or two double bonds, e.g. oleic, linoleic acids
• CPC Class Codes: A61K 31/202 having three or more double bonds, e.g. linolenic eicosanoids, e.g. leukotrienes A61K31/557
• CPC Class Codes: A61K 31/23 of acids having a carboxyl group bound to a chain of seven or more carbon atoms
• CPC Class Codes: A61K 31/232 having three or more double bonds, e.g. etretinate
• CPC Class Codes: A61K 36/185 Magnoliopsida (dicotyledons)
• CPC Class Codes: A61K 36/31 Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
• CPC Class Codes: C07C 69/587 Monocarboxylic acid esters having at least two carbon-to-carbon double bonds
• CPC Class Codes: C11B 1/10 by extracting
• CPC Class Codes: C11B 3/02 by chemical reaction C11B3/003 takes precedence
• CPC Class Codes: C11B 3/04 with acids
• CPC Class Codes: C11B 3/10 by adsorption
• CPC Class Codes: C11B 3/12 by distillation
• CPC Class Codes: C11B 7/0075 by differences of melting or solidifying points
• CPC Class Codes: C11C 3/003 by esterification of fatty acids with alcohols C11C3/02 takes precedence
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition

Abstract

The present invention relates to lipid comprising docosapentaenoic acid and/or docosahexaenoic acid preferentially esterified at the sn-2 position of triacylglycerol, and processes for producing and using the lipid.

Claims

1. Extracted plant lipid comprising fatty acids in an esterified form, the fatty acids comprising palmitic acid and C22 polyunsaturated fatty acid which comprises docosapentaenoic acid (DPA) and/or docosahexaenoic acid (DHA), and optionally myristic acid, wherein at least 35% of the DPA and/or DHA esterified in the form of triacylglycerol (TAG) is esterified at the sn-2 position of the TAG, wherein the level of palmitic acid in the total fatty acid content of the extracted lipid is between about 2% and 16%, and wherein the level of myristic acid (C14:

0) in the total fatty acid content of the extracted lipid, if present, is less than 1%.

2. The lipid of claim 1 which has one or more of the following features

i) the fatty acids further comprise one or more or all of oleic acid, ω

6 fatty acids which comprise linoleic acid (LA), ω

3 fatty acids which comprise α

-linolenic acid (ALA) and optionally one or more or all of stearidonic acid (SDA), eicosapentaenoic acid (EPA), and eicosatetraenoic acid (ETA),

ii) at least about 40%, at least about 45%, at least about 48%, between 35% and about 60%, or between 35% and about 50%, of the DPA and/or DHA esterified in the form of triacylglycerol (TAG) is esterified at the sn-2 position of the TAG,

iii) the triacyiglycerol (TAG) content of the lipid is at least about 70%, at least about 80%, at least about 90%, at least 95%, between about 70% and about 99%, or between about 90% and about 99%, and

iv) the level of DPA and/or DHA in the total fatty acid content of the extracted lipid is between about 1% and 35%, or between about 7% and 35% or between about 20.1% and 35%.

3. The lipid of claim 2, wherein the level of DPA and/or DHA in the total fatty acid content of the extracted lipid is about 7%, about 8%, about 9%, about 10%, about 12%, about 15%, about 18%, about 20%, about 22%, about 24%, about 26%, about 28%, about 30%, between about 7% and about 28%, between about 7% and about 25%, between about 10% and 35%, between about 10% and about 30%, between about 10% and about 25%, between about 10% and about 22%, between about 14% and 35%, between about 16% and 35%, between about 16% and about 30%, between about 16% and about 25%, or between about 16% and about 22%.

4. The lipid of claim 1 which has one or more of the following features

i) the level of palmitic acid in the total fatty acid content of the extracted lipid is between about 2% and 15%, or between about 3% and about 10%,

ii) the level of myristic acid (C14:

0) in the total fatty acid content of the extracted lipid is about 0.1%,

iii) the level of oleic acid in the total fatty acid content of the extracted lipid is between about 1% and about 30%, between about 3% and about 30%, between about 6% and about 30%, between 1% and about 20%, between about 30% and about 60%, about 45% to about 60%, about 30%, or between about 15% and about 30%,

iv) the level of linoleic acid (LA) in the total fatty acid content of the extracted lipid is between about 4% and about 35%, between about 4% and about 20%, between about 4% and about 17%, or between about 5% and about 10%,

v) the level of α

-linolenic acid (ALA) in the total fatty acid content of the extracted lipid is between about 4% and about 40%, between about 7% and about 40%, between about 10% and about 35%, between about 20% and about 35%, between about 4% and 16%, or between about 2% and 16%,

vi) the level of γ

-linolenic acid (GLA) in the total fatty acid content of the extracted lipid is less than 4%, less than about 3%, less than about 2%, less than about 1%, less than about 0.5%, between 0.05% and 7%, between 0.05% and 4%, between 0.05% and about 3%, or between 0.05% and about 2%,

vii) the level of stearidonic acid (SDA) in the total fatty acid content of the extracted lipid is less than about 10%, less than about 8%, less than about 7%, less than about 6%, less than about 4%, less than about 3%, between about 0.05% and about 7%, between about 0.05% and about 6%, between about 0.05% and about 4%, between about 0.05% and about 3%, between about 0.05% and about 10%, or between 0.05% and about 2%,

viii) the level of eicosatetraenoic acid (ETA) in the total fatty acid content of the extracted lipid is less than about 6%, less than about 5%, less than about 4%, less than about 1%, less than about 0.5%, between 0.05% and about 6%, between 0.05% and about 5%, between 0.05% and about 4%, between 0.05% and about 3%, or between 0.05% and about 2%,

ix) the level of eicosatrienoic acid (ETrA) in the total fatty acid content of the extracted lipid is less than 4%, less than about 2%, less than about 1%, between 0.05% and 4%, between 0.05% and 3%, or between 0.05% and about 2%, or between 0.05% and about 1%,

x) the level of eicosapentaenoic acid (EPA) in the total fatty acid content of the extracted lipid is between 4% and 15%, less than 4%, less than about 3%, less than about 2%, between 0.05% and 10%, between 0.05% and 5%,

between 0.05% and about 3%, or between 0.05% and about 2%,

xi) the lipid comprises ω

6-docosapentaenoic acid (22;

5<sup>Δ

4,7,10,13,16</sup>) in its fatty acid content,

xii) the lipid comprises less than 0.1% of ω

6-docosapentaenoic acid (22;

5<sup>Δ

4,7,10,13,16</sup>) in its fatty acid content,

xiii) the lipid comprises less than 0.1% of one or more or all of SDA, EPA and ETA in its fatty acid content,

xiv) the level of total saturated fatty acids in the total fatty acid content of the extracted lipid is between about 4% and about 25%, between about 4% and about 20%, between about 6% and about 20%, or between about 6% and about 12%,

xv) the level of total monounsaturated fatty acids in the total fatty acid content of the extracted lipid is between about 4^% and about 40%, between about 4% and about 35%, between about 8% and about 25%, between 8% and about 22%, between about 15% and about 40% or between about 15% and about 35%,

xvi) the level of total polyunsaturated fatty acids in the total fatty acid content of the extracted lipid is between about 20% and about 75%, between 30% and 75%, between about 50% and about 75%, about 60%, about 65%, about 70%, about 75%, or between about 60% and about 75%,

xvii) the level of total ω

6 fatty acids in the total fatty acid content of the extracted lipid is between about 35% and about 50%, between about 20% and about 35%, between about 6% and 20%, less than 20%, less than about 16%, less than about 10%, between about 1% and about 16%, between about 2% and about 10%, or between about 4% and about 10%,

xviii) the level of new ω

6 fatty acids in the total fatty acid content of the extracted lipid is less than about 10%, less than about 8%, less than about 6%, less than 4%, between about 1% and about 20%, between about 1% and about 10%, between 0.5% and about 8%, or between 0.5% and 4%,

xix) the level of total ω

3 fatty acids in the total fatty acid content of the extracted lipid is between 36% and about 65%, between 36% and about 70%, between 40% and about 60%, between about 30% and about 60%, between about 35% and about 60%, between 40% and about 65%, between about 30% and about 65%, between about 35% and about 65%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65% or about 70%,

xx) the level of new ω

3 fatty acids in the total fatty acid content of the extracted lipid is between 21% and about 45%, between 21% and about 35%, between about 23% and about 35%, between about 25% and about 35%, between about 27% and about 35%, about 23%, about 25%, about 27%, about 30%, about 35%, about 40% or about 45%,

xxi) the ratio of total ω

6 fatty acids;

total ω

3 fatty acids in the fatty acid content of the extracted lipid is between about 1.0 and about 3.0, between about 0.1 and about 1, between about 0.1 and about 0.5, less than about 0.50, less than about 0.40, less than about 0.30, less than about 0.20, less than about 0.15, about 1.0, about 0.1, about 0.10 to about 0.4, or about 0.2,

xxii) the ratio of new ω

6 fatty acids;

new ω

3 fatty acids in the fatty acid content of the extracted lipid is between about 1.0 and about 3.0, between about 0.02 and about 0.1, between about 0.1 and about 1, between about 0.1 and about 0.5, less than about 0.50, less than about 0.40, less than about 0.30, less than about 0.20, less than about 0.15, about 0.02, about 0.05, about 0.1, about 0.2 or about 1.0,

xxiii) the fatty acid composition of the lipid is based on an efficiency of conversion of ALA to DPA and/or DHA of at least about 17%, at least about 22%, at least about 24%, at least about 30%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, between about 22% and about 70%, between about 17% and about 55%, between about 22% and about 40%, or between about 24% and about 40%,

xxiv) the total fatty acid in the extracted lipid has less than 1.5% C20;

1, less than 1% C20;

1 or about 1% C20;

1,

xxv) the lipid comprises diacylglycerol (DAG), which DAG preferably comprises DPA and/or DHA,

xxvi) the lipid comprises less than about 10%, less than about 5%, less than about 1%, or between about 0.001% and about 5%, free (non-esterified) fatty acids and/or phospholipid, or is essentially free thereof,

xxvii) the most abundant DPA and/or DHA-containing TAG species in the lipid is DPA/8;

3/18;

3 (TAG 56;

12) and/or DHA/18;

38;

3/18;

3 (TAG 56;

12), and

xxviii) the lipid comprises tri-DPA TAG (TAG 66;

18) and/or tri-DHA TAG (TAG 66;

18).

5. A process for producing extracted plant lipid, comprising the steps of

i) obtaining a plant part or plurality of plant parts comprising lipid, the lipid comprising fatty acids in an esterified form, the fatty acids comprising palmitic acid and C22 polyunsaturated fatty acid which comprises docosapentaenoic acid (DPA) and/or docosahexaenoic acid (DHA), and optionally myristic acid, wherein at least 35% of the DPA and/or DHA esterified in the form of triacylglycerol (TAG) is esterified at the sn-2 position of the TAG, wherein the level of palmitic acid in the total fatty acid content of the extracted lipid is between about 2% and 16%, and wherein the level of myristic acid (C14:

0) in the total fatty acid content of the extracted lipid, if present, is less than 1%, and

ii) extracting lipid from the plant part or plurality of plant parts.

6. The process of claim 5, wherein lipid has one or more of the following features

i) the fatty acids further comprise one or more or all of oleic acid, ω

6 fatty acids which comprise linoleic acid (LA), ω

3 fatty acids which comprise α

-linolenic acid (ALA) and optionally one or more of stearidonic acid (SDA), eicosapentaenoic acid (EPA), and eicosatetraenoic acid (ETA),

ii) at least about 40%, at least about 45%, at least about 48%, between 35% and about 60%, or between 35% and about 50%, of the DPA and/or DHA esterified in the form of triacylglycerol (TAG) is esterified at the sn-2 position of the TAG,

iii) the triacylglycerol (TAG) content of the lipid is at least about 70%, at least about 80%, at least about 90%, at least 95%, between about 70% and about 99%, or between about 90% and about 99%, and

iv) the level of DPA and/or DHA in the total fatty acid content of the extracted lipid is between about 1% and 35%, or between about 7% and 35% or between about 20.1% and 35%.

7. The process of claim 5, wherein the plant part has one or more or all of the following features

i) the efficiency of conversion of oleic acid to DPA and/or DHA in the plant part is at least about 10%, at least about 15%, at least about 20%, at least about 25%, about 20%, about 25%, about 30%, between about 10% and about 50%, between about 10% and about 30%, between about 10% and about 25%, or between about 20% and about 30%,

ii) the efficiency of conversion of LA to DPA and/or DHA in the plant part is at least about 15%, at least about 20%, at least about 22%, at least about 25%, at least about 30%, about 25%, about 30%, about 35%, between about 15% and about 50%, between about 20% and about 40%, or between about 20% and about 30%, and

iii) the efficiency of conversion of ALA to DPA and/or DHA in the plant part is at least about 17%, at least about 22%, at least about 24%, at least about 30%, about 30%, about 35%, about 40%, between about 17% and about 55%, between about 22% and about 35%, or between about 24% and about 35%.

8. The process of claim 5, wherein the total oil content of the plant part is at least about 40%, at least about 50%, at least about 60%, at least about 70%, between about 50% and about 80%, or between about 80% and about 100% of the total oil content of a corresponding wild-type plant part.

9. The process of claim 5 which further comprises treating the lipid to increase the level of DPA and/or DHA as a percentage of the total fatty acid content, wherein the treatment comprises one or more of fractionation, distillation or transesterification.

10. The process of claim 9 which comprises the production of methyl- or ethyl-esters of DPA and/or DHA.

11. A method of treating a lipid to increase the level of DPA and/or DHA as a percentage of the total fatty acid content, the method comprising one or more of fractionating, distillating or transesteriflying plant lipid comprising fatty acids in an esterified form, the fatty acids comprising palmitic acid and C22 polyunsaturated fatty acid which comprises docosapentaenoic acid (DPA) and/or docosahexaenoic acid (DHA), and optionally myristic acid, wherein at least 35% of the DPA and/or DHA esterified in the form of triacylglycerol (TAG) is esterified at the sn-2 position of the TAG, wherein the level of palmitic acid in the total fatty acid content of the extracted lipid is between about 2% and 16%, and wherein the level of myristic acid (C14:

0) in the total fatty acid content of the extracted lipid, if present, is less than 1%.

12. The process of claim 11 which comprises the production of methyl- or ethyl-esters of DPA and/or DHA.