CN108265090B

CN108265090B - Preparation method of antarctic krill oil substitute

Info

Publication number: CN108265090B
Application number: CN201611254242.9A
Authority: CN
Inventors: 王翔宇; 李世磊; 孔录; 刘孟涛; 王满意; 张建华; 董巍; 郝克非; 董华; 尚刚
Original assignee: Cofco Donghai Grain And Oil Industry Zhangjiagang Co ltd; Cofco Excel Joy Tianjin Co ltd; Cofco Corp; Cofco Nutrition and Health Research Institute Co Ltd
Current assignee: Cofco Donghai Grain And Oil Industry Zhangjiagang Co ltd; Cofco Excel Joy Tianjin Co ltd; Cofco Corp; Cofco Nutrition and Health Research Institute Co Ltd
Priority date: 2016-12-30
Filing date: 2016-12-30
Publication date: 2021-06-15
Anticipated expiration: 2036-12-30
Also published as: CN108265090A

Abstract

The invention discloses a preparation method of an antarctic krill oil substitute. The present invention provides a method for producing a fat composition containing phospholipid-type DHA and phospholipid-type EPA, and a fat composition produced by the method. Specifically, the invention precisely controls the proportion of DHA and EPA in the reaction raw materials to prepare the grease composition containing phospholipid DHA and phospholipid EPA which are almost the same as natural antarctic krill oil.

Description

Preparation method of antarctic krill oil substitute

Technical Field

The invention belongs to the field of preparing lipids by utilizing biocatalysis, and particularly relates to a method for preparing an antarctic krill oil substitute by using a biological enzyme preparation as a catalyst and using soybean lecithin, EPA or esterified derivatives thereof and DHA or esterified derivatives thereof as raw materials.

Background

Polyunsaturated fatty acid (PUFA) refers to a class of fatty acids with carbon number not less than 18 and containing more than two double bonds; among them, PUFA in which the first double bond is present at the 3 rd position on the carbon chain methyl group is referred to as n-3polyunsaturated fatty acid. Typical examples of n-3polyunsaturated fatty acids include docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). A large number of pharmacological and animal experiments prove that DHA is necessary for normal development of retina and normal function of nervous system. In the brain and retina, DHA represents about 20% and about 35% of the total fatty acids present therein, respectively, and is present mainly in the form of phospholipids. The data indicate that DHA accumulates in the nerve cells of the fetal central nervous system from 26-40 weeks gestation; the DHA content in the brain and cerebellum of the fetus in the last 3 months of pregnancy increased three to five times compared to 26 weeks of pregnancy. The increase of the intake of DHA in the infant period obviously promotes the growth of nerve cells and the formation of dendrite in the brain, thereby being helpful for improving the learning and memory ability. Moreover, DHA is also beneficial to preventing senile dementia. Research shows that compared with healthy elderly, DHA content of brain hippocampus cells of senile dementia patients is reduced by nearly 10%; administration of DHA may help to alleviate the symptoms. In addition, the fatty acid chain length and the degree of unsaturation are closely related to the liquidity, fluidity, refractivity, and permeability of the photoreceptor membrane in the retina. When the supply of DHA is insufficient, the photosensitivity and vision of eyes are obviously reduced. On the other hand, EPA plays an important role in immune and inflammatory responses as a polyunsaturated fatty acid chemical messenger. Specifically, EPA can exert antithrombotic effects by regulating the production of Phosphatidylglycerol (PG) at platelets and vascular walls; PGE2 synthesized via the arachidonic acid pathway is also reduced by competitive inhibition, thereby reducing the acute inflammatory response. Animal experiments show that EPA can significantly reduce blood triglyceride levels. In addition, co-administration of DHA and EPA has certain effects on the treatment of childhood hyperkinetic syndrome, the control of aggressive and violent behaviors in school-age children, and the improvement of depression, anxiety and panic in adults. All major nutrition agencies in the world have suggested DHA and EPA intake. For example, the EPA and DHA intake recommended by the food and agriculture organization and the world health organization of the united nations is 250 mg/day for adults; the intake of EPA and DHA for pregnant and lactating women is 300 mg/day. The European food safety agency also recommends that adults take 250mg EPA and DHA per day.

The above-mentioned physiological effects of DHA and EPA are well recognized by researchers and consumers worldwide. However, there are still many problems with preparing commercial DHA or EPA dietary supplements. In particular, the commercialized DHA and EPA exist mainly in the form of ethyl ester or triglyceride, and are still not desirable in terms of absorption efficiency and oxidation stability. Mouse experiments show that the rate of DHA or EPA absorption by organs such as brain, liver and kidney in the form of ethyl ester or triglyceride is more than two times lower than that of DHA or EPA in the form of phospholipid. In addition, triglycerides are used as energy sources, usually after digestion and absorption in vivo either directly into energy metabolic pathways for ATP production or transported to adipocytes as energy stores. Thus, DHA or EPA in triglyceride form is only used as a fuel and does not exert its functional advantages. In contrast, polyunsaturated fatty acids in phospholipid form occur naturally in membrane structures in the body, and DHA or EPA in phospholipid form is more likely to exert its biological activity.

Antarctic krill oil is a new food raw material approved by the Ministry of health, is the only product which is commercialized at present and naturally contains phosphatide type n-3polyunsaturated fatty acid, and is the most effective dietary supplement of DHA and EPA. However, Antarctic krill exists only in Antarctic waters and is not readily available in China. On the other hand, antarctic krill oil extracted as krill has potential sensitization. The invention aims to utilize a biological catalysis technology to connect DHA and EPA into soybean phospholipid so as to produce a substitute product with phospholipid type DHA and EPA contents completely the same as that of antarctic krill oil. The effect of the substitute product as a DHA and EPA dietary supplement is consistent with that of Antarctic krill oil; and because the allergen does not contain other allergens, the application range of people is wider.

As shown in references 1 to 7, various methods have been developed in the art for incorporating n-3polyunsaturated fatty acids into phospholipids via transesterification.

[ reference 1] Sunmamemin et al, a process for producing an n-3polyunsaturated fatty acid type phospholipid by an enzymatic method, oil and fat of China.

[ reference 2] Xiang Li et al, Production of Structured Phosphophile with High Content of DHA/EPA by Immobilized Phospholipase A1-Catalyzed Transisterication, int.J.mol.Sci.

[ reference 3] In-Hwan Kim et al, Synthesis of Structured phospholipid binding n-3PUFA resins via acids catalysis by Mediated Immobilized phospholipid A1, J.Am.oil.chem.Soc.

[ reference 4] In-Hwan Kim et al, Phospholipase A1-catalyzed synthesis of phospholipds expressed In n-3 polymerized fatty acid residues, Enzyme and Microbial Technology.

[ reference 5] Hugo S.Garcia et al, entity of molecular with n-3fat acids by acidic analysis using immobilized phospholipase A1, GRASAS YACEITES.

[ reference 6] TingTing Zhao, Immobilized phospholipases A1-catalyzed modification of phospholipases with n3 unsalted fatty acid, Food Chemistry.

[ reference 7] Life Long et al, Production of structured phosphopeptides by lipase-catalyzed acidity analysis, optimization using stress surface method, Enzyme and microbiological Technology.

However, none of the methods reported in the above documents can produce a product with phospholipid type DHA and EPA content exactly the same as that of antarctic krill oil. Specifically, references 1 to 7 all directly use purified fish oil containing polyunsaturated fatty acids/fatty acid esters as a starting material. It is well known in the art that due to the difference in the spatial structure of DHA and EPA, the two differ significantly in their access efficiency in bio-enzyme catalyzed transesterification reactions. The proportion of DHA and EPA in the purified fish oil raw material is fixed, and the proportion can not be adjusted by later-stage manual blending. Therefore, the technology of directly preparing the natural fish oil cannot realize that the EPA and the DHA have compositions similar to those of the antarctic krill oil.

Further, references 1-6 all use PLA1 enzyme to catalyze the exchange reaction of phospholipids with DHA and EPA. However, the commercial PLA1 enzyme is a liquid enzyme, and no immobilization product is currently available, which requires a complicated immobilization procedure to be performed when used. Therefore, the above process is not industrially practical.

Third,

references

1 and 2 employ ethyl ester type n-3polyunsaturated fatty acid to perform transesterification with phospholipid under solvent-free conditions, however, due to limitations of miscibility of fatty acid ethyl ester and phospholipid and uniformity of reaction system, the reaction can be achieved only when the weight ratio of fatty acid ethyl ester and phospholipid is as high as 6:1 (weight ratio is approximately 14:1), and the optimal total incorporation rate is only about 25 wt%.

In addition, references 3 to 7 use free DHA and EPA as starting materials and perform transesterification with phospholipids. The arrangement improves the intersolubility of DHA and EPA with phospholipid and the uniformity of the system, thereby greatly reducing the dosage of DHA and EPA. However, it is well known that free substrates will substantially reduce the lifetime of immobilized enzymes compared to ester substrates, thereby increasing processing costs. In addition, Stine et al provided phospholipid-type or triglyceride-type DHA to each of the two groups of subjects, and when the phospholipid-type DHA was administered at a dose of 62.8% of the triglyceride-type DHA, the blood levels of the subjects reached the same DHA and EPA indices. Clinical experiments of Hansen and the like prove that the phospholipid DHA has higher absorption efficiency than the ethyl ester DHA. In this regard, since the free fatty acids are not suitable for direct administration to humans, the above processes require further purification steps to remove a large amount of free fatty acids from the product, again increasing the cost of preparation.

Currently, there is no method in the art that can accurately produce a product with a phospholipid type DHA/EPA content close to that of antarctic krill oil.

Disclosure of Invention

In one aspect, the present invention provides a method for producing a fat composition containing phospholipid-type DHA and phospholipid-type EPA, the method comprising the steps of:

(1) mixing DHA or its ester derivative and EPA or its ester derivative at room temperature to obtain oil mixture A,

wherein, the content of DHA in the DHA or the ester derivatives thereof is not less than 75 wt%, the content of EPA in the EPA or the ester derivatives thereof is not less than 75 wt%, and the adding amount of the DHA or the ester derivatives thereof and the EPA or the ester derivatives thereof is as follows: the weight ratio of DHA to EPA in the grease mixture A is (30-70) to (70-30);

(2) mixing soybean phospholipid and the oil and fat mixture A obtained in the step (1) according to the weight ratio of (10-35) to (90-65) to obtain an oil and fat mixture B, wherein the content of phospholipid in the added soybean phospholipid is not less than 75 wt%;

(3) preheating the grease mixture obtained in the step (2) to 40-70 ℃, adding distilled water and mixing, wherein the adding amount of the distilled water is 0-6 wt% of the adding amount of the enzyme preparation in the step (4);

(4) adding the immobilized lipase or phospholipase preparation into the reaction mixture obtained in the step (3), and reacting at the temperature of 40-70 ℃ to obtain a grease composition C;

wherein the content of phospholipid type DHA and phospholipid type EPA is 9.0-15.0 wt% and 18.0-25.0 wt% of the oil and fat composition C, respectively.

In another aspect, the present invention provides the fat and oil composition containing phospholipid-type DHA and phospholipid-type EPA prepared by the method according to the first aspect.

Specifically, the invention is realized by the following technical scheme:

1. a method for producing a fat composition containing phospholipid-type DHA and phospholipid-type EPA, the method comprising the steps of:

2. The method of paragraph 1, wherein in step (1), the DHA or ester derivative thereof is from any of fish oil, algal oil, microbial oil, or any combination thereof; the EPA or its ester derivative is selected from fish oil, algae oil, microbial oil, or any combination of the above oils.

3. The method according to

paragraph

1 or 2, wherein in step (1), the DHA or ester derivative thereof is selected from a methyl ester type derivative, an ethyl ester type derivative, a glyceride type derivative, or free DHA, or any combination of the aforementioned DHA or ester derivative thereof; the EPA or the ester derivative thereof is selected from methyl ester type derivatives, ethyl ester type derivatives, glyceride type derivatives or free EPA of EPA, or any combination of the EPA or the ester derivative thereof; preferably, the DHA or ester derivative thereof is a DHA ethyl ester type derivative; preferably, the EPA or ester derivative thereof is an EPA ethyl ester type derivative.

5. The method of any of paragraphs 1-4, wherein in step (1), the DHA or ester derivative thereof and EPA or ester derivative thereof are added in amounts of: the weight ratio of DHA to EPA in the grease mixture A is (35-45) to (65-55).

6. The method of any of paragraphs 1-5, wherein in step (2) the soybean phospholipids comprise phospholipids in an amount of not less than 85% by weight.

7. The method according to any one of paragraphs 1 to 6, wherein in step (2), the soybean phospholipids and the fat and oil mixture A obtained in step (1) are mixed in a weight ratio of (20-30) to (80-70).

8. The method of any of paragraphs 1-7, wherein in step (2), the mixing is performed in a solvent-free system.

9. The method of any of paragraphs 1-7, wherein in step (2), the mixing is performed in an organic solvent selected from any one or any combination of the group consisting of n-hexane, isohexane, acetone; preferably, the organic solvent is n-hexane.

10. The method of paragraph 9 wherein the n-hexane is added in an amount of: 1ml-1.5ml of n-hexane was added per 100mg of phospholipid.

11. The method of any of paragraphs 1-10, wherein in step (3), said preheating of step (hi) is at 45 ℃ -65 ℃.

12. The method of any of paragraphs 1-11, wherein in step (3) the amount of distilled water added is 1.5-3.5 wt% of the amount of enzyme preparation added in step (4).

13. The method of any of paragraphs 1-12, wherein in step (4) the immobilized lipase or phospholipase preparation is an immobilized lipase preparation.

14. The method of paragraph 13 wherein the immobilized lipase preparation is derived from one or more fungi selected from the group consisting of: rhizomucor miehei, Thermomyces lanuginosus and Aspergillus niger.

15. The method of paragraph 14 wherein the immobilized lipase preparation is a Rhizomucor miehei IM lipase preparation, a Thermomyces lanuginosus TL IM lipase preparation, or a SP435 lipase preparation from Novitin.

16. The method of any of paragraphs 1-15, wherein in step (4) the immobilized lipase or phospholipase preparation is added in an amount of 5 wt% to 35 wt%, preferably 10 wt% to 30 wt% of the fat blend B.

17. The method of any of paragraphs 1-16, wherein in step (4), the reaction temperature is from 45 ℃ to 65 ℃.

18. The method of any of paragraphs 1-17, wherein in step (4), the reaction is carried out for 1-48 hours, preferably 8-24 hours.

19. The fat and oil composition comprising phospholipid-type DHA and phospholipid-type EPA produced by the method according to any one of paragraphs 1 to 18, wherein the content of phospholipid-type DHA and phospholipid-type EPA is 9.0 to 15.0 wt% and 18.0 to 25.0 wt%, respectively, of the fat and oil composition C.

The term "room temperature" as used herein means a temperature range of ambient temperature from 16 to 26 ℃.

Advantageous effects

Due to the selectivity of fungal lipases and phospholipases for substrates, the difficulties of EPA and DHA access differ greatly. There is no report in the art of preparing a lipid composition with similar content of phospholipid type DHA and EPA as that of Antarctic krill oil. The invention firstly provides that the grease composition containing phospholipid DHA and phospholipid EPA which are almost the same as natural antarctic krill oil is prepared by accurately controlling the proportion of DHA and EPA in reaction raw materials. In the product of the present invention, the content of phospholipid-type DHA and phospholipid-type EPA is 9.0 to 15.0 wt% and 18.0 to 25.0 wt%, respectively, of the fat composition. Such a ratio helps DHA and EPA to be sufficiently absorbed and utilized in the body.

As described above, in human body, the phospholipid type n-3polyunsaturated fatty acid has a significantly higher absorption and utilization rate than the triglyceride type and the ethyl ester type n-3polyunsaturated fatty acid, and thus is more suitable for consumers who have requirements for promoting brain development and improving cardiovascular health. On the other hand, due to high absorption and utilization rate, the product of the invention can be used as a dietary supplement to reduce the total lipid intake while realizing the equivalent biological activity of the prior art, thereby being more in line with the health concept.

In addition, the invention particularly selects commercial immobilized enzyme as the catalyst, improves the repeatability and industrial feasibility of the reaction, and can realize the batch preparation of the antarctic krill oil substitute.

In a preferred embodiment, n-hexane (processing aid in GB 2760-2014) is added into the grease mixture B, so that the miscibility of the soybean lecithin and the ester containing EPA or DHA is increased, and the dosage of DHA and EPA is greatly reduced. In other preferred embodiments, the ethyl ester type derivative of DHA or EPA is used as the starting material in high yield and without the need to isolate the free fatty acids after the reaction is complete. However, the efficiency of the lipase preparation for catalyzing the esterification reaction of EPA or DHA is affected by the steric hindrance caused by the presence of multiple double bonds in EPA or DHA itself. Particularly, when ethyl ester type EPA or DHA is used, the effect of the steric hindrance is more significant, resulting in a decrease in the access rate. The inventors of the present invention found that the addition of n-hexane allows the viscosity of the reaction system to be reduced, thus improving the fluidity of the reactants, and allows the reaction of the ethyl ester-based substrate with lipase to be more easily carried out, thus reducing the reaction time. Therefore, the ethyl ester type EPA or DHA is used while the normal hexane is added into the system, so that the raw materials are saved, the reaction time is shortened, the yield is improved, and the industrial feasibility is higher.

Detailed Description

The present invention will be described in detail below.

The term "phospholipid-type DHA" or "phospholipid-type EPA" as used herein refers to a phospholipid in which DHA or EPA is grafted to the 1 or 2 position of the phosphoglyceryl moiety. As is well known in the art, phospholipids are a class of phosphate-containing lipids, generally having a hydrophilic head and a hydrophobic tail; the hydrophilic head is composed of a phosphate group and a substituent (e.g., an ammonia-containing base or an alcohol) attached to the phosphate group, and the hydrophobic tail is composed of a fatty acid chain. The hydrophilic head of phospholipid type DHA and phospholipid type EPA may be Phosphatidic Acid (PA), Phosphatidylcholine (PC), Phosphatidylethanolamine (PE), Phosphatidylglycerol (PG), Phosphatidylinositol (PI), etc.

In a first aspect, the present invention relates to a method for producing a fat composition containing phospholipid-type DHA and phospholipid-type EPA. The oil and fat composition prepared by the method has the content of phospholipid type DHA and phospholipid type EPA of 9.0-15.0 wt% and 18.0-25.0 wt% of the oil and fat composition product respectively. As demonstrated in the examples, the content of phospholipid-type DHA and phospholipid-type EPA in antarctic krill oil was about 13.9 wt% and 22.0 wt%, respectively. Therefore, the product prepared by the method can be used as an antarctic krill oil substitute for supplementing DHA and EPA.

In some embodiments, the DHA or ester derivative thereof and EPA or ester derivative thereof used in step (1) of the process of the invention may be of any origin, for example from any of fish oil, algal oil, microbial oil, or any combination of the above. The DHA or the ester derivative thereof can be methyl ester type derivative, ethyl ester type derivative, glyceride type derivative of DHA, free type DHA, or any combination of the DHA or the ester derivative thereof. The EPA or its ester derivative can be selected from methyl ester type derivative, ethyl ester type derivative, glyceride type derivative, free EPA, or any combination of the above EPA or its ester derivative. Preferably, the DHA or ester derivative thereof is a DHA ethyl ester type derivative. Preferably, the EPA or ester derivative thereof is an EPA ethyl ester type derivative. The DHA or the ester derivative thereof contains DHA not less than 75 wt%, and the EPA or the ester derivative thereof contains EPA not less than 75 wt%.

In the step (1) of the present invention, the ratio of DHA and EPA in the fat mixture a needs to be precisely controlled, so that the content of phospholipid DHA and phospholipid EPA in the obtained product is close to that of the antarctic krill oil. Preferably, the DHA or ester derivative thereof and EPA or ester derivative thereof are added in an amount of: the weight ratio of DHA to EPA in the grease mixture A is (35-45) to (65-55).

Soybean phospholipids are commonly used as industrial raw materials in the field, and mainly contain phosphatidic acid, phosphatidylcholine (also called lecithin), phosphatidylethanolamine and phosphatidylinositol, wherein the content of phosphatidylcholine is the highest. The content of phospholipids in the soybean phospholipids used in the step (2) of the method of the present invention is preferably not less than 75% by weight, more preferably not less than 85% by weight. The effect of the present invention can be achieved when the mixing ratio of the soybean phospholipids and the fat and oil mixture A obtained in the step (1) is (10-35): (90-65), however, the mixing ratio is more preferable to (20-30): (80-70).

The mixing of step (1) and step (2) of the present invention is carried out at room temperature (generally 16 to 26 ℃). The mixing is carried out in the absence of a solvent system or in the presence of an organic solvent. In order to increase the miscibility of the phospholipids with the DHA/EPA containing esters, it is preferred to perform the mixing of step (2) in the presence of an organic solvent. The organic solvent is selected from any one or any combination of n-hexane, isohexane and acetone; preferably, the organic solvent is n-hexane. The adding amount of the normal hexane is determined according to the content of the phospholipid in the reaction mixture. Preferably, the addition amount of the n-hexane is as follows: 1ml-1.5ml of n-hexane was added per 100mg of phospholipid.

The preheating temperature of the fat mixture B in step (3) of the process of the invention is from 40 ℃ to 70 ℃, preferably from 45 ℃ to 65 ℃. The amount of distilled water to be added needs to be precisely controlled, and is preferably 0 to 6 wt%, more preferably 1.5 to 3.5 wt% of the amount of the enzyme preparation to be added in step (4).

In step (4) of the process of the invention, the immobilized lipase or phospholipase preparation is an immobilized lipase preparation, preferably an immobilized lipase preparation. Preferably, the immobilized lipase preparation is derived from one or more fungi selected from the group consisting of: rhizomucor miehei (Rhizomucor miehei), Thermomyces lanuginosus (Thermomyces lanuginosus), and Aspergillus niger (Aspergillus niger). Preferably, the immobilized lipase preparation is a Rhizomucor mie IM lipase preparation, a Thermomyces lanuginosus TL IM lipase preparation or a SP435 lipase preparation from Novistin. In some embodiments, the immobilized lipase or phospholipase preparation is added in an amount of 5 wt% to 35 wt%, preferably 10 wt% to 30 wt% of the fat blend B. The reaction temperature is controlled at 40 ℃ to 70 ℃, preferably 45 ℃ to 65 ℃. The reaction is carried out for 1 to 48 hours, preferably 8 to 24 hours.

In a second aspect, the present invention relates to the fat and oil composition containing phospholipid-type DHA and phospholipid-type EPA prepared by the method according to the first aspect, wherein the content of phospholipid-type DHA and phospholipid-type EPA is 9.0 to 15.0 wt% and 18.0 to 25.0 wt%, respectively, of the fat and oil composition C.

Drawings

Fig. 1 is a gas chromatogram obtained by analyzing the composition of all fatty acids in a fat standard sample by chromatography.

Fig. 2 is a gas chromatogram of the oil or fat composition 1 obtained in example 1 of the present invention.

Fig. 3 is a gas chromatogram of the oil or fat composition 1 obtained in comparative example 1 of the present invention.

Examples

The present invention will be described in further detail with reference to examples. These examples are merely illustrative and should not be construed as limiting the scope of the invention. All technical solutions and modifications thereof implemented based on the above contents of the present invention fall within the scope of the present invention.

The phospholipids used in examples and comparative examples were purchased from Sigma-Aldrich (product No. 1001900076), and the content of the phospholipids in the soybean phospholipids was 86 wt%. The DHA ester derivative and the EPA ester derivative are respectively purchased from BASF (China) Limited company (concentrated DHA (ethyl ester type), 50382353) and Sn-free Xunda marine biological products Limited company (concentrated EPA (ethyl ester type, EPE150818) wherein the DHA content in the DHA ester derivative is 79.7 wt% and the EPA content in the EPA ester derivative is 76.2 wt%, the Antarctic krill oil is purchased from Wei Aker company (0995250100), immobilized RM IM lipase, PLA1 lipase (liquid state), TL IM lipase (immobilized) and SP435 lipase (immobilized) are all purchased from Norwesson (China) biotechnology Limited company (product numbers are CHE-20, LY05060, LA331342 and LC200266 respectively), and n-hexane is analytically pure and purchased from national medicine group.

In the examples and comparative examples, the distribution and content of fatty acids in each sample were analyzed by gas chromatography according to the national standard GB 28404-2012, using a gas chromatograph model Agilent 1890B.

Specifically, the compositions of all the fatty acids in the oil and fat standard samples (fatty acid methyl ester mix, 37 kinds of C4-C24, product No. 18919-1AMP from sigma Co.) were analyzed by chromatography (FIG. 1). And comparing with a standard spectrum to obtain the distribution and content of each fatty acid in the test sample. Tables 1 to 2 below show the contents of respective fatty acids in the raw material of the present invention, DHA ester derivatives, EPA ester derivatives and soybean phospholipids, measured by chromatography. Table 3 below lists the content of each fatty acid in the commercially available antarctic krill oil, as determined by chromatography. The content of each fatty acid in the fat and oil composition C obtained in each of the examples and comparative examples was measured by the method described above. It is noted that the measured values of the percentage content of DHA or EPA shown in table 4 are the DHA or EPA access rates.

TABLE 1 content of respective fatty acids in soybean lecithin (% by weight)

TABLE 2 DHA ester derivatives and EPA ester derivatives content of respective fatty acids

	DHA ester derivative (wt%)	EPA ester derivatives (wt%)
			C16:0	1.1	1.6
C16:1	0.7	0.3
			C18:0	0.3	0.6
C18:1	1.0	0.4
			C18:2	0.3	0.2
C20:1	0.1	0.1
			C21:0	0.2	0.1
C20:4	1.8	2.0
			C23:0	0.5	0.3
C20:5	8.8	76.2
			C24:0	0.3	0.1
C24:1	1.0	1.3
			C22:6	79.7	10.2
Is unknown	4.2	6.2

TABLE 3 content of fatty acids in Antarctic krill oil (wt%)

Examples 1 to 8 preparation of fat and oil compositions containing phospholipid-type DHA and phospholipid-type EPA in amounts similar to Antarctic krill oil

[ oil and fat composition 1]

(1) Weighing 245mg of DHA ester derivative and 455mg of EPA ester derivative, adding the DHA ester derivative and the EPA ester derivative into a reactor at room temperature, and fully mixing to obtain a grease mixture A;

(2) weighing 300mg of soybean phospholipid, adding the soybean phospholipid and 3ml of n-hexane into a reactor, and fully mixing the soybean phospholipid and the n-hexane with the grease mixture A obtained in the step (1) to obtain a grease mixture B;

(3) preheating the grease mixture B obtained in the step (2) to 55 ℃, adding precisely measured 4.5 mu l of distilled water into the grease mixture B, and fully and uniformly mixing;

(4) adding 300mg of RM IM lipase preparation to the reaction mixture obtained in the step (3), and reacting at 55 ℃ for 16 hours to obtain the grease composition 1.

The gas chromatogram of the oil or fat composition 1 is shown in fig. 2.

[ oil and fat composition 2]

(1) Weighing 315mg of DHA ester derivative and 385mg of EPA ester derivative, adding the DHA ester derivative and the EPA ester derivative into a reactor at room temperature, and fully mixing to obtain a grease mixture A;

(4) adding 300mg of RM IM lipase preparation to the reaction mixture obtained in the step (3), and reacting at 55 ℃ for 16 hours to obtain the oil composition 2.

[ oil and fat composition 3]

(1) Weighing 280mg of DHA ester derivative and 520mg of EPA ester derivative, adding the DHA ester derivative and the EPA ester derivative into a reactor at room temperature, and fully mixing to obtain a grease mixture A;

(2) weighing 200mg of soybean phospholipid, adding the soybean phospholipid and 3ml of n-hexane into a reactor, and fully mixing the soybean phospholipid and the n-hexane with the grease mixture A obtained in the step (1) to obtain a grease mixture B;

(4) adding 300mg of RM IM lipase preparation into the reaction mixture obtained in the step (3), and reacting at the temperature of 55 ℃ for 16 hours to obtain the grease composition 3.

[ oil and fat composition 4]

(4) 300mg of TL IM lipase preparation was added to the reaction mixture obtained in the step (3) and reacted at a temperature of 55 ℃ for 16 hours to obtain a fat composition 4.

[ oil and fat composition 5]

(4) 300mg of the SP435 lipase preparation was added to the reaction mixture obtained in step (3) and reacted at a temperature of 55 ℃ for 16 hours to obtain an oil or fat composition 5.

[ oil and fat composition 6]

(3) preheating the grease mixture B obtained in the step (2) to 55 ℃, adding precisely measured 5.25 mu l of distilled water into the grease mixture B, and fully and uniformly mixing;

(4) adding 300mg of RM IM lipase preparation to the reaction mixture obtained in the step (3), and reacting at 55 ℃ for 16 hours to obtain the grease composition 6.

[ oil and fat composition 7]

(1) Weighing 245mg of DHA ester derivative and 525mg of EPA ester derivative, adding the DHA ester derivative and the EPA ester derivative into a reactor at room temperature, and fully mixing to obtain a grease mixture A;

(4) adding 300mg of RM IM lipase preparation to the reaction mixture obtained in the step (3), and reacting at 55 ℃ for 24 hours to obtain the grease composition 7.

[ oil and fat composition 8]

(4) adding 300mg of RM IM lipase preparation to the reaction mixture obtained in the step (3), and reacting at 55 ℃ for 8 hours to obtain the oil composition 8.

Comparative examples 1 to 3: preparation of an oil and fat composition containing phospholipid-type DHA and phospholipid-type EPA contents [ oil and fat composition c1]

(1) Weighing 520mg of DHA ester derivative and 180mg of EPA ester derivative, adding the DHA ester derivative and the EPA ester derivative into a reactor at room temperature, and fully mixing to obtain a grease mixture A;

(4) adding 300mg of RM IM lipase preparation into the reaction mixture obtained in the step (3), and reacting at the temperature of 55 ℃ for 16 hours to obtain the grease composition c 1.

The gas chromatogram of the oil or fat composition c1 is shown in FIG. 3.

[ oil and fat composition c2]

(1) Weighing 125mg of DHA ester derivative and 575mg of EPA ester derivative, adding the DHA ester derivative and the EPA ester derivative into a reactor at room temperature, and fully mixing to obtain a grease mixture A;

(4) adding 300mg of RM IM lipase preparation into the reaction mixture obtained in the step (3), and reacting at the temperature of 55 ℃ for 16 hours to obtain the grease composition c 2.

[ oil and fat composition c3]

(2) weighing 400mg of soybean phospholipid, adding the soybean phospholipid and 3ml of n-hexane into a reactor, and fully mixing the soybean phospholipid and the n-hexane with the grease mixture A obtained in the step (1) to obtain a grease mixture B;

(4) adding 300mg of RM IM lipase preparation into the reaction mixture obtained in the step (3), and reacting at the temperature of 55 ℃ for 16 hours to obtain the grease composition c 3.

[ oil and fat composition c4]

(2) weighing 300mg of soybean phospholipid, adding the soybean phospholipid into a reactor, and fully mixing the soybean phospholipid with the grease mixture A obtained in the step (1) to obtain a grease mixture B;

(4) adding 300mg of RM IM lipase preparation into the reaction mixture obtained in the step (3), and reacting at the temperature of 55 ℃ for 16 hours to obtain the grease composition c 4.

[ oil and fat composition c5]

(3) preheating the grease mixture B obtained in the step (2) to 30 ℃, adding precisely measured 4.5 mu l of distilled water into the grease mixture B, and fully and uniformly mixing;

(4) adding 300mg of RM IM lipase preparation into the reaction mixture obtained in the step (3), and reacting at the temperature of 30 ℃ for 16 hours to obtain the grease composition c 5.

[ oil and fat composition c6]

(3) preheating the grease mixture B obtained in the step (2) to 80 ℃, adding precisely measured 4.5 mu l of distilled water into the grease mixture B, and fully and uniformly mixing;

(4) adding 300mg of RM IM lipase preparation into the reaction mixture obtained in the step (3), and reacting at the temperature of 80 ℃ for 16 hours to obtain the grease composition c 6.

TABLE 4

Claims

(4) adding an immobilized lipase preparation into the reaction mixture obtained in the step (3), and reacting at the temperature of 40-70 ℃ to obtain a grease composition C, wherein the lipase preparation is a Rhizomucor miehei RMIM lipase preparation, a Thermomyces lanuginosus TLIM lipase preparation or an SP435 lipase preparation from Novistin;

2. The method according to claim 1, wherein in step (1), the DHA or the ester derivative thereof is derived from any one of fish oil, algae oil, microbial oil, or any combination of the above oils; the EPA or its ester derivative is selected from fish oil, algae oil, microbial oil, or any combination of the above oils.

3. The method according to claim 1 or 2, wherein in step (1), the DHA or ester derivative thereof is selected from a methyl ester type derivative, an ethyl ester type derivative, a glyceride type derivative or free DHA of DHA, or any combination of the aforementioned DHA or ester derivative thereof; the EPA or its ester derivative is selected from methyl ester type derivative, ethyl ester type derivative, glyceride type derivative or free EPA of EPA, or any combination of the above EPA or its ester derivative.

4. The method according to claim 3, wherein the DHA or ester derivative thereof is an ethyl ester type DHA derivative and the EPA or ester derivative thereof is an ethyl ester type EPA derivative.

5. The method according to claim 1 or 2, wherein the DHA or ester derivative thereof and EPA or ester derivative thereof are added in step (1) in amounts of: the weight ratio of DHA to EPA in the grease mixture A is (35-45) to (65-55).

6. The method according to claim 1 or 2, wherein the soybean phospholipid is contained in an amount of not less than 85 wt% in the step (2).

7. The method according to claim 1 or 2, wherein in the step (2), the soybean phospholipids and the fat and oil mixture A obtained in the step (1) are mixed in a weight ratio of (20-30) to (80-70).

8. The process according to claim 1 or 2, wherein in step (2), the mixing is carried out in a solvent-free system.

9. The method according to claim 1 or 2, wherein in the step (2), the mixing is performed in an organic solvent selected from any one or any combination of the group consisting of n-hexane, isohexane, acetone.

10. The method of claim 9, wherein the organic solvent is n-hexane.

11. The method of claim 10, wherein the n-hexane is added in an amount of: 1ml-1.5ml of n-hexane was added per 100mg of phospholipid.

12. The method of claim 1 or 2, wherein in step (3), the preheating is performed at 45 ℃ to 65 ℃.

13. The method according to claim 1 or 2, wherein the distilled water is added in an amount of 1.5-3.5 wt% of the amount of the enzyme preparation added in step (4) in step (3).

14. The method according to claim 1 or 2, wherein in the step (4), the lipase preparation is added in an amount of 5 wt% to 35 wt% of the fat and oil mixture B.

15. The method according to claim 14, wherein in the step (4), the lipase preparation is added in an amount of 10 wt% to 30 wt% of the fat and oil mixture B.

16. The process of claim 1 or 2, wherein in step (4), the reaction temperature is 45 ℃ to 65 ℃.

17. The method according to claim 1 or 2, wherein, in the step (4), the reaction is carried out for 1 to 48 hours.

18. The method of claim 17, wherein the reaction is carried out for 8 to 24 hours in step (4).