KR20010010268A - Arachidonic Acid Reinforced Feed Composition - Google Patents

Abstract

PURPOSE: A feed composition for a layer containing Mortierella alpina producing arachidonic acid is provided, to increase an arachidonic acid content in the yolk by cultivating Mortierella alpina and adding the dried biomass to a feed for a layer. CONSTITUTION: Feed composition enriched with arachidonic acid contains Mortierella alpina whose amount is 0.5-15 wt.% of a feed. The content of total lipid in the Mortierella alpina is 20-50 wt.% and the content of arachidonic acid in total lipid is 20-50 wt.%. The content of arachidonic acid in eggs is accumulated in a range of 2-10 wt.% in yolk lipid. The content of arachidonic acid in egg lecithin is extremely small, and Mortierella alpina is cultivated for producing and accumulating archidonic acid in the egg lecithin by adding to a feed. The feed composition enriched with arachidonic acid is used for manufacturing unrefined powdered milk in a lecithin form.

Description

Arachidonic Acid Reinforced Feed Composition

The present invention relates to a feed composition for laying hens to which a dry cell of Mortierella alpina is added and a method for producing arachidonic acid-enriched eggs produced by feeding the feed composition.

In the past, most of the eggs sold on the market were simply used as food for nourishment, but recently, the desired nutrients, elements or substances are mixed with the feed for laying hens to produce eggs containing a large amount of the ingredients. Efforts have been made to allow the human body to consume the ingredients through eggs. For example, a method of producing eggs with low cholesterol by feeding the hens a diet mainly containing seaweed or blue fish is used. Korean Patent Laid-Open Publication No. 92-3981 discloses eggs containing a large amount of iodine effective for the treatment and prevention of inflammatory diseases. In addition, Korean Patent Publication No. 95-13949 discloses omega-3 unsaturated fatty acids from laying hens by mixing the resulting oil containing omega-3 unsaturated fatty acids in the feed and feeding the mixed feed to the laying hens. A method for preparing a large amount of eggs is described. In addition, eggs containing a large amount of vitamins, eggs containing ginseng components, eggs containing components of seaweed, etc. are commercially available in the market.

Arachidonic acid is a polyunsaturated fatty acid (5, 8, 11, 14-position) having 4 cis double bonds in the ω-6 series having 20 C6, 20: 4 carbon atoms (5, 8, 11, 14 Eicosadechienophosphate) is not produced in mammals itself, but is an essential fatty acid, linolenic acid or alpha-linolenic acid (α- It is produced through the synthesis pathway of gamma linolenic acid and dihomo-gamma-linolenic acid from linolenic acid and is one of the major components of cell membrane. In addition, arachidonic acid is a precursor of prostaglandin, leukotriens, and thromboxane produced in the arachidonic acid cascade, and these eicosanoids are developed, platelet aggregation inhibitory, and plasma It is a bioactive substance with important functions involved in lowering cholesterol, regulating blood and immune system, and protecting skin.

The elderly, the sick, infants, and infants have low enzyme activity such as delta6-desaturase and delta5-desaturase known as enzymes involved in arachidonic acid biosynthesis. Because of the low biosynthesis of arachidonic acid, dietary supply of arachidonic acid is necessary. (Shimizu, S., Kawashima, H. (1993) Japan 42 (4): 254-264.)

Arachidonic acid is an essential ingredient for early growth and brain development of infants. It is the most abundant fatty acid in breast milk, and it is essential to add arachidonic acid to the formula to make infant formula close to breast milk. In order to breast-feed infant formula such as infant formula, mass supply of arachidonic acid is required, and as the transition to an aging society increases interest in maintaining health, the market for health food containing arachidonic acid is rapidly increasing. .

Arachidonic acid is currently extracted from animal tissues such as liver, sardine, and egg yolk in pigs, but its content is so low that the extraction cost is excessive, making it difficult to use as an economical source of arachidonic acid.

However, the method of producing arachidonic acid using microorganisms uses cultured microorganisms, so it is easy and economical to extract arachidonic acid from cultured cells because the supply and demand of raw materials is stable and the content of arachidonic acid in cells is constant and accumulated at high concentration. The method is being sought and is much more advantageous in terms of stable supply of raw materials.

At present, the formula is commercially available in accordance with the trend of breast milking of the formula is adding a variety of useful substances, including arachidonic acid. However, until now, the use of microorganism-derived arachidonic acid in the formula has been controversial and it has not been used. The formula is using egg lecithin as a raw material of arachidonic acid.

However, since the content of arachidonic acid in egg yolk lecithin is very small, it was necessary to develop a method for increasing the content of arachidonic acid in lecithin in order to add sufficient arachidonic acid to the formula.

The present inventors intended to increase the content of arachidonic acid in egg yolk in order to increase the content of arachidonic acid in egg yolk lecithin, and for this purpose, arachidone is produced by culturing a motiella which is a microorganism that produces arachidonic acid and accumulates in cells. Acid was to accumulate in egg yolk.

The method of the present invention does not go through a complicated and expensive process of extracting and purifying arachidonic acid from microorganisms, and culturing motiella, which is a microorganism that produces arachidonic acid and accumulates in cells, and adds arachidonic acid to egg laying feed. It is economical and simpler to use than the existing process by adopting the accumulation method inside. In addition, since it can finally be used in formula in the form of lecithin, there is an advantage that can be directly applied to the manufacturing process without objection to the use of microorganism-derived arachidonic acid.

[Preparation of Motiella Dry Cells Containing Large Arachidonic Acid]

First, the present inventors used the Motiella genus strain as a microorganism that produces arachidonic acid and accumulates in cells. This strain was Motier, a microorganism deposited by the Applicant to the Gene Bank of the Biotechnology Research Institute, attached to the Korea Institute of Science and Technology at the time of patent application (patent application number 97-57929) on November 4, 1997 (microbial accession number KCTC 8832P). Lella genus MM-25 strain was used.

The following method was used for culturing Motiella strains, which are arachidonic acid producing microorganisms. The culture was carried out using agitation conditions of 25 ℃, pH 5 ~ 5.5, 0.5vvm using a 300L medium added with glucose 50g / L, CSL 35g / L, agitation rate 250rpm. The total incubation period was 6 days and the cell mass of 17.2 g / L was obtained after the incubation. The total lipid content reached 45% of the cell mass at the end of the culture, and the content of arachidonic acid reached 40% of the total lipid content to obtain 2.5 g of arachidonic acid (3.096 g / L of dry cell basis) per liter of culture. After culturing, the culture medium was recovered by using a compression filter, and then dried under hot air at 60 ° C. for 5 days, and then pulverized to obtain a fine powdery microbial dry cell.

EXAMPLE

Preparation of Laying Hen Feed Compositions with Motiella Alpina Dry Cells

The laying hen composition for accumulating in the egg yolk of arachidonic acid prepared the composition with the following component and compounding ratio.

The dry cells added to the feed composition of the present invention is Mothierella alpina, it is most preferably used within the range of 0.5 to 15% by weight of the feed. This is because when the dry cells are added to the feed in the above range, arachidonic acid in egg yolks in laying hens is most effective for economic efficiency while maintaining an effective balance with other nutrients. If it is added below 0.5% by weight, the accumulation of arachidonic acid in egg yolk is insignificant, and if it is added more than 15%, the ratio of arachidonic acid in yolk may increase, but it is not only difficult to balance with other nutrients in other feeds. Costly and not economical.

On the other hand, the dry cells used in the present invention, it is preferable to use dry cells having a lipid content of 20 to 50% by weight in the dry cells and 20 to 50% by weight of the arachidonic acid content in the lipid.

The dry cells added to the feed composition of the present embodiment was about 45% by weight of the lipid content in the dry cells, dry cells containing 40% by weight of arachidonic acid in the lipid was used in the feed composition.

In Examples 1 to 5 of the present invention, a feed containing 0.89% to 8.89% by weight of dry cells of the feed was administered to the laying hen, and when the fatty acid in egg yolk was analyzed after the feed composition was administered, it was in the range of 3 to 10% by weight. Arachidonic acid was confirmed to accumulate (see Table 5).

▶ Feed composition manufacture

(unit %)

Comparing only the lipid content of the dry cells excluding the composition of the basic blended feed is shown in Table 1 below.

Table 1.Table comparing only lipid content of dry cells except basic blended feed composition

(Unit: weight%)

division Soybean oil Dry Cells (Based on Total Lipid Content) Comparative Example 1 (ARA 0.4%) Example 2 (ARA 0.8%) Example 3 (ARA 1.6%) Example 4 (ARA 2.4%) Example 5 (ARA 4.0%) 4.0 3.6 3.2 2.4 1.6 0.0 0 0.4 0.8 1.6 2.4 4.0

* Note: ARA is Arachidonic Acid

(Test Example) Arachidonic Acid Accumulation Experiment in Egg Yolk by Feed Composition Feeding

The laying hens used in the experiment were 24 29-week-old brown laying hens (breed: Isa Brown) purchased from a general poultry farm. Breeding conditions were the temperature 20 1 ℃, humidity 50 5%, and the illumination was illuminated using a flasher for 16 hours of 24 hours.

Experimental laying hens were administered the Comparative Example feed during the adaptation period of 10 days after the introduction, and starting from the 11th day the feed prepared for each experimental group (Comparative Example 1, Examples 1, 2, 3, 4, 5) Administered. Samples for egg yolk analysis were taken every 4 to 5 days by administering experimental feed for 4 weeks after feed replacement. During the entire experiment period, the number of eggs, egg production rate and egg weight, which were related to egg production, were recorded to observe the change of egg production according to feed change. Egg yolk was stored in -70 ℃ freezer after separation from egg white and analyzed after the experiment.

No change in egg weight was found to increase or decrease by feed in all groups (Table 2), and egg production was not changed (Table 3). Also, the ratio of egg yolk weight to egg weight was not changed in all groups. Appeared without significant difference (Table 4) it was confirmed that there is no effect on the spawning of the laying hen following the administration of dry cells.

Fatty acid changes in egg yolk increased as arachidonic acid, 16: 1, 16: 0, and 18: 3 and 18: 2, as shown in Table 5 below. In particular, arachidonic acid was significantly increased depending on the amount of cells administered. In addition, arachidonic acid was found to accumulate in the sixth day of administration.

Table 2. Changes in weight during administration

(Unit: g)

Before administration 6 days 10 days 15th 20 days 24 days 29 days 34 days 40 days Comparative example 61.3 60.2 59.9 60.0 63.1 61.7 62.0 59.7 62.5 Example 1 61.9 60.7 62.2 60.0 59.4 58.9 60.0 59.6 59.1 Example 2 59.3 58.2 59.8 59.3 59.1 61.0 62.7 62.7 61.2 Example 3 56.9 58.1 58.6 58.6 58.5 58.5 57.8 57.7 58.3 Example 4 58.3 56.8 58.3 57.4 59.1 60.8 60.7 60.9 61.0 Example 5 61.3 62.1 60.7 60.4 61.2 62.3 61.5 62.4 61.9

* Note: The figures above are cumulative averages over time

Table 3. Changes in Egg Production Rate with Feed Composition

(Unit: g)

Before administration 6 days 10 days 15th 20 days 24 days 29 days 34 days 40 days Comparative example 56.3 91.7 75.0 95.0 70.0 62.5 90.0 95.0 91.7 Example 1 81.3 87.5 88.8 85.0 95.0 81.3 90.0 95.0 91.7 Example 2 85.7 91.7 87.5 85.0 85.0 88.7 90.0 91.5 93.8 Example 3 93.8 104.2 93.8 95.0 95.0 93.8 95.0 100.0 91.7 Example 4 93.8 100.0 81.3 105.0 95.0 100.0 95.0 95.0 95.8 Example 5 87.5 95.8 81.3 85.0 95.0 93.8 90.0 90.0 100.0

Table 4. Ratio of yolk weight to yolk

Before administration 5 days after dosing 9 days after dosing 14 days after dosing 19 days after dosing 23 days after dosing 28 days after dosing Comparative example 23.4 22.5 23.8 24.5 24.5 23.3 23.8 Example 1 24.1 22.7 22.7 23.6 23.5 23.5 23.3 Example 2 22.5 22.4 22.1 22.3 23.5 22.8 23.3 Example 3 25.2 24.1 24.3 24.8 25.4 24.6 23.7 Example 4 24.8 25.1 24.1 23.9 24.5 24.8 24.8 Example 5 24.0 23.7 23.7 24.0 23.6 23.9 23.2

Table 5. Results of Analysis of Fatty Acids in Egg Yolk after Feed Composition Administration

(weight %)

6 days after dosing 14:00 16:00 16: 1 18: 0 18: 1 18: 1 isomer 18: 2 18: 3 ARA EPA DHA Comparative example 0.25 20.99 1.92 6.13 41.97 1.63 21.87 0.81 2.02 0.00 0.47 Example 1 0.24 22.09 1.46 10.88 33.82 1.57 21.89 0.86 3.07 0.00 0.99 Example 2 0.25 22.24 1.19 10.52 34.47 1.26 22.68 0.75 3.18 0.00 0.81 Example 3 0.29 23.69 1.80 10.11 34.11 1.33 20.43 0.69 3.32 0.00 0.73 Example 4 0.30 23.27 1.37 9.92 35.61 1.28 19.90 0.72 3.70 0.00 0.99 Example 5 0.35 24.29 1.48 9.93 34.44 1.31 18.45 0.53 4.59 0.00 0.74

15 days after dosing 14:00 16:00 16: 1 18: 0 18: 1 18: 1 isomer 18: 2 18: 3 ARA EPA DHA Comparative example 0.25 20.46 1.27 9.32 36.38 1.54 23.58 0.88 2.21 0.00 0.71 Example 1 0.25 22.11 1.50 11.31 33.13 1.40 22.18 1.02 3.05 0.00 0.82 Example 2 0.27 21.76 1.42 11.30 34.52 1.37 21.21 0.70 3.26 0.00 1.16 Example 3 0.30 23.41 1.44 9.55 36.05 1.26 19.26 0.66 3.45 0.00 0.91 Example 4 0.33 24.06 1.44 11.10 32.81 1.20 20.24 0.52 4.14 0.00 0.85 Example 5 0.32 23.66 1.32 9.88 33.27 1.30 18.89 0.52 5.17 0.00 0.55

21 days after dosing 14:00 16:00 16: 1 18: 0 18: 1 18: 1 isomer 18: 2 18: 3 ARA EPA DHA Comparative example 0.22 20.20 1.17 9.38 35.43 1.45 24.13 0.91 2.35 0.00 1.22 Example 1 0.23 20.62 1.52 10.35 33.45 1.37 21.86 0.86 3.28 0.00 1.20 Example 2 0.27 21.70 1.13 10.94 35.00 1.22 20.98 0.72 3.41 0.00 1.11 Example 3 0.26 23.14 1.30 9.92 37.10 1.20 19.60 0.61 3.56 0.00 0.89 Example 4 0.25 23.56 1.62 10.48 33.56 1.20 20.38 0.64 4.36 0.00 0.60 Example 5 0.32 25.18 2.15 9.04 37.16 1.29 16.42 0.39 5.25 0.00 0.49

27 days after dosing 14:00 16:00 16: 1 18: 0 18: 1 18: 1 isomer 18: 2 18: 3 ARA EPA DHA Comparative example 0.18 20.00 1.05 9.17 34.01 1.35 26.52 1.03 2.23 0.00 1.05 Example 1 0.27 20.83 1.19 11.29 34.90 1.36 22.31 0.58 3.25 0.00 1.20 Example 2 0.25 21.92 1.38 10.20 35.37 1.34 21.27 0.80 3.39 0.00 0.99 Example 3 0.26 23.39 2.06 8.65 36.86 1.51 18.97 0.65 3.67 0.00 0.75 Example 4 0.22 22.09 1.50 9.74 35.84 1.44 20.37 0.71 4.33 0.00 0.82 Example 5 0.27 25.44 2.76 8.95 40.26 1.73 12.39 0.31 5.32 0.00 0.50

As can be seen in Table 5 above, after the feed without the dry cells (Comparative Example) and the feed with the dry cells (Example 1 to 5) were administered to the laying hens, the fatty acids in the yolk were analyzed. It can be seen that arachidonic acid is accumulated over%.

As described above, in order to increase the content of arachidonic acid in egg yolk, dry cells obtained by culturing Motierella, a microorganism that produces and accumulate arachidonic acid in cells, are added to laying hens to produce eggs with arachidonic acid accumulated in egg yolk. This became possible. In addition, egg yolk lecithin is produced from eggs and added to the formula to make milk powder close to breast milk.

Claims (6)

Feed composition containing a microorganism producing arachidonic acid. The composition of claim 1, wherein the microorganism is Mortierella alpina. The feed composition of claim 2, wherein the motiella alfina dry cells are added at 0.5 to 15% by weight of the feed. According to any one of claims 2 to 3, wherein the total lipid content of the Motierella alpina dry cells is 20 to 50% by weight, and the content of arachidonic acid in the total lipid is 20 to 50% by weight of the cells. Feed composition. Eggs fortified with arachidonic acid produced by feeding the feed composition of claims 1 to 4 to laying hens. 6. The egg according to claim 5, wherein the content of arachidonic acid in the egg is accumulated in the range of 2 to 10% by weight in egg yolk fatty acid.