KR100381458B1 - Silkworms containing conjugated linoleic acid and rearing process of the insect - Google Patents

Abstract

The present invention relates to a silkworm containing CLA (conjugated linoleic acid), which is an expensive functional oil and a method for preparing the same, comprising the steps of including CLA in mulberry leaves and breeding silkworms with the mulberry leaves. The silkworm containing CLA of the present invention is characterized by containing 1 mg or more of CLA per 1 g of body fat, and in addition to the hypoglycemic effect of the silkworm itself, CLA anticancer, body fat reduction, arteriosclerosis prevention, and immunity enhancement. It has a bioactive effect.

Description

Silkworms containing CLA and a method for producing the same {SILKWORMS CONTAINING CONJUGATED LINOLEIC ACID AND REARING PROCESS OF THE INSECT}

The present invention relates to a silkworm containing CLA (conjugated linoleic acid) which is an expensive functional oil and a method for producing the same. Specifically, a method for accumulating CLA in a silkworm body by raising a mulberry leaf containing CLA and thus It relates to the silkworm containing high content of CLA obtained.

Silkworm ( Bombyx mori L.) has been used as a medicinal herb for a long time. Recently, silkworm powder has been spotlighted as a diabetic agent without side effects as it has been found that silkworm powder has a blood sugar-lowering effect by inhibiting α-glycosidase activity in the human body. . Patents related to "hypoglycemic agents comprising sleep powder as an active ingredient and its manufacturing method" developed by Korean researchers have been registered in Korea and Japan.

On the other hand, CLA (conjugated linoleic acid; conjugated dienoic derivatives of linoleic acid) is the first isomer of linoleic acid isolated and identified from roasted beef. In 1987, it was proved to have strong anticancer properties against skin cancer and breast cancer, and is being classified as a new anticancer substance, and active research is being conducted (Ha, YL Grimm, NK and Pariza, MW, Anticarcinogens from fried ground). beef: heat altered derivatives of linoleic acid.Carcinogenesis, 8, 1881 (1987)). Since it was first synthesized in 1987, CLA has been known to have physiologically active functions such as body fat reduction, arteriosclerosis prevention, and immune function in addition to its anti-cancer effects, and many studies have been conducted. have. These CLAs are found in ruminant meats and dairy products, and beef is known to contain 2.9 to 4.3 mg per g of fat, 2.7 mg for veal and 5.6 mg for lamb.

As such, the amount of CLA is extremely low in foods compared to having various physiological activities, and studies are being made to accumulate it in a large amount in pork, chicken, and fish.

However, no attempt has been made to accumulate CLA in arthropods, such as silkworms, which have no lineage relationship with livestock or fish. Considering the effects of silkworm's hypoglycemia, if blood silk can accumulate large amounts of CLA, In addition to the anti-diabetic effect of lowering, it will be possible to produce a new functional silkworm powder having various effects such as anti-cancer, body fat reduction, arteriosclerosis prevention, immune function enhancement.

In view of the above, an object of the present invention is to obtain a functional silkworm powder that combines the effect of lowering blood sugar of the silkworm and the effects of anticancer, reducing body fat, preventing atherosclerosis, and enhancing immune function of CLA. It is to provide a method for accumulating CLA in high concentrations and silkworms containing CLA obtained in this way.

1 is a graph showing the CLA content accumulated in the body of silkworms bred with mulberry leaves treated with various concentrations of CLA solution,

2 is a graph showing the total fat content accumulated in the body of silkworms bred with mulberry leaves treated with various concentrations of CLA solution,

3 is a graph showing the CLA content accumulated in the body of silkworms bred with mulberry leaves treated with various concentrations of CLA-TG solution,

Figure 4 is a graph showing the total fat content accumulated in the body of silkworms bred with mulberry leaves treated with various concentrations of CLA-TG solution.

Method for producing a silkworm containing the CLA of the present invention for achieving the above object,

Including CLA (conjugated linoleic acid) in the mulberry leaf; And

It characterized in that it comprises the step of breeding silkworms into the mulberry leaves.

Here, the step of including CLA in the mulberry leaves, by applying a solution containing CLA or CLA-TG (conjugated linoleic acid-triglyceride) at a concentration higher than 0.1% (v / v) to the mulberry leaves, CLA per 1 g of mulberry leaves Is preferably 0.1 mg or more, and the solvent is preferably ethanol or acetone.

Silkworm containing CLA of the present invention for achieving the above another object,

It is preferable to contain 1 mg or more of conjugated linoleic acid (CLA) per gram of body fat.

In the present invention, in order to accumulate CLA in the body of silkworms, a method of incorporating CLA into the mulberry leaf, which is a silkworm feed, was used for the silkworm to ingest. At this time, to prepare a mulberry leaf containing CLA, for example, in a solvent such as ethanol or acetone CLA or CLA-TG in a concentration higher than 0.1% (v / v), preferably about 10% (v / v) ) And spray on mulberry leaves. After spraying to the extent that the CLA or CLA-TG solution is completely applied to both sides of the mulberry leaf, the solvent is evaporated to obtain the mulberry leaf coated with CLA or CLA-TG. At this time, the amount of CLA contained in the mulberry leaves is to be 0.1 mg or more per 1 g of mulberry leaves, it is more preferable to be 10 mg or more per 1 g of mulberry leaves. In this way, to obtain mulberry leaves containing CLA, by feeding them from the 3rd age of silkworms and breeding can obtain a silkworm accumulated in a high content of CLA.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these implementations are only illustrations of the present invention, and the present invention is not limited to these.

Example 1

(1) Synthesis of CLA

A CLA having a purity of 90% or more was synthesized from red seed oil. The CLA contains 48% of the two isomers, c9, t11 and t10, c12.

(2) How to accumulate CLA in silkworms

The CLA synthesized above was diluted in concentrations of 0.1, 1, 5, 10 and 15% (v / v) in 99% ethanol, respectively, and evenly sprayed on the mulberry leaves so as to be completely applied on both sides. Next, the mulberry leaves were left in the shade for about 1 hour so as to vaporize the ethanol so that the mulberry leaves were coated with CLA. At this time, when spraying 0.1% CLA solution, at least 0.1 mg of CLA per 1 g of mulberry leaf, at least 1 mg of CLA per 1 g of mulberry leaf in 1% CLA solution, and at least 10 mg of CLA per 1 g of mulberry leaf in 1% CLA solution. It was made. Thus, silkworms were bred by feeding CLA-coated mulberry leaves.

The control group used mulberry leaves sprayed with distilled red seed oil (about 90% linoleic acid content) in ethanol at a concentration of 10% (v / v), and the control group used mulberry leaves that had not been treated. Breeding.

The silkworms wake up from the egg and take four sloughs to make cocoons as 5-year-old silkworms. Until 2, they feed only the mulberry leaves that have not been treated with CLA, and the mulberry leaves that have been treated with CLA. It was raised until the fifth day of the fifth day.

(3) Analysis of CLA content in silkworm body

Silkworms were frozen on day 3 of day 5, and fats were extracted and methylated from them and analyzed by gas chromatography for total fat, CLA content and various other fatty acids.

Statistical significance between each treatment mean was tested at the 5% level according to the general method (Student-Newman-Keuls Test) (SAS Institute, 1989), and there was no significance between treatments indicated by the same alphabet in the figures or tables.

1 is a graph showing the CLA content accumulated in the body of silkworms bred with mulberry leaves treated with various concentrations of CLA solution. Here, it can be seen that the higher the CLA concentration of the CLA solution treated on the mulberry leaves, that is, the greater the amount of CLA contained in the mulberry leaves, the higher the CLA content accumulated in the silkworm body. However, since there is no significant difference between 10% and 15% CLA concentration, the concentration of CLA solution to be treated in mulberry leaves seems to be the most preferable 10%. On the other hand, the CLA solution concentration of 0.1% was detected in a very small amount of CLA, silkworms bred in the mulberry leaves treated with pure mulberry leaves or treated with red seed oil did not have any CLA accumulation in the body.

Figure 2 is a graph showing the total fat content accumulated in the body of silkworms bred in mulberry leaves treated with various concentrations of CLA solution. Here, it can be seen that the total fat content of the silkworm body does not differ depending on the CLA treatment concentration and CLA treatment.

The following Table 1 shows the contents of various fatty acids (mg / fat g) other than CLA accumulated in the body of silkworms bred with mulberry leaves treated with various concentrations of CLA solution.

Processing conditions C14: 0 C16: 0 C18: 0 C18: 1 C18: 2 Γ-LN LN No treatment 3.3a 179.5a 195.8a 229.6b 101.3a 0.6a 289.9a Red seed oil treatment 3.2a 172.2a 186.0a 282.1a 134.7a 0.9a 220.1b CLA 0.1% Treatment 2.4a 140.2b 232.1a 193.7c 131.6a 0.9a 299.2a CLA 1% Treatment 2.2a 138.9b 198.8a 218.7b 119.3a 0.6a 270.8a CLA 5% Processing 2.6a 147.1b 211.4a 275.4a 125.9a 1.2a 235.3b 10% CLA treatment 2.4a 148.1b 205.0a 228.5b 94.6a 1.4a 236.6b CLA 15% Processing 2.5a 150.5b 208.5a 224.3b 108.5a 0.9a 265.8a

In the above table, it can be seen that fatty acid content other than CLA accumulated in the silkworm body does not have a significant difference in the concentration of CLA or with or without CLA treatment.

Example 2

(1) Synthesis of CLA-TG

Since CLA is a free fatty acid, when it is added to animal and vegetable fats and oils, the quality of these fats and oils is feared. To solve this problem, CLA is preferably added to foods in the form of triglyceride derivatives. Neutral fatty acid derivatives of CLA have better absorption in the body than free CLA, and the carboxyl group is removed, thereby eliminating the toxicity caused by the carboxyl group. The preparation method of triglyceride derivatives of CLA (referred to as CLA-TG) in which the hydroxyl group of glycerol is ester-bonded with a carboxyl group at the CLA end is described in Korean Patent Application No. 10-1999-0037151, which is the applicant's prior application.

In the present embodiment, the CLA in the free state synthesized in (1) was synthesized in the form of CLA-TG according to the method of Park et al. (Master's Thesis, Department of Agricultural Chemistry, Gyeongsang National University).

(2) Method of accumulating CLA-TG in silkworm and analyzing CLA content in silkworm

The total fat accumulated in the silkworm body, the CLA content, and various other treatments of silkworms were treated in the same manner as in Example 1, except that CLA-TG was used instead of CLA and acetone was used as a solvent. The content of fatty acids was analyzed.

3 is a graph showing the CLA content accumulated in the body of silkworms bred with mulberry leaves treated with various concentrations of CLA-TG solution. From this, it can be seen that the higher the CLA-TG concentration of the CLA-TG solution treated in mulberry leaves, the higher the content of CLA accumulated in the silkworm body. There was no significant difference between 10% and 15% CLA-TG concentration. On the other hand, the CLA-TG solution concentration of 0.1% was detected in a very small amount of CLA, CLA-TG did not have any CLA accumulation in the silkworms were bred with mulberry leaves treated with pure mulberry leaves or red seed oil. Comparing this result with FIG. 1, it can be seen that the content of CLA accumulated in the body tends to be slightly higher in silkworms grown with mulberry leaves treated with CLA-TG than in the case of treatment with free CLA.

Figure 4 is a graph showing the total fat content accumulated in the body of silkworms bred in mulberry leaves treated with various concentrations of CLA-TG solution. Here, it can be seen that the total fat content in the silkworm body was not different depending on the concentration of CLA-TG and CLA-TG. Compared with FIG. 2, it can be seen that the amount of total fat accumulated in the body is slightly higher in silkworms bred with mulberry leaves treated with CLA-TG than in the case of treatment with free CLA.

Table 2 shows the contents of various fatty acids (mg / fat g) other than CLA accumulated in the body of silkworms bred with mulberry leaves treated with various concentrations of CLA-TG solution.

Processing conditions C14: 0 C16: 0 C18: 0 C18: 1 C18: 2 Γ-LN LN No treatment 3.1a 190.0a 198.0a 210.2a 115.2a 0.8a 227.9b Red seed oil treatment 3.9a 185.3a 186.2a 230.1a 145.6a 0.7a 238.1b CLA-TG 0.1% Treatment 3.2a 146.8b 190.5a 199.7b 132.8a 1.5a 246.7b CLA-TG 1% Treatment 3.0a 179.2a 190.2a 222.9a 108.2a 1.2a 222.3b CLA-TG 5% Treatment 3.7a 160.3b 206.6a 240.6a 120.4a 1.5a 266.2a CLA-TG 10% Processing 3.4a 152.3b 205.8a 219.8a 129.5a 1.8a 242.9b CLA-TG 15% Processing 3.9a 168.8b 215.5a 239.9a 109.6a 1.9a 291.3a

In the above table, it can be seen that fatty acid content other than CLA accumulated in the silkworm body does not have a significant difference in the concentration of CLA-TG or depending on the presence or absence of CLA-TG treatment.

As described above, when CLA or CLA-TG is fed to a mulberry leaf sprayed by diluting the appropriate concentration in a solvent such as ethanol or acetone from the 3rd stage of silkworm, the CLA is high in the silkworm body. It can be seen that the accumulation.

According to the present invention, according to the method of breeding silkworms with mulberry leaves containing CLA, silkworms containing a high content of CLA can be obtained. In addition, physiological activities such as reducing body fat, preventing atherosclerosis and enhancing immunity can be expected.

Claims (6)

Including CLA (conjugated linoleic acid) in the mulberry leaf; And Characterized in that it comprises the step of breeding silkworms with the mulberry leaves. The method of claim 1, wherein the method comprises at least 0.1 mg of CLA per gram of mulberry leaf. The method of claim 1 or 2, wherein the step of including CLA in the mulberry leaf, applying a solution containing CLA or CLA-TG (conjugated linoleic acid-triglyceride) at a concentration higher than 0.1% (v / v) to the mulberry leaf How to do. The method of claim 3 wherein the solution containing CLA or CLA-TG is a solvent of ethanol or acetone. Silkworm, characterized by containing CLA (conjugated linoleic acid). The silkworm of claim 5, which contains at least 1 mg of CLA per gram of body fat.