KR20100138495A - Process for producing lutein-enriched eggs with feed additive comprising fermented mandarin peel - Google Patents

Abstract

PURPOSE: A producing method of lutein enriched egg, using a feed additive containing fermented mandarin peel is provided to improve the content of lutein inside eggs. CONSTITUTION: A producing method of lutein enriched egg comprises the following steps: adding 3~10parts of ground mandarin peel by weight, 0.5~2parts of soybean powder by weight, and 0.03~0.1parts of dipotassium phosphate by weight with 100parts of water; controlling the ph of the mixture into 6.5~7.2; injecting bacillus subtilis MORI for fermenting in 35~37deg c; condensing the fermented liquid under the decompression condition, and hot-air drying to make fermented mandarin peel powder; mixing the fermented mandarin peel powder with an excipient to make a feed additive; and mixing the feed additive with feed for chicken.

Description

PROCESS FOR PRODUCING LUTEIN-ENRICHED EGGS WITH FEED ADDITIVE COMPRISING FERMENTED MANDARIN PEEL}

The present invention relates to a method for producing lutein fortified eggs using a feed additive containing tangerine peel fermentation, and more particularly, fermentation of lutein-containing tangerine peel using Bacillus subtilis MORI strain. By preparing a tangerine peel fermented product with improved lutein absorption, and preparing a feed additive including the tangerine peel fermented product and adding the same to a laying hen's feed to feed the laying hens, thereby producing an egg with enhanced lutein content in the egg; It relates to lutein-enriched eggs produced by the method.

Carotene-like pigment group, a carotinoid widely distributed in animal and plant systems, can be extracted by adding organic solvents such as acetone to fresh tissue.It is an unstable substance that is easily oxidized in air and is insoluble in water. It is divided into carotene which does not contain oxygen and xanthophylls which contain oxygen. Carotenoids have been reported to prevent the oxidation of DNA or saturated fatty acids, the major substances in the body by removing harmful oxygen from the body (Holt S. Soya, Korean Soybean Digest. 14: 77-90, 1977).

Carotenoids are composed of eight isoprenoids and are essential in vivo, derived from the structure of C ₄₀ H ₅₆ and acting as strong antioxidants. Free radicals enter the living body from living areas such as aromatic hydrocarbons, tobacco, pesticides, organic solvents, fried foods, alcohol, and air pollution, and combine with proteins and unsaturated fats. Destroys DNA, RNA, etc., and weakens immunity and causes various diseases, wrinkles and ages. Carotenoids play an important role in slowing this rate of oxidation, preventing the oxidation of low-density lipoproteins, preventing heart disease due to coronary blood flow disorders, and aryl hydrocarbon hydroxylases (aryl). It also contributes to the detoxification of carcinogens by altering the activity of hydrocarbon hydroxylases [Rom. J. Intern. Med., 35, 55-62, 1997].

In addition, carotenoids, such as retinoids and vitamin D (Vit D), have been reported to be practical substances for skin cancer. Investig. Dermatol. Symp. Proc., 1, 15-21, 1996]. Carotenoids such as lycopen, beta-carotene, alpha-carotene, lutein, canthaxanthin, and astaxanthin may have been mixed rather than alone. It has been reported to have a synergistic effect, and when lutein or lycopene are present together, the effect becomes greater and thus plays an important role in preventing aging and human diseases. [FEBS Lett., 427, 305- 308, 1998].

Lutein, a carotenoid pigment, is excellent in antioxidant function and has proven to be effective in preventing eye diseases such as age-related macular degeneration and cataracts, heart disease, skin cancer and breast cancer. Lutein is an isomer of zeaxanthin, which is mainly present in the intraocular tissues, i.e., the macula of the lens and retina, and dark leafy vegetables such as spinach and broccoli are known as the main source of food (Bone RA, Landrum JT, Tarsis SL. Res. 24: 1521-1535, 1985).

Lutein is a pigment present in the largest amount of xanthophylls of the carotenoid represented by the formula C ₄₀ H ₅₆ O ₂ , which is also called xanthophyll.

Recent studies have shown a decreased incidence of cataracts in people with high blood carotenoid concentrations and lutein intake may reduce the risk of vision impairment in the elderly population (Chasab-Taber. Et al, Am. J. Clin). Nutr. 70: 509-516, 1999) (Seddon JM. Et al, J. AM. Med. Assoc. 272: 1413-1420, 1994).

In addition to the function of vision protection, lutein has been reported to have immunity and anti-cancer activity and to be correlated with the prevention of various chronic diseases such as preventing skin damage and preventing cardiovascular diseases (Granado F. et al, Brit. J. Nutr). 90: 487-502, 2003 (Mares-Perlman JA. Et al, Overview.J. Nutr. 132: 518S-524S, 2002).

Tangerine, a representative fruit in winter in Korea, contains a large amount of various carotenoids, including lutein, in its skin, and these carotenoids have been reported to affect anticancer, immunity, and heart disease (Whang, HJ & Yoon, KR, Kor. J. Food Sci.Technol. 27 (6): 950-957, 1995). In addition, bioflavonids have antioxidant, antibacterial, and anticancer properties based on diphenylpropane (Brack, M..E., Et al., Food Technol. 48: 121-124, 1994 Naringin and hesperdin are known to have antibacterial and blood pressure lowering effects (Han, SS & You, IJ, Kor. J. Mycol. 16:33). -40, 19878: Son, HS et al., J. Korean Soc.Food Sci.Nutr., 21: 136-142, 1992).

Therefore, the present inventors found a method of increasing the extraction yield of carotenoids and lutein from tangerine peel by fermenting the tangerine peel using microorganisms and finding a method of producing eggs having excellent lutein content by feeding lutein-containing feed using the same. In the meantime, the Bacillus subtilis MORI strain (hereinafter referred to as the 'MORI strain'), which is suitable for increasing lutein production, was selected, and the feed used to improve lutein absorption was produced using the MORI strain. The present invention was completed by confirming that lutein content is increased in eggs produced from laying hens.

Therefore, an object of the present invention is a lutein having an improved lutein content in eggs produced by mixing feed additives, including a tangerine peel fermented product prepared by fermentation with Bacillus subtilis MORI strain, to feed the laying hens. It is to provide a method of producing fortified eggs.

Another object of the present invention is to provide a lutein fortified egg with an increased lutein content obtained by the above method.

Hereinafter, the present invention will be described in detail.

3 to 10 parts by weight of dried tangerine pulverized powder, 0.5 to 2 parts by weight of soybean meal and 0.03 to 0.1 parts by weight of dibasic potassium phosphate were mixed, and the pH of the mixture was adjusted to 6.5 to 7.2 with 5N sodium hydroxide. Adjusting;

Bacillus subtilis MORI ( Bacillus subtilis MORI, KCCM-10450) inoculated at a concentration of 1 to 10 w / v% in the total culture solution to ferment at 35 to 37 ℃;

Concentrating the fermentation broth obtained above under reduced pressure and hot air drying to prepare a tangerine peel fermented product in powder form, and mixing the tangerine peel fermented product with an excipient to prepare a feed additive; And

 Feed additives containing the dried tangerine peel fermented product is mixed with feed for laying hens to feed the laying hens and obtaining eggs from the laying hens, characterized in that the feed additive containing a tangerine peel fermentation It is a method of producing lutein fortified eggs using.

Hereinafter, the present invention will be described in more detail.

The present invention, 1) grinding the tangerine peel; 2) fermenting the pulverized tangerine peel by inoculating Bacillus subtilis MORI (Accession No .: KCCM-10450) strain; 3) concentrating and drying the fermented tangerine peel mill; 4) preparing a feed additive by mixing the dried tangerine peel fermented product with an excipient; 5) preparing a feed for laying hens by adding a feed additive comprising a tangerine peel fermented product to the feed; And 6) a method of producing a lutein-enriched feed and a lutein-enriched egg, which includes feeding the feed containing the tangerine peel fermentation additive to the laying hen.

In the present invention, the tangerine peel in step 1) may be used by natural drying or hot air drying, and then pulverized to 100 mesh or less, and pulverized into crude powder of less than that.

3 to 10 parts by weight of the obtained orange peel ground powder, 0.5 to 2 parts by weight of soy flour or defatted soy flour, and 0.03 to 0.1 parts by weight of dibasic potassium phosphate were mixed with 100 parts by weight of water, followed by 5 N sodium hydroxide (NaOH). The pH is adjusted in the range of 6.5 to 7.2. The nutrient source for microbial growth provides optimum growth conditions for microorganisms in the next fermentation step by adjusting the pH of the mixture comprising soy flour or skim soy flour and tangerine husk pulverized with dibasic potassium phosphate. Sterilization of this mixed solution may be carried out by a conventional method, for example, 121 ° C., for 20 minutes.

The mixture including the tangerine peel may be fermented after inoculation by inoculation of the microorganism in an amount of 1 to 10 w / v%, preferably 2 to 8 w / v%, based on the total culture volume.

A microorganism which is added to the fermentation of orange peel in the above step 2) is Bacillus subtilis, Bacillus licheniformis, Bacillus polyfermenticus such as Bacillus (Bacillus sp.), Microbial and feed added probiotics in Lactobacillus bacteria of microorganisms that can be used as for (Lactobacillus sp. ), Yeast ( Saccharomyces cerevisiae ) and the like, preferably Bacillus subtilis MORI (Accession Number: KCCM-10450) is used.

The Bacillus subtilis MORI strain used in the present invention is a microorganism similar to the bacterium used for soybean fermentation, producing polygammaglutamic acid, a biological surfactant, and the like, which promote the absorption of useful substances in the intestinal tract. In addition, the fermentation metabolites of these strains are safe and contain various other functional ingredients, which is very beneficial when consumed by humans.

The fermentation of the microorganism in the above is fermented at a temperature of 30 to 40 ℃, preferably 35 to 37 ℃, 2 to 5 days, preferably 3 to 4 days at pH 4 to 9, preferably pH 6 to 7 .

In the present invention, it is possible to provide a feed comprising a tangerine peel fermented product improved lutein absorption prepared by the above method.

The tangerine peel fermented product obtained in step 2) is concentrated in vacuo and then, according to a conventional method, in the form of a conventional formulation, for example, a liquid form, a water-soluble powder, an emulsifiable concentrate, a tablet, a suspension, a powder, and a water-dispersible granule. It can be formulated as. According to the present invention, feed by mixing at least one excipient selected from the group consisting of marigold extract, wheat bran, rice bran, ethanol, soybean meal, protein skin, zeolite, bentonite, vermiculite and elvanite with the tangerine peel fermentation Additives can be prepared.

Formulated feed additives obtained by the method of the present invention may be added to feed for all kinds of animals raised in farms, but preferably added to feed of cattle, pigs or poultry for laying meat and laying hens for obtaining eggs. can do.

The mixture containing the tangerine peel fermented product is significantly improved the absorption of lutein than the untreated fermented tangerine peel when added to the feed for laying hens as a feed additive can increase the lutein content in eggs obtained from laying hens fed with this feed have.

Figure 1 is a yolk egg yolk (c), and tangerine peel fermented with feed supplemented with 0.1% by weight of a tangerine peel fermentation concentrate (a), control (no treatment, b), a feed additive containing a tangerine peel fermentation The egg yolk (d), which was fed by feeding the feed containing 0.2 wt% of the feed additive including water, was extracted from lutein by the method of Example 4, and the chromaticity was compared. As shown in Figure 1, the control (b) is significantly lower enough to be seen with the naked eye than the color (c) and (d) of egg yolks laid by feeding the feed containing the tangerine peel fermented products according to the present invention It can be seen that the lutein content in egg yolk obtained by the method is higher.

Tangerine peel fermented product obtained by the method of the present invention may be included in an amount of 2 to 100% by weight, preferably 5 to 40% by weight based on 100% by weight of the total feed additive. Feed additives containing the tangerine husk fermented product obtained in accordance with the method of the present invention may be added in the feed for laying hens 0.1 to 0.3% by weight relative to 100% by weight of the whole laying hens, preferably 0.2% by weight. .

According to the method of the present invention, through the tangerine peel fermentation by microorganisms can produce a large amount of tangerine peel fermented product with improved lutein absorption, and the feed additive is prepared by mixing the tangerine peel fermented product obtained by the above method with an excipient And, by adding this to the feed for laying hens breeding eggs and obtaining eggs can be produced lutein fortified eggs with improved lutein content than conventional eggs.

By feeding the laying hens containing the tangerine bark fermented product produced according to the method of the present invention to the laying hens, the lutein absorption is significantly improved compared to the existing laying hens, thereby preventing, improving and treating eye diseases such as macular degeneration and cataracts. It can be usefully used, and can produce an egg as a functional food capable of improving anticancer function such as heart disease improvement function, skin cancer, breast cancer and the like.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to illustrate the present invention, but the scope of the present invention is not limited thereto.

Example 1 Preparation of Tangerine Peel Ferment

Dried tangerine peel was purchased at Gyeongdong Market, Seoul, and used as a powder to pulverize to 100 mesh or less.

In a 500 L fermentor, 300 kg of water, 15 kg of tangerine husk, 3 kg of soy flour, 0.3 kg of dibasic potassium phosphate, and the like were mixed with water to adjust the pH to 7.0 with 5 N NaOH. For sterilization, Bacillus subtilis MORI spawn prepared by incubation at 121 ° C. for 20 minutes, cooled, and then cultured in YM medium (BD, France) was inoculated in an amount of 2 w / v% based on the total fermentation broth volume. After fermentation temperature of 37 ℃, fermentation time 3 days, the fermentation broth was concentrated to 30 L under reduced pressure at 60 ℃ and dried by hot air to prepare a tangerine peel fermented product in powder form.

The MORI strain having the nucleotide sequence of SEQ ID NO. 1 shown in FIG. 2 was named " Bacillus subtilis MORI MORI" and deposited on November 26, 2002 at the Korea Microorganism Conservation Center No. KCCM-10450. Deposited in an arc.

To 15 kg of tangerine peeled fermented product obtained above, 10 kg of soybean meal, 10 kg of rice bran and 500 g of marigold extract were mixed with a V-mixer (product of Korea, Korea) as an excipient to prepare a feed additive.

Example 2 Preparation of a Feed Containing Tangerine Peel Fermentation

Feed additives, including the tangerine peel fermented product prepared in Example 1, 0 kg (0.0% by weight, control), 1 kg (0.1% by weight), respectively, in the feed composition for laying hens according to the ingredient table described in Table 1 below, And 2 kg (0.2% by weight) was mixed to make a total feed amount of 1000 kg to prepare a feed for laying hens containing tangerine peel fermentation.

Raw material name content(%) corn 65.0 Lupine seeds 5.0 Soybean meal 15.0 Six minutes 3.0 Limestone 10.0 Dicalcium Phosphate 0.5 Salt 0.3 Uji 0.8 Choline chloride (50%) 0.1 DL-Methionine 0.1 Mineral mix 0.1 Vitamin mix 0.1 Sum 100

Example 3 Spectrometer Specification Test

30-week-old Hy-Line Brown laying hens were laid in a cage for laying hens so that their weight and egg weight were similar to each other, and then the feed for the laying hens was mixed with a feed additive comprising a tangerine peel fermented product prepared in Example 2. Was used for the experiment.

All three treatment groups consisted of 10 water for each test group, and three test groups were provided to the laying hens, such as a test group fed a feed containing 0.1 wt% and 0.2 wt% of an untreated control and a tangerine peel fermented product. A pay test was conducted for four weeks. Eggs were collected after feeding and analyzed for lutein content.

Example 4 Analysis of Lutein Components in Tangerine Peel Fermentation and Eggs

Changes in the lutein content of tangerine peeled and tangerine peeled fermentation were compared, and the changes in lutein content in each egg collected from control diet fed laying hens and diets with improved lutein absorption were measured.

Take 2 g of each sample of tangerine peel and tangerine peel fermentation and dissolve in 8 ml ethanol, and for each egg, filter it using 710 micrometer mesh to separate egg yolk, take 2 g of each egg yolk and dissolve in 8 ml of ethanol. I was.

Then, the extract was shaken at 90 ° C. at 4 ° C. and 160 rpm. After centrifugation at 3000 rpm for 5 minutes, the supernatant was placed in a 50 ml volumetric flask. 10 ml of tetrahydrofuran was added to the precipitate, mixed thoroughly, and centrifuged at 3000 rpm for 5 minutes to transfer the supernatant to a 50 ml volumetric flask. This process was repeated four times.

The final amount was adjusted to 50 ml and the supernatant was taken by centrifugation at 3000 rpm for 5 minutes. 1 ml of the supernatant was taken and dried using nitrogen gas. 1 ml of ethanol was added to the dried supernatant and dissolved, and then measured by spectrophotometer at 445 nm.

The lutein content was calculated in ppm by applying the following formula, and the results are shown in Table 2 below.

The results are expressed as mean and standard deviation (mean ± standard deviation). The significance of lutein content between test groups was verified at P <0.05 using Duncan's multiple test.

As a result, the lutein content of the tangerine peel fermented by Bacillus subtilis MORI was measured to be 200.30 ppm compared to the lutein content of the tangerine peel dried before the fermentation was 80.86 ppm. It was confirmed that the lutein content in water increased by more than 2.4 times.

In addition, the lutein content in the untreated control egg was 26.95 ppm, whereas the egg fed the feed containing 0.1 wt% of the feed additive containing tangerine peel fermented was 34.57 ppm and the egg containing the feed containing 0.2 wt%. In case of lutein content of 42.00 ppm.

Therefore, in the case of chickens fed a feed containing tangerine peel fermented products, a feed containing 0.1 wt% tangerine peel fermented feed additive containing 1.3% and 0.2% by weight compared to the control untreated eggs Feeding with feed additives containing tangerine peel fermented products showed significantly higher lutein content ( P <0.05).

Sample Name Lutein (ppm) Duncan Multiple Statistics Analysis Tangerine peel 80.9 ± 0.9 Tangerine Peel Ferment 200.3 ± 3.9 Control (no treatment) 27.0 ± 1.0 c Feed feed with 0.1 wt% tangerine peel fermented feed additive 34.6 ± 0.8 b Feeding feed with 0.2% by weight tangerine peel fermented feed additive 55.0 ± 0.8 a

In Table 2, a, b, and c are Duncan's grouping, and as a result of the SAS program, a, b, and c represent different groups, thereby proving that each group is significant. For example, if each group is formed as a, a, a means that the groups are the same as each other, it means that each group is not significant to each other.In another example, when a group is formed as a, ab abc, a It means that each group is not significant with each other. Therefore, the results indicate that each group was formed differently as a, b, and c at the significance level P <0.05, indicating a significant result.

The present invention is not limited to the technical spirit of the specific examples, test examples, drawings or tables described above, and the general knowledge in the technical field to which the invention belongs without departing from the spirit of the invention claimed in the claims. Of course, any person having a variety of modifications can be made, and such changes are within the scope of the claims.

Figure 1 is a yolk egg yolk (c), and tangerine peeled by feeding the feed added 0.1% by weight of the tangerine peel fermentation concentrate (a), control (no treatment, b), feed additive containing a tangerine peel fermentation The feed is added to the feed additive containing 0.2% by weight of the fermented egg yolk (d) of the egg was laid lutein extracted by the method of Example 4 to compare the chromaticity.

Figure 2 shows the 16S rDNA sequence of the Bacillus subtilis MORI strain used in the present invention.

Claims (5)

3 to 10 parts by weight of dried tangerine pulverized powder, 0.5 to 2 parts by weight of soybean meal and 0.03 to 0.1 parts by weight of dibasic potassium phosphate were mixed, and the pH of the mixture was adjusted to 6.5 to 7.2 with 5N sodium hydroxide. Adjusting; Bacillus subtilis MORI ( Bacillus subtilis MORI, KCCM-10450) inoculated at a concentration of 1 to 10 w / v% in the total culture solution to ferment at 35 to 37 ℃; Concentrating the fermentation broth obtained above under reduced pressure and hot air drying to prepare a tangerine peel fermented product in powder form, and mixing the tangerine peel fermented product with an excipient to prepare a feed additive; And  Feed additives containing the dried tangerine peel fermented product is mixed with feed for laying hens to feed the laying hens and obtaining eggs from the laying hens, characterized in that the feed additive containing a tangerine peel fermentation Method of producing lutein fortified eggs using. The method of claim 1, wherein the Bacillus subtilis MORI strain is inoculated 2 to 3 w / v% based on the total culture volume production of lutein fortified eggs using a feed additive containing tangerine peel fermentation Way. The tangerine peel according to claim 1, wherein the excipient is at least one component selected from the group consisting of marigold extract, bran, rice bran, brew, soybean meal, protein peel, zeolite, bentonite, vermiculite and elvan. Method for producing lutein fortified eggs using a feed additive containing fermented products. The lutein fortified egg using the feed additive containing tangerine peel fermentation according to claim 1, wherein the feed additive comprising the tangerine peel fermentation is added in an amount of 0.1 to 0.3% by weight based on 100% by weight of the whole feed. Production method. Lutein fortified eggs, which are produced by the method according to any one of claims 1 to 4.

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