KR20030062651A - A Media of Monascus.sp. for feed additives and its method and products - Google Patents

Abstract

PURPOSE: A medium of Monascus sp. for feed additives, a production method thereof and the use of products produced thereby are provided, thereby cheaply producing high concentration of second metabolites of Monascus sp. having cholesterol lowering effects. CONSTITUTION: A medium of Monascus sp. for feed additives comprises (a) proflo which is an extract of cotton seed as a nitrogen source; (b) one material selected from the group consisting of sugar, rice powder, corn powder, potato powder, wheat flour and the mixture thereof; and (c) Na, Ca, Fe, Cu and Mn as trace elements. A method for producing the second metabolites of Monascus sp. comprises aerobically culturing Monascus sp. in the medium of Monascus sp. for feed additives for 6 to 8 days.

Description

Hongkuk culture medium for livestock feed additives, methods and products {A Media of Monascus.sp. for feed additives and its method and products}

The present invention relates to a hongguk culture medium for livestock feed additives, and a method and a product thereof. In particular, the present invention maximizes the production of secondary metabolites of hongguk bacteria having a cholesterol lowering effect and at the same time the liquid culture medium and cultivation method of hongguk bacteria using the same, the culture medium of hongguk bacteria obtained therefrom It is about.

Microorganisms of the genus Monascus called red yeast bacteria have a characteristic of secreting a dark red pigment and a cholesterol biosynthesis inhibitor called monacolin K, which has been used by humans for various purposes for hundreds of years. On the food side, red yeast rice is used in food preservatives and red colorants, and red dye in the dyeing industry. In addition to food and dyeing engineering, the most useful fields of red yeast bacteria are human foods made by fermenting red yeast yeast, red yeast fermented rice, and red yeast fermented bread. They are used as a type of cholesterol inhibitor to lower human blood cholesterol levels.

Hongguk has long been known to be effective in lowering blood cholesterol levels in humans, as it was recorded for blood circulation purposes in the Chinese Ming Dynasty, Lee Si-jin. Since fermented foods of Hongkuk bacterium have a blood sugar lowering effect and blood pressure lowering effect in addition to the purpose of lowering blood cholesterol, they are being used as a diabetic treatment aid and a blood pressure lowering aid.

As described above, red yeast bacteria have been widely developed as cholesterol-lowering drugs and cholesterol-lowering functional foods, but have not been used for livestock feed. In order to obtain low cholesterol animal food using red yeast fungi as a livestock feed, it is necessary to effectively cultivate red yeast fungi at a very low price due to the characteristics of livestock feed. Currently, the method of culturing red yeast bacteria is a method of cultivating a solid culture method and a grain-derived grain derived from the grains as a main ingredient and adding a few minerals. Table 1 shows representative erythrocyte solid culture methods known to date. The problem with this solid culture method is that red yeast bacteria breed well in rice, but low propagation rate in soybeans, corn, wheat and other grains. Pollution by various germs has emerged as the most critical problem. In order to solve the problems of the solid culture method has been developed a liquid culture method has been able to cultivate the active ingredient hongguk at high concentration without worrying about contamination of various bacteria. Table 2 shows representative hongkuk liquid culture methods known to date. However, the liquid culture method has a problem that there is no big problem in the development of medicines and functional foods for humans due to the high cost of culturing red yeast bacteria, but it is not economical at all to develop them as livestock feed.

Recently, a method of preparing chicken feed (Korean Patent Application No. 10-2001-0028676) for producing low cholesterol eggs and broilers by culturing red yeast bacteria in soybeans by applying the general method of the solid culture method has been developed. However, this method is very inexpensive to raw materials, but there is a serious cost in the medium selection and dehydration process, sterilization process and drying process, and there is a problem in commercialization due to the contamination problem by various bacteria as in other solid culture methods.

Solid culture medium badge Ingredient Remarks Ariel 20g rice (soaked with 0.015M ZnSO ₄ ) Ohantaek 120mg rice (soaked with 12ml of 0.128M ZnSO ₄ , 68ml H ₂ O) Stuart 40g rice (soaked with 20ml Distilled Water) Lin _{30 rice powder, 1.5g NaNO 3,} MgSO 4 · 7H 2 O, 2.5g KH 2 PO 4

Liquid culture medium badge Ingredient (per liter) Remarks SP 100 g sucrose, 10 g peptone, 1 g KH ₂ PO ₄ , 0.5 g MgSO ₄ 7H ₂ O, 1 g Tartaric acid, 3 g L-aspartic acid YM 3g yeast extract, 3g malt extract, 5g peptone, 10g glucose YE 40g glucose, 3g KH ₂ PO ₄ , 10g yeast extract GMin 40 g glucose, 1 g KH ₂ PO ₄ , 0.5 g MgSO ₄ 7H ₂ O, 0.5 g NaCl 0.1 g FeSO ₄ 7H ₂ O SMO 100 g sucrose, 1 g peptone, 2 g KNO ₃ , 3 g tartaric acid (0,2 g (NH ₄ ) ₂ HPO ₄ , 0.5 g MgSO ₄ 7H ₂ O, 0.5 g ZnSO ₄ 7H ₂ O, 0.1 g CaCl ₂ 2H ₂ O MY 20g malt extract, 2g yeast extract GY 100 g glucose, 8 g yeast extract Nishikawa 100g of sucrose, 10g of _{peptone, 2g (NH 4) 2 HPO} 4, 2g KNO 3 5g MgSO 4 · 7H 2 O, 0.1g ZnSO 4 · 7H 2 O, 0.1g CaCl 2 · 2H 2 O, 3g tartaric acid, 0.5g AS ₂ 0 ₃ Sucrosemedium 100 g glucose, 3 g yeast extract, 5 g casamino acid, 2 g NaNO ₃ , 1 g KH ₂ PO ₄ , 0.5 g MgSO ₄ 7H ₂ O, 0.5KCL, 0.01G FeSO ₄ Mizutani 50 g glucose, 20 g peptone 8 g KH ₂ PO ₄ , 0.5 g MgSO ₄ 7H ₂ O, 2 g CH ₃ COOK, 1 g NaCL Toya 30 g glucose, 1.5 g NaNO₃, 1 g MgSO _{4 7} H ₂ O, 2.5 g KH ₂ PO ₄

The present invention relates to a hongguk liquid culture method that has solved the problem that could not be used as a feed additive for livestock due to economic problems, although the liquid culture method of hongguk bacteria has various advantages, in the present invention, secondary metabolic process in a short time at a low cost Provided is a liquid culture medium capable of culturing this highly activated erythrocytes and a culture method using the same.

When the cultured cell culture solution of the honggukyun cultured as a feed additive is fed to the livestock, the cholesterol level of the livestock can be lowered, and various low-cholesterol animal foods can be obtained from the livestock.

Therefore, another aspect of the present invention relates to the use of the cell culture medium of hongguk bacteria cultured by the liquid culture method as described above, livestock feed additive comprising the cell culture medium of the present invention as an active ingredient and a method of producing low-cholesterol livestock using the same And animal foods such as low cholesterol meat products, eggs and milk obtained therefrom.

In addition, the present invention provides a functional food and hyperlipidemia therapeutic agent using the cell culture medium of the present invention.

Figure 1 shows the cholesterol content of the eggs obtained after the 21 days of breeding by adding the Hong Kong mycelium culture medium of the present invention cultured using a PST production medium to chicken feed.

Figure 2 shows the cholesterol content of the eggs obtained after the 21 days of breeding by adding the Hong Kong mycelium culture medium of the present invention cultured using a PRT production medium to chicken feed.

As used herein, the term "cell culture medium" is used to include both cells grown in the medium and products produced therefrom, in particular, secondary metabolites and culture medium.

As used herein, "hongguk" means fungal microorganisms of the genus Monascus that secrete monacolin K.

As used herein, the term "livestock" is used to include poultry such as chickens, ducks, ostriches, and the like, and animals such as cattle, pigs, goats, sheep, and other animals raised on farms or homes for food.

Culture medium of red yeast bacteria for activation of secondary metabolic process and increase of bacterial density

Hongkuk culture medium of the present invention, in the liquid culture medium of Hongguk bacteria consisting of nitrogen, carbon source, microelements essential for the growth of bacteria, for the activation of secondary metabolic process and increase of the density of the fungus (a) cotton seed extract as a nitrogen source pro Proflo;

(b) a material selected from the group consisting of sugar, rice flour, corn flour, potato flour, flour and mixtures thereof as a carbon source; (c) Sodium (Na), calcium (Ca), iron (Fe), copper (Cu) and manganese (Mn) components as trace elements.

Preferably, the erythrocyte culture medium is 0.5 to 1.5% proplo, 1.5 to 4% carbon source, 0.1 to 0.5% NaCl, 0.1 to 0.5% CaCO₃, 0.01 to 0.04% FeCl ₃ H ₂ O, CuCl ₂ · 2H ₂ O 0.001 to 0.002%, MnCl ₂ · 4H ₂ O 0.001 to 0.002%, ZnCl ₂ 0.002 to 0.006%, Na ₂ B ₄ O ₇ · 10H ₂ O 0.001 to 0.002%, (NH ₄ ) ₆ Mo ₇ O ₂₄ 0.001 to 0.002% of 4H ₂ O and the balance of distilled water.

Specific examples of the honggukyun liquid culture medium according to the present invention are shown in Table 3 below. However, the red yeast cell culture medium of the present invention is not limited thereto.

Badge Name Ingredient (per liter) PST 10 g proflo, 22.5 g sucrose, 3 g NaCl, 3 g CaCO₃, 40 mg ZnCl ₂ , 200 mg FeCl ₃ · 6H ₂ O, 10 mg CuCl ₂ · 2H ₂ O, 10 mg MnCl ₂ · 4H ₂ O, 10 mg Na ₂ B ₄ O ₇ 10 H ₂ O, 10 mg (NH ₄ ) ₆ Mo ₇ O ₂₄ 4 H ₂ O PRT 10g proplo, 22.5g rice powder, 3g NaCl, 3g CaCO₃, 40mg ZnCl ₂ , 200mg FeCl ₃ · 6H ₂ O, 10mg CuCl ₂ · 2H ₂ O, 10mg MnCl ₂ · 4H ₂ O, 10mg Na ₂ B ₄ O ₇ 10 H ₂ O, 10 mg (NH ₄ ) ₆ Mo ₇ O ₂₄ 4 H ₂ O PCT 10 g proplo, 22.5 g corn powder, 3 g NaCl, 3 g CaCO₃, 40 mg ZnCl ₂ , 200 mg FeCl ₃ · 6H ₂ O, 10mg CuCl ₂ · 2H ₂ O, 10mg MnCl ₂ · 4H ₂ O, 10mg Na ₂ B ₄ O ₇ 10 H ₂ O, 10 mg (NH ₄ ) ₆ Mo ₇ O ₂₄ 4 H ₂ O PPT 10 g proplo, 22.5 g potato powder, 3 g NaCl, 3 g CaCO₃, 40 mg ZnCl ₂ , 200 mg FeCl ₃ · 6H ₂ O, 10 mg CuCl ₂ · 2H ₂ O, 10 mg MnCl ₂ · 4H ₂ O, 10 mg Na ₂ B ₄ O ₇ 10 H ₂ O, 10 mg (NH ₄ ) ₆ Mo ₇ O ₂₄ 4 H ₂ O PWT 10 g proplo, 22.5 g wheat flour, 3 g NaCl, 3 g CaCO₃, 40 mg ZnCl ₂ , 200 mg FeCl ₃ · 6H ₂ O, 10mg CuCl ₂ · 2H ₂ O, 10mg MnCl ₂ · 4H ₂ O, 10mg Na ₂ B ₄ O ₇ 10 H ₂ O, 10 mg (NH ₄ ) ₆ Mo ₇ O ₂₄ 4 H ₂ O

By culturing hongguk bacteria using the culture medium of the present invention as described above can obtain a cell culture medium containing a large amount of cholesterol-lowering substances such as a large amount of secondary metabolites, that is, monacholine K. Hereinafter, a method of culturing hongguk bacteria using the hongguk culture medium according to the present invention will be described in detail. The culturing method of hongguk bacteria according to the present invention is characterized in that it is cultured in the liquid culture medium of the present invention, the present invention is not limited by the detailed liquid culture method described below.

Cultivation method of Hongguk bacteria with maximum secondary metabolite and bacteria density

Some fungi have a secondary metabolic process in addition to the primary metabolic process, such as Hong Kuk-gyun. Primary metabolism is a metabolic process common to all living organisms. Secondary metabolism is not found in animals and is found in some microorganisms and plants. To produce chemicals. In the case of red yeast bacteria, cells grow and divide through the first metabolic process, and through the second metabolic process, monacoline K, monascin, ankaflavin, rubropunctatin, and monascorbulin (monascorubrin), rubropunctamine, monascorubramine and various chemicals are secreted. Chemicals such as Monacoline K inhibit the animal's cholesterol biosynthesis process, which lowers the body's cholesterol levels.

To use red yeast bacteria as a livestock feed, you can mix them with a feed as a feed additive. At this time, in order to effectively lower the cholesterol level of livestock, the secondary metabolic process of red yeast bacteria should be fully expressed. Since erythrocyte culture is liquid, it is difficult to feed it directly to livestock, and must be mixed with livestock feed. The water in the feed to increase the liquid culture medium mixing the feed and it contains a nutrient liquid culture medium itself rich liquid Tino My three tests (Actinomycetes) the germs of a series or Aspergillus (Aspergillus) sequence can be easily breed common Contamination by various bacteria, which is a problem of the solid culture method, becomes a problem. Therefore, to mix the liquid culture medium with the feed, the feed must be thoroughly sterilized so that contamination by various germs is not a problem. However, the characteristics of the medium developed by the present invention is very high density of red yeast bacteria and at the same time the secondary metabolic process is maximized, it contains very rich secondary metabolites. In other words, because the density of red yeast bacteria in the liquid medium is very high, when mixed with feed, the red yeast bacteria are already distributed in sufficient amounts in the feed, which leaves other bacteria free to coexist. In addition, the secondary metabolite of hongguk bacteria because it is antibacterial to prevent the growth of other germs. Therefore, using the liquid medium developed by the present invention can be easily used as a chicken feed without the sterilization process of the feed. The following describes an example of a culture medium cultured with maximal secondary metabolite and microorganism density produced using a liquid culture medium and a liquid culture method developed by the present invention.

1) spawn culture

In the spawn culture process, a culture method generally known as spawn culture of Hongguk can be used. The purpose of this spawn propagation process is to cultivate microorganisms while supplying enough oxygen in a liquid medium rich in carbon to promote the growth and proliferation of red yeast bacteria while suppressing secondary metabolic processes. In one embodiment of the present invention, Monascus purpureus (ATCC 34892) was used for spawn propagation. Monascus purpureus ATCC 34892 used for spawn growth were first incubated in PDA agar (300g Diced potato, 20g glucose, 15g agar / ℓ) medium for 5-8 days to obtain spores using distilled water. Mixed with 20% glycerol solution to make spawn seedling spawn. When spore seed spawns are made, this spore seed spawn seed contains 40 ml of YEME (3 g yeast extract, 5 g peptone, 3 g malt extract, 340 g sucrose, 10 g glucose, 1 g MgCl 2 · 6H2O) medium per 250 ml of flask for sufficient oxygen supply. Inoculate 0.2% (200 μl spore seed spawn per 100 mL growth medium) in a baffled flask and grow for 2-4 days. If bioreactors are used, there are primary seed cultures and secondary seed cultures. In the first seed culture, put 1.5 L YEME medium in a 5 L aspirator bottle, and incubate 3.5 × 10 9 spores for 1 ~ 2 days according to the culture conditions of each microorganism. Secondary seed culture in the growth medium is supplied to the 150L bioreactor 60L growth medium, and then 10% of the primary seed culture medium is incubated at 150 rpm for 1-2 days in the range of pH 6.5-7.2.

2) Cultivate secondary metabolite production

After growing hongguk bacteria through a common seed culture method, the liquid culture method using the hongguk culture medium of the present invention enters the culture process to promote the production of secondary metabolites and increase the bacterial density.

In this culturing process, one can use one of the culture media of the present invention, specifically, the medium described in Table 3, but is not limited thereto.

In this culturing process, preferably, a baffle flask or the like is sufficiently supplied with oxygen to incubate for 6 to 8 days.

In one embodiment of the present invention, 250 ml of production medium was supplied per liter of the baffle flask, and the stirring speed was incubated at 150 rpm for 6 to 10 days. When using a bioreactor, 775 L of the production medium was supplied to 1,000 L of bioreactor, and 6% of the secondary seed culture was inoculated and incubated at 80 to 120 rpm for 6 to 10 days in the pH range of 5.8 to 6.3.

Use of S. aureus cell culture medium

Cell culture medium of hongguk bacteria obtained by the hongguk culture method of the present invention as described above can be used as an animal feed additive as an active ingredient for lowering cholesterol of livestock.

When using the cell culture liquid of hongguk bacteria as an animal feed additive, preferably the cell culture liquid is added to the animal feed and aged for a certain time to make the animal feed for lowering cholesterol of the livestock. More preferably, 1 ~ 10% of the honggukyun cell culture medium is added to the livestock feed is aged for 3 days or more to make the livestock feed.

Low-cholesterol livestock can be produced by administering a livestock feed comprising the erythrocyte bacterial culture medium of the present invention to a livestock, and more preferably, the livestock feed is administered to the livestock at least once a day and at least two days to obtain a low-cholesterol livestock. To do that.

Low cholesterol livestock herein is understood to mean livestock with significantly lower cholesterol levels compared to normal average livestock species, unless specifically limited. Low cholesterol meat products, low cholesterol eggs, low cholesterol milk, dairy products and the like are also construed as the same meaning.

Low-cholesterol meat products, low-cholesterol eggs, low-cholesterol animal foods such as low-cholesterol milk can be obtained from the low-cholesterol livestock produced in the above manner. In addition, the primary product can be used to produce a variety of low cholesterol dairy products such as yogurt, butter, cheese, powdered milk, or a variety of low cholesterol processed foods using eggs, meat, and the like.

In addition, according to the effectiveness of the well-known red yeast fungi cell culture medium of the present invention can be applied to humans. Therefore, another aspect of the present invention relates to a functional food comprising the Hong Kong bacteria culture medium as an active ingredient for lowering cholesterol, and also relates to a hyperlipidemia therapeutic agent comprising the Hong Kong bacteria culture medium as an active ingredient for lowering cholesterol.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited by the following examples, and those of ordinary skill in the art to which the present invention pertains should be within the equivalent scope of the technical concept of the present invention and the claims to be described below. Of course, various modifications and variations are possible.

Example 1

Secondary Metabolite and Hongguk Density Optimization Experiment

a) Comparison of productivity of secondary metabolites

After culturing red yeast bacteria in each secondary metabolite production medium to maximize the secondary metabolite of the present invention and the microorganism density, 5 ml of the culture solution was obtained from each culture medium and filtered with a 0.45 μm filter. The filtrate was extracted with the same amount of ethyl acetate, and then the absorbance of the red pigment was measured at 480 nm using a spectrophotometer to compare the productivity of the secondary metabolite. (1.0 absorbance at OD480nm is 15mg of pigment per liter) The results are shown in Table 4 below.

Production medium Secondary metabolite production (per liter) Remarks PST 120.1 mg Badge of the invention PRT 125.4 mg PCT 83.6 mg PPT 103.6 mg PWT 111.5 mg SP 39.8 mg Conventional liquid medium YE 35.7 mg MY 45.2 mg

b) microbial density comparison

The comparison of the density of S. aureus in the secondary metabolite production medium was compared by measuring the dry weight of S. aureus produced in different media. Hongguk culture medium produced in each production medium of step a) above was filtered with a 0.8 μm nylon filter and dried in vacuo at 60 ° C. and then weighed. The results are shown in Table 5 below.

Production medium Dry Cell Weight (per liter) Remarks PST 30.5 g Badge of the invention PRT 31.2 g PCT 21.3 g PPT 25.1 g PWT 27.8 g SP 18.6 g Conventional liquid medium YE 13.8 g MY 14.7 g

As can be seen from the above results (Table 4, Table 5), the medium developed in the present invention was more than three times the secondary metabolite production and two times higher than the hongguk density compared to other media that are currently utilized. Especially, PST production medium and PRT production medium showed higher productivity of secondary metabolite and density of red yeast bacteria than other production medium. Therefore, Example 2 and Example 3 were performed using this PST production medium and PRT production medium.

Example 2

Low-Cholesterol Eggs Produced by Using a Hong Kong Bacteria Culture Cultured with PST Production Medium and PRT Production Medium as Feed Additives

① Mixing and feeding of Hongkuk culture medium and laying hen feed

Hongguk culture medium produced through the above-described process was mixed with commercially available laying hens at 1%, 5%, 15% and 30% and fed to chickens every 12 hours. The chickens used in the experiment were 33 weeks old, and 6 animals per cage were divided into a no addition group, a 1% addition group, and a 5% addition group for 21 days.

② Cholesterol content analysis of eggs

The cholesterol content analysis of eggs was carried out by the following method.

First, boil the eggs completely, separate the egg yolk, add 15 ml of chloroform: methanol (2: 1, v / v) solution per 1 g of egg yolk, homogenize it with a grinder, and grind for 30 seconds using ultrasonic waves. It was filtered by a filter. 0.1 ml of this filtrate was saponified with 0.3 ml of 33% KOH (in H 2 O) and 3 ml of 95% Ethanol at 60 ° C. for 15 minutes. After the saponification reaction, cholesterol was extracted with 10 ml of Hexane and 1 ml of the Hexane layer was evaporated in another test tube. After the reaction with cholesterol esterase, cholesterol oxidase, and peroxidase in the test tube, the absorbance at 500 nm was measured with a spectrophotometer, and the cholesterol content in the egg was calculated by comparing with the standard curve.

Cholesterol content of the eggs obtained after the 21-day breeding by adding the Hong Kong cell culture medium of the present invention cultured using a PST production medium to chicken feed is shown in Table 6 and FIG.

Feed rate Cholesterol Content / Egg Yolk (mg / g) % Degradation 0% 16.74 0 One% 13.16 21.4 5% 11.87 29.1 15% 11.76 29.8 30% 11.62 30.6

The results of administration of the culture medium cultured using the PRT production medium instead of the PST production medium for 21 days are shown in Table 7 and FIG. 2.

Feed rate Cholesterol Content / Egg Yolk (mg / g) Degradation rate (%) 0% 16.74 0 One% 13.02 22.3 5% 11.78 29.7

As a result of measuring the change in cholesterol content of the eggs obtained after feeding the feed containing the erythrocyte culture medium cultured in PST production medium and PRT production medium (Table 6, Table 7, Fig. 1, Fig. 2), In the case of the addition of the% erythrocyte culture medium, the cholesterol content of the eggs was decreased by 21.4% on average, and the cholesterol content of 29.1% was decreased in the 5% hongkong bacteria culture group. In addition, 29.8% in the 15% S. aureus medium and 30.6% in the 30% S. aureus group added cholesterol, but this was not significantly different from that of the 5% group. It can be interpreted as indicating a range. In the case of the PRT production medium, the cholesterol content in the eggs was decreased by 22.3% in the group added with 1% S. aureus and 29.7% in the group containing 5% S. aureus, which was almost similar to or slightly higher than that of PST medium.

Example 3

Low-cholesterol broiler produced by using erythrocyte culture medium cultured using PST production medium and PRT production medium as feed additive

① Mixing and feeding of Hongkuk culture medium and broiler feed

The hongguk culture medium produced in Example 1 was mixed with a commercial broiler feed and fed to chickens every 12 hours. The chickens used in the experiment were 6 weeks old, and 6 animals per cage were divided into no addition group, 1% addition group, and 5% addition group for 20 days, and blood was collected from each broiler to measure blood cholesterol levels.

② Analysis of blood cholesterol content in broilers

The blood cholesterol content analysis of broilers was performed by the following method.

Serum was separated from broiler blood and 0.1 ml of serum was saponified with 0.3 ml of 33% KOH (in H 2 O) and 3 ml of 95% Ethanol at 60 ° C. for 15 minutes. After the saponification reaction, cholesterol was extracted with 10 ml of Hexane and 1 ml of the Hexane layer was evaporated in another test tube. After the reaction was performed using cholesterol esterase, cholesterol oxidase, and peroxidase in the test tube, the absorbance was measured at 500 nm using a spectrophotometer, and blood cholesterol content was calculated by comparing with a standard curve.

Table 8 shows the blood cholesterol content of broiler broilers produced by breeding erythrocyte culture medium cultured using PST production medium and fed for 20 days.

Feed rate Blood cholesterol content (mg / dl) Degradation rate (%) 0% 144.8 0 One% 121.4 16.2 5% 101.5 30

In addition, the results of administering the erythrocyte culture medium cultured using PRT medium instead of PST medium are shown in Table 9.

Feed rate Blood cholesterol content (mg / g) Degradation rate (%) 0% 144.8 0 One% 120.8 16.6 5% 103.7 28.4

The results of measuring the blood cholesterol level of broiler chickens were added to broiler feed by adding the erythrocyte culture medium cultured in PST production medium and PRT production medium (Table 8 and Table 9). Showed a decrease in the blood cholesterol level, and 30% of the cholesterol content of the 5% hongguk culture medium was decreased. In the case of PRT production medium, the cholesterol content of 16.6% was added in the group added with 1% S. aureus and 28.4% in the group containing 5% S. aureus.

Example 4

Low-Cholesterol Milk Produced by Using a Hong Kong Bacteria Culture Cultured with PST Production Medium and PRT Production Medium as Feed Additives

The erythrocyte culture medium produced by the same process as in low cholesterol eggs was mixed with commercially available dairy feed at 1% and 5% and fed to cows every 12 hours. The cows used in the experiment were 25 months of age, and each group was fed four week-old cows into the no-added, 1% and 5% groups, and fed twice a day for three weeks. Milk cholesterol levels were measured. In the measurement method, 0.1 ml of milk was saponified with 0.3 ml of 33% KOH (in H 2 O) and 3 ml of 95% Ethanol at 60 ° C. for 15 minutes. After the saponification reaction, cholesterol was extracted with 10 ml of Hexane and 1 ml of the Hexane layer was evaporated in another test tube. After the reaction was performed using cholesterol esterase, cholesterol oxidase, and peroxidase in the test tube, absorbance at 500 nm was measured using a spectrophotometer, and the cholesterol content in milk was calculated by comparing with a standard curve.

Table 10 shows the results of administering the erythrocyte culture medium cultured using the PST production medium.

Feed rate Blood cholesterol content (mg / dl) Degradation rate (%) 0% 10.16 0 One% 8.09 20.4 5% 7.87 22.6

Table 11 shows the results of administering the erythrocyte culture medium cultured using the PRT production medium instead of the PST medium.

Feed rate Blood cholesterol content (mg / g) Degradation rate (%) 0% 10.16 0 One% 7.98 21.5 5% 7.56 25.6

As a result of measuring the cholesterol level of milk by adding red pepper bacteria cultured in PST producing medium and PRT producing medium to cow's feed (Table 10 and Table 11), average 10.4% of red pepper culture medium added in PST producing medium The milk cholesterol content was decreased, and the cholesterol content of 22.6% was decreased in the 5% hongkuk culture medium added group. In the case of the following PRT production medium, milk cholesterol content was decreased by 21.5% in the group added with 1% S. aureus and 25.6% in the group containing 5% S. aureus.

As can be seen from the above examples, according to the present invention, while maximizing the production of secondary metabolites of the red yeast bacterium having a cholesterol lowering effect, at the same time, the liquid medium which can maximize the red yeast bacterium density in the culture medium has a short production cost. It is possible to obtain a cell culture medium of hongguk bacteria that can be used as a livestock feed additive within a time, and to obtain various low-cholesterol animal foods such as low cholesterol, low cholesterol broiler, low cholesterol milk, etc. In addition, the cell culture medium of the hongguk bacterium of the present invention can be used as a functional food and hyperlipidemia treatment for humans according to the known hongguk bacteria.

Claims (17)

In liquid culture medium of Hongkukyun consisting of nitrogen source, carbon source, and trace elements essential for bacterial growth, For the Secondary Metabolic Process Activation and Increasing Bacterial Density (a) proflo, a cottonseed extract, as a nitrogen source; (b) a material selected from the group consisting of sugar, rice flour, corn flour, potato flour, flour and mixtures thereof as a carbon source; (c) sodium (Na), calcium (Ca), iron (Fe), copper (Cu) and manganese (Mn) components as trace elements; Hongkuk culture medium comprising a. According to claim 1, wherein the hongguk culture medium Propflow 0.5-1.5%, Carbon source 1.5-4%, NaCl 0.1-0.5%, CaCO₃0.1-0.5%, FeCl ₃ · 6H ₂ O 0.01-0.04%, CuCl ₂ · 2H ₂ O 0.001-0.002%, MnCl ₂ 4H ₂ O 0.001 to 0.002%, ZnCl ₂ 0.002 to 0.006%, Na ₂ B ₄ O ₇ 10H ₂ O 0.001 to 0.002%, (NH ₄ ) ₆ Mo ₇ O ₂₄ 4H ₂ O 0.001 to 0.002% and distilled water Hongkuk culture medium comprising the remainder. A culture method of Hongguk characterized in that the culture of the spawn cultured Honggukyun in the culture medium of claim 1 to obtain a cell culture solution containing a large amount of secondary metabolites. The method of claim 3, Seed cultured inoculated into the culture medium of the seed culture medium of claim 1, characterized in that for 6 to 8 days incubation under aerobic conditions. Cell culture medium of hongguk bacteria obtained by the culture method of claim 3 or 4. Livestock feed additive comprising a hongguk cell culture medium of claim 5 as an active ingredient for lowering cholesterol of livestock. The method of producing a livestock feed, characterized in that the fermentation of hongguk cell culture medium of claim 5 added to the livestock feed. 8. The method of claim 7, characterized in that 1 to 10% of the hongkuk cell culture medium is added to the livestock feed and aged for at least 3 days. A method for producing low-cholesterol livestock, comprising administering to a livestock feed comprising livestock culture medium according to claim 5 cell culture. The method of claim 9, The livestock feed is administered to the livestock at least once a day, at least two days. 10. The method of claim 9, wherein said low cholesterol livestock has a cholesterol level of at least 10% lower than a normal mean livestock species. A low cholesterol meat product obtained from a livestock produced by the method of claim 9. Low cholesterol eggs obtained from livestock produced by the method of claim 9. Low cholesterol milk obtained from livestock produced by the method of claim 9. Dairy products using low cholesterol milk of claim 14. Functional food containing the hongguk cell culture medium of claim 5 as an active ingredient for lowering cholesterol. Hyperlipidemia treatment agent comprising the honggukyun cell culture medium of claim 5 as an active ingredient for lowering cholesterol.