KR20140006509A - Bifidobacterium longum strain producing conjugated linoleic acid and uses thereof - Google Patents

Abstract

The present invention provides a Bifidobacterium longum CGB-C11 (deposition number KCTC 11979BP) strain, which is a strain producing conjugated linoleic acid (CLA). The strain of the present invention has excellent CLA production capacity, and strong resistance to acid including gastric acid and bile and an antibiotic. A composition containing the strain is formed into a capsule containing the strain and CLA and coated with a coating material formed of a dairy product, powder, and water soluble polysaccharide, and can be applied to food, drink, animal feed, functional food, and medicines.

Description

Bifidobacterium longum strain producing conjugated linoleic acid and its use {Bifidobacterium longum strain producing conjugated linoleic acid and uses thereof}

The present invention relates to a method for producing conjugated linoleic acid (CLA), which comprises producing Bifidobacterium The present invention relates to a method for producing conjugated linoleic acid, which comprises producing a conjugate linoleic acid comprising a microorganism preparation, a capsule, a functional food, a composition for animal feed and a pharmaceutical composition, and a step for culturing the strain, ≪ / RTI >

Conjugated linoleic acid (CLA) is a co-liquefied isomer of linoleic acid (LA), an essential fatty acid, and a natural fatty acid component found in trace amounts in milk or muscle of ruminants. CLA has a conjugated double bond in the cis-9, trans-11 or trans-10 and cis-12 positions in the chain, and particularly has covalent double bonds in the cis-9 and trans- Isomer exhibits a useful physiological activity in the human body.

CLA has been shown to reduce the incidence of arteriosclerosis (Artery, 1997. 22: 266-277), enhance immune function (J. Nut. 1999. 129: 32-38), anticancer activity (Anticancer research 1997. 17: 969-973) , J. Am. J. Physiol., 1998. 275: R667-R672), which shows excellent therapeutic effects against diseases such as diabetes, obesity, It is also known to inhibit. Due to these properties, CLA can be usefully used as an active ingredient of functional foods and medicines.

CLA is mainly contained in animal foods, and is known to exist in a large number of ruminants. The amount of CLA in beef is 2.9 ~ 4.3 ㎎ CLA / fat, 5.6 ㎎ CLA / fat in lamb and 0.3 ~ 0.6 ㎎ CLA / fat in seafood. The daily intake of CLA in humans is expected to be 0.1 g / day for vegetarian Oriental and 0.4 g / day for Western-style carnivores.

To date, several methods have been developed for mass production of CLA. Conventional methods include chemically synthesizing CLA from LA such as urea addition method, molecular distillation method, and HPLC method, and a number of microorganisms having activity to convert LA to CLA have been isolated. However, chemical synthesis methods require expensive equipment or take too much time to process. In addition, since these methods produce not only a single kind of CLA but also various kinds of isomers together, it is very inefficient to produce a single kind of CLA isomer by conventional chemical methods.

Therefore, the most efficient way to produce CLA is to produce and isolate CLA from microorganisms capable of producing CLA. Representative microorganisms known to produce CLA include Lactobacillus , Propionibacterium , Butyrivibrio < RTI ID = 0.0 > fibrisolvens ). In many countries, they are usefully used as active ingredients of functional foods such as probiotics, feeds, and medicines administered to livestock.

Unlike the chemical synthesis method in which the ratio of cis-9, trans-11, trans-10, and cis-12 is about 50%, CLA synthesized by intestinal microorganisms is cis- -11 < / RTI > In fermented dairy products using skim milk, very low cis-9 and trans-11 were detected only in small quantities of cis-9 and trans-11 in ruminant-derived fats, , It is known that trans-11 does not exist. Thus, Korean Patent Application No. 2001-0047292 discloses a method of adding pure cis-9 or trans-11 type CLA synthesized by a microorganism to a food using a microorganism of the genus Lactobacillus, Only the content has been confirmed.

In addition, in order for the CLA-producing microorganism to be used as an active ingredient of foods or medicines, the microorganisms must remain at a high rate similar to that during the production of the product throughout the distribution period of the manufactured product, Intestine) should have high survival rate and activity. In addition, excellent resistance to antibiotics is required to compete with harmful bacteria in the intestines, including superbacteria.

However, many of the microorganisms used as active ingredients in the currently marketed products were not able to remain in storage during the storage as well as in the product during storage, and the resistance to antibiotics was weak. Therefore, in addition to excellent CLA production ability, an invention for a strain which can exhibit high viability and activity in the body of an animal and can directly add the cell itself as an active ingredient of foods and medicines remains a challenge.

Korean Patent Registration No. 0715730 discloses a novel Bifidobacterium longum A24 having an antimicrobial activity against harmful pathogenic microorganisms, but is different from the Bifidobacterium longum strain producing the conjugated linoleic acid of the present invention.

The present invention relates to a strain of Bifidobacterium longum CGB-C11 (Accession No. KCTC 11979BP), which is excellent in the ability to produce conjugated linoleic acid isolated from feces and has excellent tolerance to acid and antibiotics, It is an object of the present invention to provide a probiotic microbial agent prepared in the form of a dairy product, a food, a drink, an animal feed, a functional food and a medicine and a composition thereof, a powder or a capsule form using the strain.

In order to solve the above-mentioned problems, the present invention relates to a method for producing conjugated linoleic acid ( Bifidobacterium longum strain.

The present invention also provides a microorganism preparation for producing a conjugated linoleic acid comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a capsule containing the strain as an active ingredient and containing a water-soluble polysaccharide as a covering substance.

The present invention also provides a functional food comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a composition for animal feed comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a pharmaceutical composition for preventing or treating cancer, arteriosclerosis, diabetes or obesity comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a method for producing a conjugated linoleic acid comprising culturing the strain.

The strain of the present invention not only can produce CLA from LA with excellent efficiency, but also has excellent resistance to gastric acid, bile acid, and antibiotics. Since the strain of the present invention can accumulate CLA in the cells after production, there is an effect that CLA is indirectly produced. In addition, the strain of the present invention can be effectively used for biosynthesis of large amounts of CLA isomers in addition to CLA by biological methods such as cell immobilization.

1 shows the results of GC analysis for confirming the composition of a fatty acid produced in a medium to which linoleic acid (LA) is added in the strain of the present invention.
Fig. 2 is a graph showing the growth state, the degree of CLA production, and the pH change of the culture solution provided by the strain of the present invention in a medium supplemented with linoleic acid (LA).
3 is a graph showing the growth state and the degree of CLA production provided by the strain of the present invention in a medium to which no linoleic acid (LA) is added.
Fig. 4 is a graph showing the amount of CLA distributed in the culture medium and the cells when the strain of the present invention was cultured in the medium supplemented with linoleic acid (LA).
FIG. 5 is a graph comparing the amount of CLA distributed in the culture medium and the cells when the strain of the present invention was cultured in a medium to which no linoleic acid (LA) was added.
FIG. 6 shows the results of culturing the strain according to the present invention in a medium supplemented with glucose, fructose, lactose and sucrose, respectively, to determine the degree of growth (a) and the amount of CLA produced (b) Fig.

In order to accomplish the object of the present invention, the present invention provides a method for producing conjugated linoleic acid (CLA), which comprises producing Bifidobacterium longum strain.

The strain was isolated and selected by inoculating human feces into MRS (Man Rogosa Sharpe) medium, which is a Bifidobacterium selective agar medium, and culturing under anaerobic conditions. The strain is characterized in that it has at least one of acid resistance and biliary cholestasis and is resistant to antibiotics. The antibiotic is preferably tetracycline, streptomycin or kanamycin, but is not limited thereto. Preferably, the strain is Bifidobacterium ronggeom (Bifidobacterium longum ) CGB-C11 (Accession No. KCTC 11979BP).

CLA produced from the above strains contains only the three-dimensional structure of cis-9 and trans-11. Since these are isomers having various physiological activities as known in the art, fatty acids including these structures and acylglycerols can be obtained from various animal- Can be widely used for the development of dairy products, dairy products, fermented foods, health functional foods, and probiotics by raw materials of dairy products and plants, as well as for efficient biosynthesis of large amounts of isomers of CLA by biological methods Can be used.

The present invention also provides a microorganism preparation for producing conjugated linoleic acid comprising the strain of the present invention or a culture thereof as an active ingredient. The strain is preferably Bifidobacterium longum, and more preferably Bifidobacterium longum CGB-C11.

The present invention also provides a functional food comprising the strain of the present invention or a culture thereof as an active ingredient. The strain is preferably Bifidobacterium longum, and more preferably Bifidobacterium longum CGB-C11.

In addition to the active ingredient, the functional food of the present invention may further contain various auxiliary ingredients as necessary. In the case of the functional food of the present invention, vitamins such as vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B12, folic acid, vitamin C, vitamin D3 and vitamin E, Iron, magnesium, potassium, zinc, etc., or lactic acid bacteria.

In addition, among the functional foods of the present invention, health drinks may contain various flavors or natural carbohydrates as additional components such as ordinary beverages. Examples of the flavoring agent include natural sweetening agents such as tau Martin and stevia extract, and synthetic sweetening agents such as saccharin and aspartame. Examples of natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol.

The present invention also provides a composition for animal feed comprising the strain of the present invention or a culture thereof as an active ingredient. The strain is preferably Bifidobacterium longum, and more preferably Bifidobacterium longum CGB-C11.

The feed composition of the present invention is added to the base feed at a certain ratio. The basic diet may be composed of corn, soybean meal, whey, fish meal, molasses, salt, vitamin premix, and mineral premix. The vitamin premix can be composed of vitamin A, vitamin D, vitamin E, riboflavin and niacin, and the mineral premix can be composed of manganese, iron zinc, calcium, copper, cobalt and selenium.

The present invention also provides a pharmaceutical composition for preventing or treating cancer, arteriosclerosis, diabetes, or obesity comprising the strain or the culture liquid of the present invention as an active ingredient. Since the pharmaceutical composition contains conjugated linoleic acid, it is possible to prevent or treat diseases controlled by conjugated linoleic acid. The strain of the present invention produces CLA, and the produced CLA exhibits a decrease in the incidence of arteriosclerosis (Artery, 1997. 22: 266-277), immune function improvement (J. Nut. 1999. 129: 32-38) (Anticancer research 1997. 17: 969-973), and it is also known that it exhibits excellent therapeutic effect against diseases such as diabetes and suppresses obesity through reduction of body fat (AM, J. Physiol., 1998. 275: R667-R672) Therefore, the strain of the present invention or a culture thereof can be used as a pharmaceutical composition for preventing or treating cancer, atherosclerosis, diabetes or obesity. The strain is preferably Bifidobacterium longum, and more preferably Bifidobacterium longum CGB-C11.

The dosage or amount of the pharmaceutical composition of the present invention is 60-130 uM (Cancer Epidemiol. Biol. Prev. 2000. 9: 689-696), and the dosage or amount of functional food is 3.4-6 g / day (J. Nutr. 2000. 130: 2943-2948), but it can be added or subtracted as needed. The pharmaceutical composition of the present invention can be safely absorbed into the body regardless of the intake amount. The dosage or amount may vary depending on factors such as the type and amount of the active ingredient and the other ingredients contained in the pharmaceutical composition, the type of the formulation and the age, body weight, general health condition, sex and diet, time of administration, Duration of administration, drug used concurrently, and the like. The pharmaceutical composition of the present invention is free from side effects when administered to human body as compared to chemically synthesized CLA-containing foods and medicines.

The food or pharmaceutical composition of the present invention may be formulated to contain one or more carriers acceptable for pharmaceutical or food addition to the above-mentioned effective ingredients. The pharmaceutical or food acceptable carrier may be a mixture of saline, sterilized water, Ringer's solution, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components. If necessary, an antioxidant, Other conventional additives such as a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease, or according to each ingredient, using the method disclosed in the appropriate field in the art, or the method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA.

Formulations of the pharmaceutical compositions of the present invention may be granules, powders, granules, coated tablets, tablets, capsules, sticks, extracts, suppositories, syrups, juices, suspensions, emulsions, have.

The capsules of the present invention are composed of a coating material and a core material to be coated. The nucleus material in the capsules of the present invention preferably contains the strain of the present invention together with linoleic acid. In the capsules of the present invention, the coating material surrounding the nuclear material uses a water-soluble polysaccharide having excellent water absorption, dispersibility, and stickiness.

The water-soluble polysaccharides usable in the present invention include starch, agar, carrageenan, alginic acid, sodium alginate, polymethylmethacrylate, wheat protein, soy protein, methylcellulose, hydroxypropylcellulose cellulose derivatives such as hydroxypropylcellulose and hydroxypropylmethylcellulose, xanthan gum, arabic gum, locust bean gum, guar gum, tamarind A gum such as tamarind gum, Tara gum, karaya gum, tragacanth gum, ghatti gum, and gellan, xanthan, pectin, (LM, HM type), dextran, glucan, glucomannan, arabino galactan, furcelleran, pullulan, glucosamine, Gelatin, casein (c asein).

The amount of the coating material in the preparation of the capsules of the present invention may be appropriately adjusted depending on the application and purpose, and it is preferable to use the coating material in an amount ranging from 1 to 80% by weight, which is generally used on the basis of nuclei Do. The capsules of the present invention may further contain an emulsifying agent, a protecting agent and a plasticizer as necessary. The capsule preparation of the present invention may be prepared by using an appropriate one of the commonly used encapsulation methods. The composition containing the capsules of the present invention can be specifically used as dairy products (milk, soy milk, processed milk), fermented milk (liquid yogurt, yogurt yogurt), fermented foods (kimchi, .

The present invention also provides a method of producing a conjugated linoleic acid comprising culturing a strain of the present invention. The strain is preferably Bifidobacterium longum, and more preferably Bifidobacterium longum CGB-C11. As a method for culturing the strain of the present invention, a method commonly used in the art can be used. Suitable carbon sources in the strain culture of the present invention can be used, such as glucose, fructose, lactose and sugar, but lactose is preferable because it produces the maximum CLA.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  1: Strain isolation and identification of the present invention

To isolate CLA-producing microorganisms, 10 fecal samples from Korean adults were collected and inoculated into MRS (Man Rogosa Sharpe) medium supplemented with 0.05% L-cystein. The cells were diluted with sterile physiological saline 0.85% (w / v) to give a colony of about 30 to 150, and stained with Bifidobacterium selective agar. Then, a gas pack (MGC, mitsubishi) and cultured at 37 ° C for 72 hours. At this time, linoleic acid (LA) was well-emulsified in 0.5% (w / v) tween 80 (tween 80) and added to MRS medium to which 0.05% of L-cystein was added.

Approximately 300 bacterial colonies were randomly selected from the cultured medium, and each strain was subcultured twice in MRS medium, then inoculated in a 20 ml test tube and incubated for 48 hours. The culture was extracted with hexane and the absorbance at 233 nm was measured to obtain the amount of CLA, and the amount of CLA produced was corrected by adjusting the amount of absorbance at 600 nm.

The number of viable cells was measured by counting the number of bacterial colonies formed after 72 hours with a gas pack in an anaerobic tank (Difco, USA) by using 10 times dilution method using BS agar medium. Thus, a strain having excellent CLA production ability was selected and named as CBG-C11. The nucleotide sequences of rRNA were analyzed for the strains of the present invention and the genus and species thereof were identified. As a result, the CBG-C11 strain was found to belong to Bifidobacterium longomi. This strain was deposited with CBG-C11 (KCTC 11979BP) on July 6, 2011 at the Genetic Resource Center, Korea Research Institute of Bioscience and Biotechnology.

Example  2: Confirmation of the composition of the fatty acid produced by the strain of the present invention

In order to confirm the composition of the fatty acid produced by the strain of the present invention and to confirm again whether the strain of the present invention produces CLA, the following GC was performed. The cells were cultured in a medium containing LA (500 / / ml) for 42 hours and centrifuged. To the cells suspended in the culture broth or distilled water, isobutanol (2-fold volume) was added and vigorously mixed. 1.5-fold volume of hexane was added thereto and mixed with shaking for 3 minutes. The mixture was centrifuged at 3000 rpm for 5 minutes at room temperature, and the absorbance of the supernatant was measured at 233 nm. The extracted fatty acids were [American Oil Chemists Society: Official Method and Recommended Practice pf AOCS, 4th. ed. (1989)] to prepare a sample required for GC analysis.

GC analysis was performed using GC DS-6200 (DONAM) with FID attached. The column was an HP-FFAP capillary column (30 m × 0.25 mm, thickness 0.25 μm) injector temperature was 250 ° C, and the detector temperature was 270 ° C. The carrier gas was helium, eluting at a flow rate of 1 ml / min, and a split ratio of 50: 1. The area of each peak was determined using an integrator (moder 3390A, Hewlett-Packard, USA) connected to the instrument. The identification of CLA was confirmed by comparing with the retention time of the reference material, and the content of CLA was used as the analytical sample by the area of the reference material and the area ratio of CLA.

As a result, as shown in Fig. 1, LA and CLA peaks were observed. Therefore, it was found that the strain of the present invention can produce CLA using LA as a substrate.

Example  3: When the strain of the present invention was cultured CLA  Identify optimal conditions for generation

In order to confirm the optimum conditions for CLA production of the strain of the present invention, growth characteristics were observed under two conditions of the presence or absence of LA as a substrate. First, the inoculated strain was inoculated to 2 liters of MRS medium containing LA (500 μg / ml) containing 1% of the activated culture, and the culture was recovered by time to measure the cell concentration, the amount of CLA and the pH of the culture The results are shown in Fig. As shown in FIG. 2, the amount of CLA produced increased with the shift to the log phase, and the maximum CLA was generated just before reaching the stationary phase. The initial pH of the culture was about 6.5, but after fermentation for about 42 hours, the pH was about 4.2.

On the other hand, the pre-cultured strain was inoculated to the MRS medium containing no LA in an amount of 1%, and the growth curve was measured by dividing into 20 ml test tubes and measuring the pH of the culture every 3 hours. The cells were recovered and suspended in a Tris buffer solution having a volume of 10 times the amount of the cells. The enzyme reaction was performed to observe the change in the activity of CLA isomerase according to the change of the growth curve. At this time, the concentration of LA added to the enzyme reaction was 100 占 퐂 / ml, and the results are shown in Fig. As shown in FIG. 3, when LA was added without culture, the time required to reach the stationary phase was longer than that when LA was added and cultured. The final pH of the culture was about 4.2. Therefore, it was found that the strains of the present invention produce CLA at a maximum immediately before transition from the spring to the stand.

In addition, the results obtained in the presence of LA indicate that when the product is developed by fermenting the bacteria in the presence of the substrate and when the LA is not present, the result obtained is that the CLA is indirectly added , It can be used to determine the growth period of the cells showing the maximum titer per unit cell.

Example  4: When the strain of the present invention is cultured, CLA The amount of

In order to confirm the amount of CLA distributed in the culture medium and the cells in the culture of the strain of the present invention, the following experiment was conducted. First, CLA production was measured 18, 24, 36, and 42 hours after culturing for 48 hours after inoculation with the strain containing LA. The results are shown in FIG. As shown in FIG. 4, the ratio of CLA present in the cells and the culture solution was found to be 1: 1.5 after 42 hours as shown in Table 1, and the total CLA conversion rate was 45.6%.

The CLA conversion rate of the strain of the present invention Strain name division Production rate
CLA (ug) / dry cell amount (mg) CLA Conversion Rate (%) Distribution ratio (%) CBG-C11
Culture solution 113.9 45.6
59.4 Mycelium 73.8 38.5

On the other hand, the strains were inoculated into the culture solution without LA, collected for 18, 24, 36, and 42 hours, mixed with the buffer solution containing LA, and reacted for 1 hour. The distribution was measured and is shown in FIG. As shown in Fig. 5, the amount of CLA increased with the lapse of growth time. After 36 hours of culture, as shown in Table 2, the distribution of CLA produced by the cells for 1 hour was about 1: 1 in the ratio of the cells to the culture solution.

The CLA conversion rate of the strain of the present invention Strain name division Production rate
CLA (ug) / dry cell amount (mg) CLA Conversion Rate (%) Distribution ratio (%) CBG-C11
Culture solution 24.5 51.2
51.0 Mycelium 23.5 49.0

Therefore, the strain of the present invention can secrete CLA into a medium and accumulate it in the cells. This property of the strains of the present invention can be used to produce CLA by adding the cells themselves as an active ingredient in foods and medicines.

Example  4: Strain culture of the present invention Carbonic  Growth and growth of microorganisms CLA  Change in yield

In the culture of the strain of the present invention, in order to confirm the growth and the change of CLA production depending on the species of the glycogen, various mediums such as glucose, fructose, lactose and sucrose were added, The strain was cultured to measure the degree of growth and the amount of CLA produced. The results are shown in FIG.

As shown in FIG. 6 (a), the most suitable carbon source for growth of each strain was glucose and lactose when LA was present in the medium, and lactose and glucose when LA was not present in the medium. On the other hand, the amount of CLA produced after culturing in LA supplemented medium for 48 hours showed a maximum value when lactose was used as a carbon source, as shown in FIG. 6 (b).

Example  5: of the strain of the present invention pH , Resistance to bile and antibiotics

The characteristics of the strain of the present invention such as pH tolerance, bile resistance and antibiotic resistance were confirmed as follows. This study was carried out to investigate the growth characteristics of strains for use as an additive using CLA in a product fermented or inoculated with CLA - producing lactic acid bacteria using the identified new strain.

1) pH tolerance

In order to confirm the resistance of the strain of the present invention to pH, the pH of the culture solution was observed while the bacterium was cultured under the two conditions of the presence or absence of LA in the culture medium. As a result, The pH of the culture solution was drastically decreased as the bacterial growth was stopped. The final pH was reduced to about 4.2, indicating typical lactic acid fermentation pattern.

Based on the above results, the MRS medium was prepared from pH 2 to pH 6 using hydrochloric acid in order to confirm the growth of bacteria according to pH change of the culture solution. As a control, B. longum KCTC 3420 was used. The strain was inoculated in the medium and cultured at 37 ° C for 38 hours. The growth of the bacteria was observed and the results are shown in Table 3.

Strain name pH 2 pH 3 pH 4 pH 5 pH 6 CBG-C11 - +++ ++++ ++++ ++++ B. longum
(KCTC 3420) - ++ +++ ++++ ++++

(-: not proliferating at all, +: slightly proliferating, ++: usually proliferating, +++: proliferating, ++++: proliferating very much)

According to Table 3, the strain of the present invention was able to grow in a pH range of 4.0 to 6.0 and exhibited excellent pH resistance. Therefore, the strain of the present invention can grow in a wide range ranging from weak alkali to slightly acidic, and it can be fully survived on the condition that acidic environment is formed due to secretion of gastric acid.

2) bile resistance

In order to confirm the bile resistance of the strain of the present invention, MRS medium supplemented with sodium deoxycholate (0, 100, 300, 500, 800, 1000 / / ml) was prepared. Bifidobacterium longum ( Bifidobacterium longum KCTC 3420), which was distributed from the Center for Genetic Resource Research, Korea Research Institute of Bioscience and Biotechnology, was used as a control strain. The strain was inoculated into each medium and cultured at 37 ° C for 48 hours. And the results are shown in Table 4. < tb >< TABLE >

Treatment concentration (ug / ml) CBG-C11 Bifidobacterium longum KCTC 3420 0 +++ +++ 100 +++ +++ 300 +++ +++ 500 +++ +++ 800 +++ ++ 1000 ++ +

(+: Slight proliferation, ++: normal proliferation, +++: proliferation)

As shown in Table 4, the strain of the present invention was found to grow well up to 1000 / / ml of bile. Therefore, it can be seen that the strain of the present invention can stably grow in the gastrointestinal tract in which bile is secreted.

3) Antibiotic resistance

In order to confirm antibiotic resistance of the strain of the present invention, a strain cultivated overnight in a 20 ml test tube was diluted in MRS medium to prepare a solid medium. As a control strain, Bifidobacterium longum KCTC 3420 was used.

The antibiotics used in this experiment were ampicillin, tetracycline, streptomycin, rifamycin sv and kanamycin at concentrations of 50, 100, 1000, 10000 ㎍ / ㎖ Were prepared.

Each of the antibiotics prepared as described above was dropped on a paper disc by 40 μl each, and the disc was dried. The disks were placed on a germ-mixed medium and cultured at 37 ° C for 48 hours. Growth was observed and the results are shown in Table 5.

Antibiotic Class Antibiotic concentration (ug / ml) CBG-C11 Biffy Dickey Terryun Longkom KCTC 3420
Ampicillin 50 - - 100 - - 1000 - - 10000 - -
Tetracycline 50 ++ + 100 + - 1000 - - 10000 - -
Streptomycin 50 ++ + 100 + - 1000 - - 10000 - -
Rifamycin sv 50 - - 100 - - 1000 - - 10000 - -
Kanamycin 50 ++ + 100 ++ + 1000 + - 10000 - -

As shown in Table 5, the strain of the present invention showed resistance to three antibiotics. Especially, it showed tolerance to antibiotics treated with high concentrations of kanamycin and showed excellent resistance to antibiotics. Thus, it can be seen that the strain of the present invention is resistant to a wide range of species and concentrations of antibiotics.

Korea Biotechnology Research Institute KCTC11979 20110706

Claims (13)

Conjugated linoleic acid (Conjugated Linoleic acid) production ronggeom Bifidobacterium (Bifidobacterium that longum ) strains. According to claim 1, Bifidobacterium long gum ( Bifidobacterium) longum ) Bifidobacterium longgum strain, characterized in that CGB-C11 (Accession No. KCTC 11979BP). 2. The Bifidobacterium longgum strain according to claim 1, wherein the conjugated linoleic acid is cis-9, trans-11 isomer. The Bifidobacterium longgum strain according to claim 1 or 2, wherein the Bifidobacterium longgum strain has at least one property of acid resistance and bile resistance. The Bifidobacterium longgum strain according to claim 1 or 2, which is antibiotic resistant. Microbial preparation for conjugated linoleic acid production comprising the Bifidobacterium long gum strain of any one of claims 1 to 5 or a culture thereof as an active ingredient. A capsule comprising the Bifidobacterium longgum strain of any one of claims 1 to 5 as an active ingredient and a water-soluble polysaccharide as a coating material. A functional food comprising the Bifidobacterium longgum strain of any one of claims 1 to 5 or a culture thereof as an active ingredient. The functional food according to claim 8, which is selected from the group consisting of liquid yogurt, staple yogurt, cheese, kimchi and soy sauce. An animal feed composition comprising the Bifidobacterium longgum strain of claim 1 or a culture thereof as an active ingredient. A tonic tonic comprising any one of claims 1 to 5 Bifidobacterium longgum strain or its culture solution as an active ingredient. A pharmaceutical composition for preventing or treating cancer, arteriosclerosis, diabetes or obesity, comprising the Bifidobacterium longgum strain of claim 1 or a culture thereof as an active ingredient. A method for producing conjugated linoleic acid comprising culturing the Bifidobacterium longgum strain of any one of claims 1 to 5.

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