KR101953844B1 - Synbiotic composition comprising yacon extract and probiotic lactic acid bacteria - Google Patents

Abstract

The present invention relates to a food composition for assisting active intestinal proliferation of Bifidobacterium sp. Microorganisms, and to a synbiotic composition comprising said bifidobacterium microorganism and a yacon extract as a nutrient source of said strain.
More particularly, the present invention relates to a synbiotic food composition and feed additive comprising Bifidobacterium sp. Microorganism as a probiotic and a yacon extract as a prebiotic.

Description

The present invention relates to a synbiotic composition comprising yacon extract and probiotic lactic acid bacteria,

The present invention relates to a synbiotic composition comprising a prebiotic yacon extract and a probiotic lactic acid bacterium.

Intestinal microorganisms are multiplying or excreted using biocomponents separated from food and digestive tracts while maintaining mutual symbiosis or antagonism in the intestines of the human body. These microorganisms are beneficial bacteria such as Bifidobacterium, Lactobacillus and Streptococcus.

Defining probiotics as 'healthy germs when they consume modest amounts of living microbes' (FAO / WHO, 2001), prebiotics are the nutrients necessary for the growth of probiotics. Probiotics include Bifidobacterium, Clostridium, Lactobacillus, Bacillus, and Saccharomyces, which produce organic acids and antibiotics in the intestines of a representative bifidobacterium, Bifidobacterium, . This lactic acid bacterium is a representative probiotics, and various fermented foods, pastes, medicines and feed additives for livestock are being utilized variously. Recently, there have been many studies emphasizing the nutritional effects of lactic acid bacteria and various health functionalities of lactic acid bacteria in the intestinal tract.

In order to increase the in vivo activity of such a lactic acid bacterium, there is an increasing interest in synbiotics in which probiotics corresponding to live bacteria and prebiotics as nutrients thereof are mixed. However, the prebiotics used in the conventional synbiotics have a problem that digestion of digestive organs of the host progresses considerably and thus they do not act efficiently as a nutritional supply source of probiotics. Also, since the species specificity of prebiotics is insufficient, There is a problem in that it is not effective in increasing the activity.

On the other hand, yacon originates from the South American Andes, and contains various functional factors such as fructooligosaccharides and antioxidants in rootstock, which is similar to sweet potato. So far, researches on pharmacological effects and pharmacological values such as analysis of physicochemical components, amino acid analysis, changes in fructo-oligosaccharide content, anti-inflammatory activity, antioxidant and anticancer activities of extracts have been conducted. However, studies on the effect of yacon extracts on prebiotic and intestinal microbiological probiotics and the development of products utilizing them have been insufficient.

Therefore, the inventors of the present invention have confirmed the availability of yacon extract as synbiotic food together with probiotics as a prebiotic and completed the present invention.

Korean Registered Patent Publication B1 10-1381794

It is an object of the present invention to provide a synbiotics composition containing a yacon extract as a prebiotic and a microorganism belonging to the genus Bifidobacterium which specifically uses the yacon extract as a probiotics.

An object of the present invention is to provide a synbiotics food composition containing a yacon extract as a prebiotic and a microorganism belonging to the genus Bifidobacterium which specifically uses the yacon extract as a probiotics.

Another object of the present invention is to provide a synbiotics feed additive comprising a yacon extract as a prebiotic and a microorganism belonging to the genus Bifidobacterium which specifically uses the yacon extract as a probiotics.

The present invention provides a composition comprising a yacon extract and a microorganism belonging to the genus Bifidobacterium.

The yacon extract of the present invention means an extract extracted from yacon root. The extraction process of the present invention can be carried out by a conventional method known in the art. For example, water or an organic solvent is added to a yacon mass, and the mixture is subjected to reflux extraction, followed by filtration, followed by filtration and centrifugation to separate the residue and the filtrate. The separated filtrate is concentrated under reduced pressure to obtain a yacon extract. Preferably, the yacon troche is lyophilized prior to extraction.

The organic solvent which can be used in the present invention may be selected from ethanol, methanol, butanol, ether, ethyl acetate, chloroform or a mixed solvent of these organic solvents and water, preferably 80% ethanol. At this time, the extraction temperature is preferably 10 to 80 ° C, and the extraction time is preferably 6 to 24 hours. If the extraction temperature and the extraction time are exceeded, the extraction efficiency may be deteriorated or the component may be changed.

After obtaining the extract using a solvent, the extract can be obtained by cooling, heating and filtering at room temperature by a conventional method known in the art, or the solvent can be further evaporated, spray dried or freeze-dried.

The microorganism belonging to the genus Bifidobacterium of the present invention may include a microorganism belonging to the genus Bifidobacterium. Preferably, Bifidobacterium bifidum BGN4 , Bifidobacterium longum BORI and Bifidobacterium lactis , < / RTI >

The extract of yacon grass root of the present invention acts as a prebiotic and can be used more specifically for the growth of microorganisms of the genus Bifidobacterium among the microorganisms known as beneficial bacteria in the intestines. Therefore, it is possible to increase the microbial growth effect of the genus Bifidobacterium by including other microorganisms such as Bacillus subtilis and Bifidobacterium.

In the food composition of the present invention, the mixing ratio of the yacon extract and the Bifidobacterium genus microorganism may be 1: 2-5 by weight. Preferably, the weight ratio can be 1: 2-4.

The present invention provides a food composition comprising a yacon extract and a microorganism belonging to the genus Bifidobacterium.

In one embodiment, the food composition of the present invention may further comprise maltose, anhydrous crystalline glucose, indigestible maltodextrin, chicory extract, and vitamin C, depending on the type of food, in addition to the yacon extract and Bifidobacterium sp. But is not limited to.

The food composition of the present invention may have various oral formulations in a convenient form for ingestion and may have, for example, a granular powder, a liquid preparation, a gel or a solid preparation.

The food composition of the present invention may be provided in a general food or a health functional food. There is no particular limitation on the kind of the food. Examples of the foods to which the composition can be added include food products such as drinks, meat, sausage, bread, biscuits, rice cakes, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, Beverages, alcoholic beverages, and vitamin complexes, all of which include normal health foods.

The present invention also provides a feed additive comprising as a main ingredient a composition comprising a yacon extract and a microorganism belonging to the genus Bifidobacterium. The extraction method of the yacon extract of the present invention and the microorganism belonging to the genus Bifidobacterium are as described above.

The feed additive of the present invention is characterized in that it is added to a feed for livestock and fed to the livestock to increase the intestinal beneficial microflora, reduce the enterobacteriaceae, and increase the amount of the feed.

The present invention provides a synbiotics composition comprising a specific microorganism belonging to the genus Bifidobacterium that contains a yacon extract as a prebiotic and specifically utilizes the yacon extract as a probiotics, It can help the lactic acid bacteria to proliferate predominantly, thereby controlling the intestinal lactic acid bacteria environment.

Fig. 1 shows the kinds and content of sugar components contained in the ethanol extract of yacon mass.
FIG. 2 is a graph showing the results of measuring the number of bacteria by confirming the growth of three species of Bifidobacterium sp. In a medium containing a yacon trout extract and a medium not containing the extract.
FIG. 3 is a graph showing the results of measurement of the number of bacteria by confirming the proliferation of three Lactobacillus sp. Microorganisms in a medium containing yacon trout extract and a medium not containing it.

Hereinafter, the present invention will be described in more detail by way of examples. The contents not described in this specification can be sufficiently recognized and inferred by those of ordinary skill in the technical field or the like of the present invention, and the description thereof will be omitted.

≪ Example 1 > Preparation of yacon extract

Example 1-1 Preparation of Yacon

Yacon was collected in Korea and harvested on May 15, 2015 after it was planted on May 15, 2015 at the test packaging (560 m above sea level) of Chungbu-myeon, Pyeongchang-gun, Gangwon Province. Yacon used as a sample was cultivated in accordance with RDA standards. The samples were dried in a freezer (EYELA FDU-2100, EYELA Corp., Tokyo, Japan) for 5 days, and then placed in a mixer (Mini multi deluxe BL126DK blender, Tefal Co., Kaleo, France). Analysis of samples for each test item was expressed as live weight and repeated four times.

Example 1-2 Preparation of Ethanol Extract of Yacon

The extracts of Yacon Grassroot extract were prepared by adding 20-fold 80% ethanol to 10 g of lyophilized yacon roots and extracting them at 200 rpm for 24 hours at 25 ° C with a stirring extractor (VS-8480, Vision Scientific Co., After filtration through a filter paper (Whatman No. 2, Bukinghamshire, UK), the solvent was removed using a rotary vacuum concentrator (EYELA CCA-1110, Tokyo, Japan) and used as an extract. Each extract was diluted to 10 mL with extraction buffer and used in the experiment.

Example 2 Preparation of Bifidobacterium Microorganism Culture Medium

As the MRS-Control medium composition of the genus Bifidobacterium, 10 g of Dextrose, 10 g of Meat Peptone, 10 g of Beef extract, 5 g of yeast extract, 5 g of sodium acetate, 3 g of Disodium phosphate, 0.5 g of ammonium citrate, 0.1 g of magnesium sulfate, 0.05 g of maganese sulfate and 0.05% of L-cystrein hydrochloride were dissolved in 1,000 mL of distilled water and sterilized at 121 ° C for 15 minutes

(Table 1). As the MRS-Yacon medium composition, 1% yacon extract was added to the 1,000 mL control (MRS-Control) medium, and the mixture was subcultured three times or more before use in the experiment (Table 1).

 MRS-Control medium and MRS-yacon medium MRS-Control badge MRS-Yacon Badge Dextrose
Meat peptone
Beef extract
Yeast extract
Sodium acetate
Disodium phosphate
ammonium citrate
Tween 80
Magnesium sulfate
Manganese sulfate
L-cysteine hydrochloride
20.0 g
10.0 g
10.0 g
5.0g
5.0g
3.0 g
0.5 g
1.0 g
0.1 g
0.05 g
0.05%
Dextrose
Meat peptone
Beef extract
Yeast extract
Sodium acetate
Disodium phosphate
ammonium citrate
Tween 80
Magnesium sulfate
Manganese sulfate
L-cysteine hydrochloride
Yacon extract 20.0 g
10.0 g
10.0 g
5.0g
5.0g
3.0 g
0.5 g
1.0 g
0.1 g
0.05 g
0.05%
One%

   * Dissolve each treatment medium in 1,000 mL of distilled water and sterilize at 121 ℃ for 15 minutes.

< Example  3> Yacon Miniature  Components and content of extract

Fructooligosaccharide analysis was carried out according to the method of Food Code (KFDA, 2005). 50 ml of 80% ethanol was added to 10 g of the biological sample and the mixture was ground with a homogenizer (Ultra-Turrax T25 Basic Homogenizer, IKA Co., Staufen, Germany) at 9,500 rpm for 1 minute and filtered paper (No. 2, Whatman International, Maidstone, UK) and diluted to 50 mL. The extracted pretreatment solution was filtered through a membrane filter (0.45 μm, Milipore, Bedford, Mass., USA) and used for analysis. (1-kestose = GF2), nystose (GF3), and 1F-fructofuranosylnystose (GF4) for a standard solution preparation of fructooligosaccharides (Fructooligosaccharides Set, Wako, Osaka, Japan) Were weighed in 0.4 g each, placed in 20 mL volumetric flasks, and filled with distilled water to the markers. The HPLC (Waters 2695 HPLC, Waters Co., Milford, Mass., USA) analysis conditions were as follows: Column (Carbohydrate LC-NH2 Φ 4.6 × 250 mm 5 μm, Supercosil, Bellefonte, USA) (Waters 2414 Refractive Index Detector, Waters, Milford, MA, USA) at a flow rate of 1.0 mL / min and using acetonitrile: distilled water (70:30, v / v) Respectively. 10% fructooligosaccharide was contained.

Analysis of free sugar was carried out according to the method of Food Code (KFDA, 2005). 50 mL of 80% ethanol was added to 10 g of the biological sample and the mixture was ground for 1 minute at 9,500 rpm with a homogenizer (Ultra-Turrax T25 Basic Homogenizer, IKA Co., Staufen, Germany) , Whatman International, Maidstone, UK) and analyzed with an analyzer (Sugar analyzer, Toa Dkk, Tokyo, Japan). The analytical conditions were as follows: 0.2 M sodium hydroxide (NaOH) was used as an elution solvent, and a dedicated analytical column (PCI-520 Φ 4.6 × 150 mm 0.5 μm, Dkk Toa, Tokyo, Japan) was used. Standard solutions were prepared by dissolving 0.2, 0.4, 0.5, and 0.5 g of glucose, fructose, sucrose, and maltose in 50 mL of water, and diluting them to 100 mL with tertiary distilled water. 2 mL of the pretreated sample and standard solution was filtered through a membrane filter (0.45 μm, Milipore, Bedford, Mass., USA) and analyzed using an automatic injection device (Auto sampler ICA-200AS, Toa DKK, Tokyo, Japan) . At this time, the flow rate of the elution solvent was 0.7 mL / min.

As a result of analyzing the sugar content of yacon mass, the standard substance peak of the standard substance fructooligosaccharide and free sugar is as follows (Fig. 1). 7.6 g of fructo-oligosaccharide per 100 g of yacon body was contained, and sucrose was the most as 0.72 g of free sugars, followed by fructose 0.34 g and glucose 0.26 g (Table 1). Yacon contains fructose, glucose, sucrose, inulin, and fructooligosaccharides. Of these, fructooligosaccharides account for about 8% of the total.

 Yacon fructooligosaccharide, free sugar and reducing sugar content Artwork Fractooligosaccharides
(g / 100 g) Glucose
(g / 100 g) Fructose
(g / 100 g) Sucrose
(g / 100 g) Yacon 7.6 ± 0.03 ^z 0.26 ± 0.002 0.34 ± 0.005 0.72 ± 0.006

 zMean ± S.E.

Example 4 Influence of Yacon Extract on Microbial Growth of Bifidobacterium sp.

The Bifidobacterium Bifidobacterium BGN4 ( Bifidobacterium bifidum BGN4), Bifidobacterium ronggeom BORI (Bifidobacterium longum BORI ), Bifidobacterium lactis ) was used as a strain of Bifido Co, Ltd., Hongcheon, Korea. Three of the 2% Bifidobacterium sp. Microorganisms on the 1000 ml MRS medium were inoculated into each of the MRS media prepared in Example 2, incubated at 37 ° C for 18 hours, and then refrigerated at 4 ° C. These strains were dissolved in 1,000 mL of distilled water and sterilized at 121 ° C for 15 minutes in 0.05% L-cysteine hydrochloride. The growth curve of intestinal bifidus was obtained by inoculating the strains into the MRS-Control medium and the MRS-Yacon medium and measuring the number of viable cells per hour. The viable cell count was measured for 2 days at the end of 12 hours (0, 4, 8, 10, Respectively. The total number of bacteria was measured by using the method of Bifido Co., Ltd. in the medium using a yacon extract. That is, the total number of bacteria expressed using the MRS medium was counted, and the average value ± standard deviation was shown (Table 3 and FIG. 2).

culture
Time (h) B.bididum BGN4 B.longum BORI B.lactis   control   yacon root   control   yacon root   control   yacon root 0 2.20E + 07 2.50E + 07   4.40E + 07 3.60E + 07  4.12E + 07 5.90E + 07 4 1.30E + 08 1.70E + 08 3.03E + 08 2.83E + 08 4.40E + 08 6.14E + 08 8 1.00E + 09 1.10E + 09 9.55E + 08 9.25E + 08 1.73E + 09 2.17E + 09  10 1.20E + 09 1.30E + 09 1.01E + 09 1.33E + 09 1.88E + 09 2.20E + 09  11 1.30E + 09 1.40E + 09 1.08E + 09 2.00E + 09 1.90E + 09 2.23E + 09  12 1.40E + 09  1.50E + 09  1.10E + 09  2.35E + 09  2.00E + 09   2.23E + 09

As a result, it was confirmed that the proliferation of the three microorganisms of the genus Bifidobacterium inoculated into the medium containing the extract of yacon trout was improved compared to the control.

<Comparative Example 1> Lactobacillus effect of yacon extract on the Bacillus spp proliferative effect The present inventors same microbial growth and Examples 2 and 4 with respect to the Lactobacillus spp three kinds (Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus fermentum) (Table 4 and Figure 3).

culture
Time (h) L. acidophilus L. plantarum L.fermentum  control   yacon root  control   yacon root   control   yacon root   0 3.50E + 07   2.40E + 07 3.15E + 07 3.30E + 07 4.05E + 07 5.40E + 07   4 5.40E + 08 3.10E + 08 1.78E + 08 3.61E + 08 1.04E + 09 7.95E + 08   8 2.10E + 09 2.20E + 09 1.23E + 09 1.69E + 09 2.17E + 09 1.52E + 09   10 2.20E + 09 2.24E + 09 1.84E + 09 1.77E + 09 2.56E + 09 2.34E + 09   11 2.30E + 09 2.30E + 09 2.78E + 09 1.80E + 09 2.62E + 09 2.00E + 09   12 2.50E + 09 2.30E + 09 3.45E + 09 1.80E + 09 2.85E + 09 1.50E + 09

As a result, it was confirmed that the proliferation of the three Lactobacillus sp. Microorganisms inoculated on the medium containing the yacon trout extract was similar to that of the control group or inhibited the proliferation as time passed.

As a result, it was confirmed that the extract of Yacon Medicinalum had an effect of specifically proliferating microorganisms of the genus Bifidobacterium among the beneficial bacteria in the intestines. Therefore, by including the extract of Yacon grass root together with the microorganism of Bifidobacterium, it can be a synbiotic composition excellent in the effect of proliferating lactic acid bacteria.

Claims (7)

A synbiotic composition comprising a yacon extract and a Bifidobacterium longum BORI extracted from lyophilized yacon trout with ethanol. delete delete The method according to claim 1,
Wherein the composition is a mixture of yacon extract and Bifidobacterium longum BORI in a weight ratio of 1: 2 to 4: 1. 5. A synbiotic food composition comprising the composition of claim 1 or 4. The synbiotic food composition according to claim 5, wherein the food composition further comprises maltose, anhydrous crystalline glucose, indigestible maltodextrin, chicory extract, and vitamin C. A feed additive comprising the composition of claim 1 or 4.

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