KR102377028B1 - Leuconostoc mesenteroides subsp dextranicum NY203 strain, bitter taste-reduced dietary fiber prepared by using the same and functional food composition comprising the same - Google Patents

Abstract

The present invention relates to a leuconostoc mesenteroid subspecies dextranicum ( Leuconostoc mesenteroides subsp. dextranicum ) NY203 strain, a dietary fiber with reduced bitterness prepared using the same, and a health functional food composition comprising the same, wherein the dietary fiber comprises Bitterness is reduced and water absorption, swelling, fat absorption, cholesterol absorption, and cholic acid absorption are excellent, so it can be effectively used for diet or for the manufacture of antioxidant health functional foods.

Description

Leuconostoc mesenteroids subspecies dextranicum NY203 strain, a dietary fiber with reduced bitterness manufactured using the strain, and a health functional food composition comprising the same {Leuconostoc mesenteroides subsp dextranicum NY203 strain, bitter taste-reduced dietary fiber prepared by using the same and functional food composition comprising the same}

The present invention relates to a leuconostoc mesenteroids subsp. dextranicum NY203 strain, a dietary fiber with reduced bitterness prepared using the same, and a health functional food composition comprising the same, more particularly It relates to a health functional food composition comprising dietary fiber with reduced bitterness produced by culturing Leukonostok mesenteroid subspecies Dextranicum NY203 strain or a culture solution thereof using citron oil as a substrate.

Yuzu is the only special product in the southern coast of Korea, and it is estimated that about 20,000 tons of citron are produced annually around Geoje Island. Currently, citron production is over-produced, so farmers have almost given up on cultivation, and citron tea (for citron tea), which has been processed and produced so far, is consumed only in winter and is not stored during summer, so a huge amount is discarded. the current situation.

Citron is not only rich in vitamins C, P and E, but also contains limonin and nootropic compounds, which are currently found in grapefruit and oranges in the United States and Japan and are found to be powerful anticancer substances. It was confirmed that a fairly large amount of nomilin was contained. In particular, it was found that a large amount of compounds of the flavonoid system, such as carvone, coumarin, and terpinene, including limonene, which have high anticancer activity, were also found. Citron is a fruit that must be eaten with the peel because the peel contains a large amount of these physiologically active substances.

Up to now, citron has been mainly used as citron juice, and all it is to do is pickle the chopped citron in about 60% of sugar and then add a certain amount to water and use it as a tea. Citron juice sold by some companies recently is made by diluting citron juice by adding water to citron juice or by pressing citron and diluting it with water.

Looking at the registered patented technology for citron, the manufacturing method of citron tea (Korean Patent Registration No. 10-0026285), the manufacturing method of anchovy milk soup with citron and sugar added (Korean Patent Registration No. 10-0055045), and the use of citron Manufacturing method of traditional grain wine (Korean Patent Registration No. 10-0201532), Functional ice cream manufacturing method with citron added (Korean Patent Registration No. 10-0463077), Yuja Kimchi manufacturing method (Korean Patent Registration No. 10-0457133), etc. It can be seen that there is not much variety compared to other fruits.

The main components of the bitter taste of citron are naringin and hesperidin, which are contained in large amounts compared to other citrus fruits. Unlike other citrus fruits, citron has a bitter taste as it does not contain a naringinase enzyme that breaks down these bitter components, even when the fruit is ripe. It was impossible to juice the skin. In order to remove the bitter taste when manufacturing yuja syrup, it is made by pickling with sugar at least 60% of the total amount of fruit. Therefore, an excessive amount of sugar can mask the bitter taste of naringin, making it less bitter. Currently, most of the produced citron is consumed as citron juice, so excessive amounts of sugar are consumed.

The bitter taste present in some citrus fruits, especially citron, is present in excess, which limits the production of citron juice. Also, because of the strong bitter taste of ginseng, ginseng drinks are not becoming a favorite drink for the public, including children. Therefore, making a drink by removing the bitter taste of citrus fruits and ginseng is commercially very useful in popularizing citrus drinks or ginseng drinks.

In order to produce a naringinase enzyme that removes the bitter taste of citron, a method for preparing a fungal culture has been studied. Naringinase-producing strains are known to produce beta-glucosidase enzymes in addition to naringinase in culture. It is known that the mechanism by which the bitter taste of citron is removed is because beta-glucosidase and naringinase act step by step to decompose bitter naringin into non-bitter prunin and naringenin.

Therefore, the present inventors leuconostoc mesenteroid subspecies dextranicum ( Leuconostoc mesenteroides subsp. dextranicum ) NY203 strain and Aspergillus niger ( Aspergillus niger ) Culture of a strain or an enzyme derived therefrom using citron as a substrate It was confirmed that bitter taste was reduced in the resulting dietary fiber, water absorption power, swelling power, fat absorption power, cholesterol adsorption power and cholic acid adsorption power were excellent, and antioxidant activity appeared.

Accordingly, an object of the present invention is the leukonostok mesenteroid subspecies dextranicum NY203 strain deposited with accession number KCTC 13679BP, or a culture solution thereof; And Aspergillus niger strain, its culture medium, or to provide a method for producing dietary fiber comprising a culturing step of culturing at least one selected from the group consisting of fractions of the culture medium in the presence of a substrate.

Another object of the present invention is the leuconostok mesenteroid subspecies Dextranicum NY203 strain deposited with accession number KCTC 13679BP, or a culture solution thereof; And Aspergillus niger strain, its culture medium, or to provide a dietary supplement composition for diet comprising dietary fiber prepared by culturing at least one selected from the group consisting of fractions of the culture medium in the presence of a substrate.

Another object of the present invention is a leuconostok mesenteroid subspecies dextranicum NY203 strain deposited with accession number KCTC 13679BP, or a culture solution thereof; And Aspergillus niger strain, its culture medium, or to provide a health functional food composition for antioxidants comprising dietary fiber prepared by culturing at least one selected from the group consisting of fractions of the culture medium in the presence of a substrate.

Another object of the present invention is a leuconostok mesenteroid subspecies dextranicum NY203 strain deposited with accession number KCTC 13679BP, or a culture solution thereof; And Aspergillus niger strain, its culture medium, or one or more types of dietary fiber selected from the group consisting of fractions of the culture medium to reduce the bitter taste, induction of diet effect or to the use of enhancing antioxidant activity.

The present invention relates to a Leuconostoc mesenteroides subsp. dextranicum NY203 strain, a dietary fiber with reduced bitterness prepared using the same, and a health functional food composition comprising the same, according to the present invention Dietary fiber reduces bitterness, has excellent water absorption, swelling, fat absorption, cholesterol absorption, and cholic acid absorption, and exhibits antioxidant activity.

The present inventors confirmed that by culturing the strain using citron powder as a substrate, dietary fiber can be produced, and it can be usefully utilized in the manufacture of health functional food.

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

One aspect of the present invention is a leuconostok mesenteroid subspecies dextranicum NY203 strain deposited with accession number KCTC 13679BP, or a culture solution thereof; And Aspergillus niger ( Aspergillus niger ) It is a dietary fiber manufacturing method comprising a culturing step of culturing in the presence of a substrate at least one selected from the group consisting of a strain, its culture medium, or a fraction of its culture medium.

The culture medium contains the enzyme generated from the strain, and in the present specification, the supernatant of the strain culture medium containing the enzyme was used for manufacturing dietary fiber.

For example, in the examples of the present specification, the NY enzyme refers to the supernatant of the Leukonostok mesenteroid subspecies Dextranicum NY203 strain culture.

The concentration of the leukonostok mesenteroid subspecies dextranicum NY203 strain culture is 0.1 to 20.0% (v/v), 0.5 to 20.0% (v/v), 1.0 to 20.0% (v/v) or 5.0 to 20.0% (v/v), for example, may be 10.0 to 20.0% (v/v), but is not limited thereto.

The fraction of the culture medium is a result obtained by performing fractionation to separate a specific component or a specific component group from a culture medium containing various various components. In the present specification, the fraction of the culture medium is an enzyme isolated from the culture medium of the strain or enzymes thereof. It means some combination.

The fraction of the Aspergillus niger strain culture solution includes at least one first enzyme solution selected from the group consisting of pectinase, glucanase and arabinase; And it may be at least one selected from the group consisting of a second enzyme solution at least one selected from the group consisting of cellulase, hemicellulase and protease.

The concentration of the first enzyme solution is 0.01 to 2.00% (v/v), 0.05 to 2.00% (v/v), 0.10 to 2.00% (v/v), 0.50 to 2.00% (v/v), 1.00 to 2.00% (v/v), 0.01 to 1.00% (v/v), 0.05 to 1.00% (v/v), 0.10 to 1.00% (v/v) or 0.50 to 1.00% (v/v), for example, It may be 1.00% (v/v), but is not limited thereto.

The concentration of the second enzyme solution is 0.01 to 2.00% (v/v), 0.05 to 2.00% (v/v), 0.10 to 2.00% (v/v), 0.50 to 2.00% (v/v), 1.00 to 2.00% (v/v), 0.01 to 1.00% (v/v), 0.05 to 1.00% (v/v), 0.10 to 1.00% (v/v) or 0.50 to 1.00% (v/v), for example, It may be 1.00% (v/v), but is not limited thereto.

For example, the first enzyme solution in the present specification may be a UF enzyme, which contains pectinase, glucanase, and arabinase derived from Aspergillus niger strain. do.

According to one embodiment of the present invention, the UF enzyme exhibits an excellent dissolution rate for both soluble and insoluble dietary fiber.

For example, the second enzyme solution in the present specification may be a KN enzyme, which contains cellulase, hemicellulase and protease derived from Aspergillus niger strain.

According to one embodiment of the present invention, when the KN enzyme reacts with naringenin, glucose and rhamnose constituting bitter naringin, it can be decomposed into prunin and rhamnose without bitter taste, It can be used to reduce

According to one embodiment of the present invention, 1) leukonostok mecentroid subspecies Dextranicum NY203 strain or a culture solution thereof; and 2) Aspergillus niger strain, its culture solution or a fraction thereof, the dietary fiber has excellent water adsorption power, swelling power, fat adsorption power, cholesterol adsorption power and cholic acid adsorption power, so it has a diet effect and blood cholesterol lowering effect. It can be used for induction purposes.

The substrate may be a vegetable fiber, and the vegetable fiber may be derived from at least one selected from the group consisting of fruits, leaves, stems, and roots of plants, but is not limited thereto.

The concentration of vegetable fiber is 1 to 20% (w / v), 2.5 to 20% (w / v), 5 to 20% (w / v), 10 to 20% (w / v), 1 to 10% ( w/v), 2.5 to 10% (w/v) or 5 to 10% (w/v), for example, may be 10% (w/v), but is not limited thereto.

The incubation step is 1 to 15 hours, 1 to 12 hours, 1 to 9 hours, 2 to 15 hours, 2 to 12 hours, 2 to 9 hours, 3 to 15 hours, 3 to 12 hours, 3 to 9 hours, 6 to 15 hours or 6 to 12 hours, for example, may be performed for 6 to 9 hours, but is not limited thereto.

Another aspect of the present invention is a leuconostok mesenteroid subspecies dextranicum NY203 strain deposited with accession number KCTC 13679BP, or a culture solution thereof; And Aspergillus niger strain, its culture medium, or a dietary fiber prepared by culturing at least one selected from the group consisting of fractions of its culture medium in the presence of a substrate is a dietary supplement composition for diet.

The concentration of the leukonostok mesenteroid subspecies dextranicum NY203 strain culture is 0.1 to 20.0% (v/v), 0.5 to 20.0% (v/v), 1.0 to 20.0% (v/v) or 5.0 to 20.0% (v/v), for example, may be 10.0 to 20.0% (v/v), but is not limited thereto.

The fraction of the Aspergillus niger strain culture solution includes at least one first enzyme solution selected from the group consisting of pectinase, glucanase and arabinase; And it may be at least one selected from the group consisting of a second enzyme solution at least one selected from the group consisting of cellulase, hemicellulase and protease.

The substrate may be a vegetable fiber, and the vegetable fiber may be derived from at least one selected from the group consisting of fruits, leaves, stems, and roots of plants, but is not limited thereto.

The concentration of vegetable fiber is 1 to 20% (w / v), 2.5 to 20% (w / v), 5 to 20% (w / v), 10 to 20% (w / v), 1 to 10% ( w/v), 2.5 to 10% (w/v) or 5 to 10% (w/v), for example, may be 10% (w/v), but is not limited thereto.

Another aspect of the present invention is a leuconostok mesenteroid subspecies dextranicum NY203 strain deposited with accession number KCTC 13679BP, or a culture solution thereof; And Aspergillus niger strain, its culture medium, or a dietary fiber prepared by culturing at least one selected from the group consisting of fractions of the culture medium in the presence of a substrate is a health functional food composition for antioxidants.

The concentration of the leukonostok mesenteroid subspecies dextranicum NY203 strain culture is 0.1 to 20.0% (v/v), 0.5 to 20.0% (v/v), 1.0 to 20.0% (v/v) or 5.0 to 20.0% (v/v), for example, may be 10.0 to 20.0% (v/v), but is not limited thereto.

The fraction of the Aspergillus niger strain culture solution includes at least one first enzyme solution selected from the group consisting of pectinase, glucanase and arabinase; And it may be at least one selected from the group consisting of a second enzyme solution at least one selected from the group consisting of cellulase, hemicellulase and protease.

The substrate may be a vegetable fiber, and the vegetable fiber may be derived from at least one selected from the group consisting of fruits, leaves, stems, and roots of plants, but is not limited thereto.

The concentration of vegetable fiber is 1 to 20% (w / v), 2.5 to 20% (w / v), 5 to 20% (w / v), 10 to 20% (w / v), 1 to 10% ( w/v), 2.5 to 10% (w/v) or 5 to 10% (w/v), for example, may be 10% (w/v), but is not limited thereto.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. In general, in the production of food or beverage, the health functional food composition of the present invention may be added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material.

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all foods in a conventional sense.

The beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. As the above-mentioned natural carbohydrates, monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame may be used. . The ratio of the natural carbohydrate may be appropriately determined by the selection of those skilled in the art.

In addition to the above, the health functional food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin , alcohol, carbonation agent used in carbonated beverages, and the like. In addition, the health functional food composition of the present invention may contain fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives may also be appropriately selected by those skilled in the art.

The present invention relates to a leuconostoc mesenteroid subspecies dextranicum ( Leuconostoc mesenteroides subsp. dextranicum ) NY203 strain, a dietary fiber with reduced bitterness prepared using the same, and a health functional food composition comprising the same, wherein the dietary fiber comprises Bitterness is reduced, water absorption power, swelling power, fat adsorption power, cholesterol adsorption power, cholic acid adsorption power and antioxidant activity are excellent, so it can be effectively used in the manufacture of dietary or antioxidant health functional food.

1 is a photograph of the cellulose or starch degradation results of 200 isolated strains of the present invention.
Figure 2 is a photograph of the results of selection of strains for decomposing 13 kinds of starch and cellulose.
3 is a photograph showing the results of selection of the best polysaccharide degrading strain.
Figure 4a is a photograph showing the dissolution rate of yujabak dietary fiber using an isolated strain.
Figure 4b is a graph showing the dissolution rate of yujabak dietary fiber using the isolated strain.
Figure 5a is a photograph showing the dissolution rate of yujabak dietary fiber using a commercial enzyme.
Figure 5b is a graph showing the dissolution rate of yujabak dietary fiber using a commercial enzyme.
6A is a photograph showing the removal rate of bitter components of yujabak dietary fiber using a commercial enzyme.
Figure 6b is a graph showing the removal rate of bitter components of yujabak dietary fiber using a commercial enzyme.
7A is a photograph showing the dissolution rate and bitter taste component content of yujabak dietary fiber according to the substrate concentration.
7b is a photograph showing the dissolution rate and bitter taste component content of yujabak dietary fiber according to the enzyme concentration.
7c is a photograph showing the dissolution rate and bitterness component content of yujabak dietary fiber according to the reaction time.
Figure 8a is a graph showing the dissolution rate of yujabak dietary fiber according to the substrate concentration.
Figure 8b is a graph showing the dissolution rate of yujabak dietary fiber according to the enzyme concentration.
Figure 8c is a graph showing the dissolution rate of yujabak dietary fiber according to the reaction time.
9A is a graph showing the bitter taste component content of yujabak dietary fiber according to the substrate concentration.
9B is a graph showing the bitter taste component content of yujabak dietary fiber according to the enzyme concentration.
9c is a graph showing the bitter taste component content of yujabak dietary fiber according to the reaction time.
10 is a photograph showing the TLC results of the mixed enzyme-treated yujabak dietary fiber.
11A is a photograph showing the upper and lower layers and powder of yujabak dietary fiber treated with mixed enzyme.
Figure 11b is a graph showing the dissolution rate of the mixed enzyme-treated yujabak dietary fiber.
12a is a result of HPLC analysis of free sugars of yujabak powder treated with mixed enzymes.
12b is a result of analysis of HPLC free sugar of yujabak powder treated with mixed enzymes for each sample.
13 is a graph showing the distribution of dietary fiber of the enzyme-treated citron powder.
14 is a scanning electron microscope (SEM) photograph of the enzyme-treated citron powder.
15 is a graph showing the results of Fourier transform infra-red spectroscopy (FTIR) analysis of yujabak dietary fiber powder treated with mixed enzyme.
16 is an HPLC analysis graph showing changes in functional components of yujabak dietary fiber powder treated with mixed enzymes.
Figure 17a is the result of investigating the degree of bitterness of the enzyme-treated yujabak dietary fiber powder by an electrophysiological technique.
17B is a graph showing the results of investigation of the degree of bitterness of enzyme-treated yujabak dietary fiber powder by an electrophysiological technique.
Figure 18a is a graph showing the water absorption capacity of the enzyme-treated yujabak dietary fiber powder.
Figure 18b is a graph showing the expansion force of the enzyme-treated yujabak dietary fiber powder.
Figure 18c is a graph showing the liposuction power of the enzyme-treated yujabak dietary fiber powder.
18D is a graph showing the cholesterol adsorption capacity of the enzyme-treated yujabak dietary fiber powder.
Figure 18e is a graph showing the expansion force of the enzyme-treated yujabak dietary fiber powder.

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

Throughout this specification, "%" used to indicate the concentration of a specific substance is (weight/weight)% solid/solid, (weight/volume)%, and (weight/volume)% for solid/solid, and Liquid/liquid is (vol/vol) %.

All experimental results were measured three times and expressed as the mean and standard deviation, and analysis of variance (ANOVA) was performed to test the significance between the mean values for each treatment using the SAS program (Package relwase 8.01). For items with significant differences between the means, a significance test was performed at the p<0.05 level using Duncan's multiple range test.

Example 1: Selection of enzymes for manufacturing functional dietary fiber from citron

1-1. Strain culture and selection

As traditional foods used in this experiment, about 200 kinds of soybean paste, red pepper paste, kimchi, and makgeolli were collected from all over Jeollanam-do. Reagents used for the analysis were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).

After suspending 20 g of the collected sample in 80 mL of 0.85% (w/v) sterile physiological saline, MRS agar (Difco, Sparks, MD, USA), LB agar (Difco), YM agar as a nutrient medium It was spread on the medium and incubated at 37° C. for 48 to 96 hours. After culturing, colonies showing the properties of lactic acid bacteria were selected (picking). As shown in FIG. 1, about 200 selected colonies were purely separated in the same medium and subcultured for three generations, then a single colony was taken and cultured in MRS liquid medium.

MRS agar (Difco, Sparks, MD, USA) added with 1% CMC (sodium carboxymethyl cellulose) to check the dietary fiber decomposition ability and 1% starch to check the starch decomposition ability, LB agar (Difco), YM After adding 5 mL of agar (Difco) to a 6-well microplate, incubate in MRS liquid medium, smear 10 uL each showing 0.5 at OD 600 nm, incubate at 37°C for 20 hours, and then 0.1% After staining with Congo red for 15 minutes, the size of the decomposed cellulose ring was confirmed by decolorization with 1M NaCl.

In addition, after 20 minutes of staining with iodine, iodine was removed to confirm the size of the decomposed starch ring. At this time, the activity of the selected bacteria is divided into 0 mm (-), 0 to 2 mm (+), 2 to 3 mm (++), and 3 mm or more (+++) depending on the size of the ring, so that the resolution of cellulose or starch is improved. 13 excellent strains were selected and shown in FIG. 2 . Each bacteria was cultured in LB or MRS liquid medium, suspended in a 50% glycerol solution, and stored at -80° C. for use in the experiment.

1-2. Gene PCR screening and identification for polysaccharide digestion

The 16S rRNA gene sequences of NY1, 24, 74, 104, 126, 128, 154, 155, 158, 160, 174, 184 and 203 of 13 strains selected in 1-1 were analyzed. Among them, only 8 strains, 74, 104, 126, 154, 155, 174, 184, and 203 strains, could be identified.

Specifically, after DNA was extracted from the cells, the 16S rRNA gene was amplified by PCR using universal primers, 27F and 1492R. The amplified gene product was requested by Macrogen, and the nucleotide sequence was deciphered by using primers for universal sequencing, 785F and 907R primers. The primers used are shown in Table 1 below.

Primers for PCR and sequencing for identification of 8 strains for polysaccharide degradation SEQ ID NO: designation Sequence (5'-3') One 785 F GGA TTA GAT ACC CTG GTA 2 907 R CCG TCA ATT CMT TTR AGT TT 3 27 F AGA GTT TGA TCM TGG CTC AG 4 1492 R TAC GGY TAC CTT GTT ACG ACT T

The 16S rRNA gene sequences of 5 strains were compared using the NCBI nucleotide sequence database (www.ncbi.nlm.nih.gov./blast/) by BLAST search.

As shown in Table 2, 8 types of 16S rRNA gene nucleotide sequences are respectively Panibacillus yonginensis ( Paenibacillus yonginensis ) (74), Leuconostoc mesenteroids subspecies dextranicum ( Leuconostoc mesenteroides subsp. dextranicum ) (104, 154, 155, 174, 184, 203), and Leuconostoc citreum ( Leuconostoc citreum ) (126) were confirmed to have more than 95% homology.

Identification results of 8 strains for polysaccharide degradation Strain no. Accession no. Identification (%) Identity (%) food raw material
Item number 74 CP_014167.1 Paenibacillus yonginensis 95 Not applicable 104 CP_012009.1 Leuconostoc mesenteroides subsp. dextranicum 99 A is 004900 126 LC_096222.1 Leuconostoc citreum 100 A is 004800 154 CP_012009.1 Leuconostoc mesenteroides subsp. dextranicum 99 A is 004900 155 CP_012009.1 Leuconostoc mesenteroides subsp. dextranicum 99 A is 004900 174 CP_012009.1 Leuconostoc mesenteroides subsp. dextranicum 99 A is 004900 184 CP_012009.1 Leuconostoc mesenteroides subsp. dextranicum 99 A is 004900 203 CP_012009.1 Leuconostoc mesenteroides subsp. dextranicum 99 A is 004300

1-3. Cellulose or starch resolution (cellulase) TLC screening

In order to screen the strains, eight strain cultures were confirmed by thin layer chromatography (TLC) analysis.

Specifically, first, the bacteria were grown in LB medium to which 1% soluble starch or 1% cellulose (CMC) was added at 37° C. for 3 days until OD 2.0, and then the primary bacterial culture medium was OD = The concentration of bacteria was titrated to 0.5. 5 kDa cutoff ultrafiltration (Pall, Port Washington, NY, USA) of the cultured LB medium to remove polysaccharides in the medium to confirm the polysaccharide resolution, 1% soluble starch or 1% CMC is added did

The isolated strain was cultured at 37° C. for 72 hours, then centrifuged at 8,000 rpm for 10 minutes (6200, Kubota, Tokyo, Japan), and the supernatant was used as a TLC screening sample. Reaction conditions were 0.2M Na-acetate, pH 5.2, 1% soluble starch or 1% cellulose, and the enzyme-containing strain supernatant was mixed and reacted at 37°C for 24 hours. 1 uL of the reaction solution was added dropwise and developed with a developing solvent (nitromethane: 1-propanol: water=2: 5: 1.5 (v/v/v)), UV 245 nm and a coloring solvent (0.5% naphtal, 5% H ₂ SO ₄ in ethanol), and then baked in an oven at 105° C. for 3 minutes to identify spots.

To select a strain with excellent polysaccharide resolution among 8 strains, FS (0.1% fructose, sucrose), GR (0.1% glucose, rahmnose), IMO (1%, Isomaltose series), and MO (1% Maltose series) were used. 10% of the strain supernatant (No. 74, 104, 126, 154, 155, 174, 184, 203) was added to 0.05 M sodium acetate buffer (Na- acetate), pH 5.2, reacted at 37° C. for 6 hours, and analyzed by thin layer chromatography (TLC). Citron powder used as a substrate is a conventional citron variety harvested in November, and it is a powder obtained by freeze-drying and grinding the juiced, seed-removed citron.

As can be seen in FIGS. 3, 4a, 4b and Table 3, as a result of investigating the dissolution rate using No. 126, 174, and 203 as superior strains among the strains selected for the mammary gland, the dissolution rate of dietary fiber was 47.4%, 39.4%, and 62.0% in the case of No. 203 As a result, strain 203 had the highest efficiency.

NY126 NY174 NY203 Dietary fiber dissolution rate (%) 47.4b 39.4c 62.0a

1-4. Securing the optimal strain for solubilization of water-soluble dietary fiber in citron

Using 11 commercial enzymes shown in Table 4 below, 10% of yujabak substrate was added and reacted for 6 hours.

Strains and enzymes for dissolving commercial soluble dietary fiber and removing bitter components product derived bacteria activation One PT Aspergillus aculeatus Polygalactronase 2 VS Aspergillus aculeatus β-Glucanase, Arabanase, Cellulase,
Hemicellulase, Xylanase 3 CL Trichodermareesei ß-glucanase, Cellulase 4 CP Aspergillus niger Pectinase, Polygalacturonase,
Pectinmethylesterase 5 PR Aspergillus niger Pectinase, Pectinmethylesterase 6 UF Aspergillus niger Pectinase, Glucanase, Arabinase 7 TA Aspergillus oryzae Tannin acylhydrolase 8 BG Trichodermareesei ß-glucanase, Cellulose 9 PL Aspergillus niger β-Glucosidase, Cellobiase 10 AC Aspergillus niger β-glucosidase, cellulase, hemicellulose 11 KN Aspergillus niger Cellulase, Hemicellulase, Protease

As can be seen in Figures 5a and 5b, all the enzymes showed a dissolution rate of 13.2 to 51.9% lower than the 62.0% dissolution rate shown in NY203.

Among them, the highest solubility is UF enzyme 6 (PECLYVE UF CLEANER, Glucanase 50 U/g, Arabinase 450U/g, Lyven Zac Normmandial), Aspergillus niger ( Aspergillus niger ) It was performed by selecting an enzyme solution containing derived pectinase, glucanase and arabinose. The reaction conditions were 0.05 M sodium acetate buffer, pH 5.2, and 10% yujabak was added as a substrate, followed by reaction in 1% enzyme solution at 50° C. for 6 hours.

1-5. Securing the optimal strain (enzyme) for removing the bitter taste of yujabak

The bitter taste is mainly due to glycosides (glucose and rhamnose) bound to aglycone, and it is known that most of the bitter taste disappears when the sugar component of this glycoside is decomposed into an aglycone state during fermentation. It was attempted to remove bitterness by decomposing sugars such as rhamnose and glucose, which are bound to naringin, etc., which have a characteristic bitter taste of citron, into non-glycosides using beta-glucosidase and nariniginase.

Using 1% of 11 commercial enzyme solutions, 10% yujabak was added as a substrate in 0.05 M sodium acetate buffer, pH 5.2, and reacted for 6 hours.

As can be seen in FIGS. 6a and 6b, the rhamnose sugar content after the enzymatic reaction varied from 0.48 to 3.1 mg/mL, and among them, KN enzyme 11 (Celluase KN, B-glucosidase 20 U/g Vision Biocam) ), Aspergillus niger In the case of an enzyme solution containing derived cellulase, hemicellulase, and protease, the decomposition rate of naringin was the highest and the derived rhamnose sugar content was 3.1 mg/mL, which was the highest.

Example 2: Optimization of Enzyme for Production of Functional Dietary Fiber Using Yuja Bak

Leukonostok mesenteroid subspecies dextranicum Using the supernatant (10%) of the NY203 strain culture, the dissolution rate of insoluble dietary fibers, cellulose and hemicellulose, Aspergillus niger The dissolution rate of pectin, a water-soluble dietary fiber, using UF (1%), Aspergillus niger The degree of removal of bitter taste components was investigated using KN (1%). Basically, the reaction was carried out under the conditions of 0.05 M sodium acetate buffer at pH 5.2.

Basically, the substrate, yujabak, was 10%, the enzyme was 10% NY, 1% commercial UF, 1% commercial KN, and the reaction time was 6 hours for NY, and 3 to 6 hours for UF and KN, respectively. The concentration of yujabak as a substrate is 0.0, 1.0%, 2.5%, 5.0%, 10.0% and 20.0%, and the concentration of the enzyme is 0.00, 0.01%, 0.10%, 0.50%, 1.00%, 2.00%, 5.00%, 10.00 % and 20.00%, and the reaction time was applied up to 0.0, 1.0, 1.5, 2.0, 3.0, 4.0, 5.0, 6.0, 9.0, 12.0 and 15.0 h. (GP (0.1% glucose, pectin), FS (0.1% fructose, sucrose), Isomalto series (IMO), Maltose series (MO)) The solubilization rate was determined by comparing the weight difference between samples before and after enzyme treatment. is expressed in %.

As can be seen in FIGS. 7, 8 and Tables 5 to 7, the dissolution rate of dietary fiber according to the concentration of the yujabak substrate used as a substrate is 10% for the NY enzyme and 20% for the UF enzyme, but the solubilization of the enzyme and the substrate For efficiency, it can be seen that the content of 10% is optimal. As for the dissolution rate of dietary fiber by enzyme concentration, it was found that 10% of NY enzyme and 1% of UF enzyme were optimal. As for the dissolution rate by reaction time, 6 hours for the NY enzyme and 3 hours for the UF enzyme were found to be optimal. In general, the NY enzyme had a higher solubilization rate of dietary fiber than the commercial UF.

Dietary fiber dissolution rate and bitter taste component content by yujabak substrate concentration Concentration of substrate (%) 0.0 1.0 2.5 5.0 10.0 20.0 NY 10%
Dietary Fiber
Dissolution rate (%) - 29.2 ^b 38.4 ^ab 40.1 ^ab 49.2 ^a 31.9 ^b UF 1% dietary fiber
Dissolution rate (%) - 3.9 ^d 9.6 ^d 20.9 ^c 41.2 ^b 48.2 ^a KN 1% rhamnose (%) - 0.1 ^e 0.3 ^d 1.3 ^c 2.0 ^b 2.2 ^a

Dietary fiber dissolution rate and bitter taste component content by enzyme concentration Enzyme concentration (%) 0.00 0.01 0.10 0.50 1.00 2.00 5.00 10.00 20.00 NY
Dietary Fiber
Dissolution rate (%) 0 ^d - 12.5 ^c 14.5 ^c 17.5 ^c - 28.1 ^b 42.4 ^a 47.0 ^a UF dietary fiber
Dissolution rate (%) 0 ^d 3.2 ^d 9.7 ^c 15.9 ^b 28.6 ^a 31.3 ^a - - - KN
Naringin (%) 100.0 ^a 91.4 ^b 38.5 ^c 34.0 ^d 24.9 ^e 20.4 ^f - - - KN
Rhamnose (%) 0.0 ^d 0.0 ^d 50.2 ^c 77.1 ^b 90.6 ^a 90.6 ^a - - -

Dietary fiber dissolution rate and bitter taste component content by reaction time Reaction time (h) NY
Dietary Fiber
Dissolution rate (%) UF
Dietary Fiber
Dissolution rate (%) KN
Naringin (%) KN
Rhamnose (%) 0.0 0 0 100.0 ^a 0.0 ^g 1.0 7.3 ^f 1.2 ^e 78.6 ^b 27.9 ^f 1.5 - 10.4 ^d 71.4 ^c 37.1 ^e 2.0 10.4 ^e 23.2 ^c 57.1 ^d 55.7 ^d 3.0 23.2 ^d 40.3 ^b 50.0 ^e 65.0 ^c 4.0 - 39.5 ^b - - 5.0 - 41.2 ^ab - - 6.0 42.4 ^bc 43.2 ^a 42.9 ^f 84.3 ^a 9.0 44.8 ^ab - - - 12.0 40.3 ^c - 28.6 ^g 78.6 ^b 15.0 45.2 ^a - - -

Naringin, which produces bitter taste, has a structure consisting of naringenin + glucose + rhamnose, and is decomposed into pruning (Naringenin glucoside) and rhamnose without bitter taste by KN enzyme treatment.

As can be seen in Fig. 9, Tables 5, 6 and 7, the content of naringin, which is a bitter component, decreased, and the amount of rhamnose that was removed by the enzymatic action increased. The removal rate of bitter components by concentration of yujabak substrate used as a substrate is highest when the KN enzyme is 2%, but 1% content is optimal for the solubilization efficiency of the enzyme and substrate. The removal rate of the bitter taste component by enzyme concentration was found to be optimal when the KN enzyme was 1 to 2%, and the bitter taste component removal rate by reaction time was found to be optimal for 3 to 6 hours when the reaction was performed with the KN enzyme.

Example 3: Complex reaction of three enzymes for the production of functional dietary fiber using citron

The concentration of each enzyme in the yujabak dietary fiber substrate was confirmed by TLC analysis when each or a combination of NY 10%, UF 1%, and KN 1% was applied. In the presence of 0.05 M sodium acetate buffer, citron powder was added as a 10% substrate at pH 5.2 and reacted for 6 hours. Here, it means R (0.1% Rhamnose), GG (0.1% Glucose, Galacturonic acid (pectin precursor), FS (0.1% fructose, sucrose), Isomalto series (IMO), Maltose series (MO).

As can be seen in FIG. 10, the NY enzyme decomposed insoluble cellulose, the UF enzyme decomposed pectin, a soluble dietary fiber, and the KN enzyme decomposed bitter naringin, and it can be seen that the content of rhamnose liberated therefrom is increased.

Example 4: Quantitative and qualitative analysis of mixed enzyme-treated dietary fiber

4-1. Powder and dissolution rate of enzyme-treated citron meal fiber

The concentrations of each enzyme in the yujabak dietary fiber substrate were NY 10%, UF 1%, and KN 1%, respectively or sequentially, and then the upper and lower layers, powder form and dissolution rate were analyzed. In the presence of 0.05 M sodium acetate buffer, citron powder was added as a 10% substrate at pH 5.2 and reacted for 6 hours.

As can be seen in FIG. 11a , the powder of citron meal fiber that was lyophilized after sequentially reacting 10% NY + 1% UF + 1% KN was decomposed into pellets (pellet) as insoluble and soluble dietary fiber compared to the untreated case. ) was found to be significantly reduced.

As can be seen in FIG. 11b and Table 8, the dissolution rate was the highest in the NY+UF+KN mixed enzyme-treated powder.

unprocessed NY UF NY+UF NY+UF+KN Dissolution rate (%) 2.5 ^d 48.5 ^bc 42.1 ^c 50.2 ^b 67.2 ^a

4-2. Investigation of free sugar content of enzyme-treated citron powder

The free sugar content of the mixed enzyme-treated citron powder was investigated. Cellulose is liberated from glucose, hemicellulose from arabinose, and pectin from galacturonic acid. After passing through the membrane filter), the increase in their content was investigated using HPLC.

Specifically, HPLC was performed using an Agilent 1216 infinity LC series system (Agilent Technologies, Palo Alto, CA, USA). The analytical column was ZORBAX eclipse plus C18 (4.6x250mm, 5-Micron, Agilent Technologies, Palo Alto, CA, USA), and the solvent composition was A:0.1% formic acid, B:methanol-acetonitrile was used. did The solvent gradient starts with A:80, B:20, A:60, B:40 for 5-10 minutes, A:50, B:50 for 10.1-15 minutes, A:30, B for 15.1-20 minutes :70, A:0, B:100 for 20.1-25 minutes, A:80, B:20 for 25.1-30 minutes were analyzed. The solvent flow rate was 0.5 mL/min, the column temperature was fixed at 35 °C, 10 uL of the sample was injected, and detection was performed at 280 nm.

As can be seen in FIGS. 12a, 12b and Table 9, glucose, fructose and arabinose were increased in the NY enzyme-treated powder compared to the untreated powder, and the UF enzyme-treated powder showed little change in glucose and fructose. The content of pectin was significantly increased. In the case of NY+UF enzyme treatment, the contents of glucose and pectin increased, but the difference was insignificant compared to the case of single enzyme treatment. In the case of NY + UF + KN final mixed enzyme treatment, it was confirmed that all of pectin, glucose, fructose and arabinose were the highest.

content(%) unprocessed NY UF NY+UF NY+UF+KN Galacturonic Acid (Pectin) - 0.09 ^d 0.89 ^b 0.81 ^c 0.96 ^a glucose 1.10 ^d 1.94 ^a 1.28 ^c 1.71 ^b 2.03 ^a fructose 1.37 ^b 1.55 ^a 1.40 ^ab 1.33 ^b 1.49 ^ab arabinos - 0.29 ^c 0.38 ^b 0.37 ^b 0.41 ^a

4-3. Investigation of dietary fiber composition of enzyme-treated citron powder

The insoluble and soluble dietary fiber content of the citron powder enzyme-treated powder was measured by the enzymatic gravimetric method of Prosky et al. Accurately weigh 1 g of each dry sample using citron meal fiber, add 50 mL of phosphate buffer (pH 6.0), and then add amylase, protease, and amyloglucosidase. was sequentially hydrolyzed using Thereafter, the dietary fiber content was calculated through the difference between the measured values of each protein and ash content.

In order to derive the insoluble dietary fiber (IDF) content, the difference between the measured protein and ash content of the residue washed with 95% and 78% ethanol and acetone after enzyme treatment is washed with acetone and then oven-dried is used. to perform the calculation. In addition, after enzyme treatment, 95% ethanol at 60°C was added to the filtrate to dry and measure the precipitated solid, and the soluble dietary fiber (SDF) content was calculated using the difference between the measured values of protein and ash content, and the insoluble And the content of total dietary fiber (TDF) was calculated using the sum of the content of soluble dietary fiber.

As can be seen in Table 10, the dietary fiber content was reduced and the NY enzyme-treated powder significantly reduced insoluble dietary fiber due to decomposition of cellulose and hemicellulose. In the UF enzyme-treated powder, soluble dietary fiber decreased due to the dissolution of pectin. In the case of NY+UF+KN enzyme-treated powder, both soluble and insoluble dietary fiber compositions were dissolved.

TDF (%) IDF(%) SDF (%) unprocessed 34.52 ^a 17.85 ^a 16.67 ^a NY processing 18.06 ^b 7.22 ^c 10.84 ^b UF processing 18.04 ^b 11.68 ^b 6.36 ^c NY+UF processing 9.51 ^c 7.03 ^c 2.48 ^d NY+UF+KN processing 8.76 ^c 6.09 ^c 2.67 ^d

As can be seen in FIG. 13, in particular, compared to the untreated powder, in the case of the NY enzyme-treated powder, the cellulose decreased by 2 times and the hemicellulose decreased by 3 times, and in the case of the UF enzyme-treated powder, the cellulose decreased by 2 times, and the pectin was 5 times It was confirmed that the UF enzyme showed a high dissolution rate for both soluble and insoluble dietary fiber. In the case of NY+UF+KN enzyme-treated powder, it was confirmed that 60 to 65% of the total was dissolved, which was consistent with the dissolution rate result.

Example 5: Confirmation of changes in structure and chemical properties of enzyme-treated yujabak dietary fiber

Structural changes of untreated, NY, UF and mixed enzyme-treated powders thereof were confirmed using a Scanning Electron Microscope (SEM, ZEMINI SEM, FEI Co., The Netherlands). The image of the yujabak dietary fiber particles was irradiated at 400 times magnification.

As can be seen in Figure 14, Compared to the untreated powder, the NY + UF + KN enzyme-treated powder shows a collapse of the structure, and it can be observed that there are many voids in the tissue, and the tissue is inflated like a balloon to form a structure that is easy to dissolve.

FTIR analysis (Fourier transform infra-red spectroscopy, perkin Elmer Spectrum 400) was performed to confirm the change in the chemical properties of the enzyme-treated dietary fiber.

As a result of the analysis, the dietary fiber treated with yujabak mixed enzyme was 1) 3500 to 2900 cm ^-1 , 2) 1700 to 1300 cm ^-1 , 3) 1200 to 900 cm in the FTIR spectrum. It is marked as ^-1 . According to the previous research results, 1) means OH parasitic formation in each range, and mainly water or ethanol characteristics are shown. 2) mainly means the presence, increase or generation of phenolic compounds, and represents the main characteristics of flavonoids and phenols. 3) indicates the expansion or increase of the carbonyl group, and an increase in the curve height means an increase in the C=O bond.

As can be seen in FIG. 15, the peak patterns of all untreated, NY, and UF powders are similar, and most of the peaks are concentrated at 1700 to 1300 cm ^-1 indicating the presence of phenolic compounds as flavonoids. In particular, the final NY+UF+KN mixed enzyme-treated powder has a larger peak size compared to the untreated powder or single or mixed enzyme-treated powder, and creates aromatic rings through the presence of OH groups and the combination of phenolic compounds. showed an increase in this.

Example 6: Measurement of functional component change and bitter taste of mixed enzyme-treated dietary fiber

HPLC analysis was performed by comparing the untreated sample and the NY+UF+KN mixed enzyme-treated sample in order to characterize the change in the functional components of the enzyme-treated dietary fiber.

As can be seen in FIGS. 16 and 11, in the untreated final powder, naringin and neohesperidin, which are known to give bitter taste, were slightly decreased while the diet components such as narirutin and hesperidin were slightly decreased. It was found that the bitter taste was found to be reduced by 50 to 65% or more.

Changes in functional component composition of mixed enzyme-treated citron dietary fiber powder (mg/g DW) nariloutine Naringin hesperidin neohesperidin unprocessed 17.94 5.49 10.85 4.75 NY+UF+KN 14.99 2.74 8.81 2.15

In other words, naringin and hesperidin are glycosides bound to sugar, so they taste bitter, but when the sugars bound by beta-glucosidase and naringinase are decomposed during fermentation and converted to the aglycones naringenin and hesperetin, the bitter taste disappears. .

The degree of bitterness was measured by treating frog eggs with bitter taste receptor-expressed single and mixed enzyme-treated dietary fiber powder at a concentration of 10 ug/mL.

Specifically, mature and healthy clawed frog (Xenopus) oocytes of stage 5 or 6 were surgically collected from the ovaries and then obtained by treatment with collagenase. About 5 to 20 ng of TAS2R + GIRK1/4 RNA was microinjected into oocytes using a microinjector. The injected oocytes were cultured in ND96 culture medium (96 mM NaCl, 2 mM KCl, 1 mM MgCl 2 , 5 mM HEPES, 1.8 mM CaCl 2 , 2.5 mM Na-pyruvate, 50 ug/mL, gentamycin, pH 7.4) for 2-3 incubated for one day (Quick & Lester, 1994). The membrane potential of oocytes was measured by measuring the peak area responding to the bitter taste of the sample using the TEVC (Two Electrode Voltage Clamp) method when high-concentration potassium was added. et al., 1995).

As can be seen in FIGS. 17a, 17b and Table 12, when frog eggs expressing bitter taste receptors were treated with single and mixed enzyme-treated dietary fiber powder at a concentration of 10 ug/mL, the result measured by the TEVC method (Current) , the untreated powder showed 10 nA, and the NY+UF+KN enzyme-treated powder showed a 50% reduction of 5 nA.

unprocessed NY UF KN NY+UF NY+UF+KN TAS2R16 Current (nA) 10 ^a ND 10 ^a 9.5 ^a 10.5 ^a 5 ^b

Example 7: Investigation of water absorption, water absorption, fat absorption, cholesterol, choline, and antioxidant activity of complex citron dietary fiber

The water absorption, swelling and liposuction power of dietary fiber absorbs much more water than its own weight or attaches to fat to increase the weight of the stool, leading to prevention of constipation and excreting bad fat from the body. In particular, dietary fiber adsorbs serum cholesterol and triglycerides to delay glucose absorption and prevents cardiovascular diseases (hyperlipidemia, arteriosclerosis, etc.) by excreting them.

In addition, dietary fiber stimulates the intestinal lining, causing constipation, forming a viscous gel solution, and reducing cholate by binding with mineral bile. Cholesterol produced by the liver becomes a raw material for bile acids. In general, bile acids are reabsorbed in the large intestine and have a bad effect on our body. Then, our body consumes more cholesterol to make the insufficient bile, and as a result, blood cholesterol has the effect of dropping.

The water absorption, water absorption, fat absorption, cholesterol and choline absorption of citron dietary fiber powder treated with untreated, single enzyme and mixed enzyme were investigated.

Water holding capacity (WHC) was measured as follows by the same method as Robertson JA et al. (2000). The powder was dispersed in 0.02% sodium azide aqueous solution for 18 hours and then centrifuged at 3,000 x g for 20 minutes (Supra 22K, Hanil Science Industrial, Incheon, Korea) to measure the precipitated weight and calculated according to the formula below.

Water absorption capacity (WRC, g/g) = [weight of sediment after centrifugation (g) - weight of dry powder (g)]/weight of dry powder (g)

Swelling power The lyophilized powder was placed in a measuring cylinder and the volume increased by swelling with 0.02% sodium azide (Sigma-Aldrich) aqueous solution for 18 hours.

As can be seen in FIGS. 18a and 18b , the water absorption power and swelling power of the powder treated with a single enzyme and a mixed enzyme rather than a single enzyme treatment are in the order of untreated < NY < UF < NY+UF+KN powder in the order of moisture absorption and swelling power appeared high.

Oil binding capacity (OBC) was measured in the same way as Zhu F et al. (2014). The powder was mixed with oil, dispersed at room temperature for 1 hour, and then centrifuged at 1,500 x g (Hanil Science Industrial), and the precipitate was filtered through a nylon mesh and weighed.

As can be seen in FIG. 18c , the liposuction power was higher in the NY-treated powder compared to the untreated, and the NY+UF+KN powder was the highest. There was no significant difference between the UF sample and the UF+NY sample.

Cholesterol binding capacity (CBC) was determined by Zhang et al. (2011), after mixing the egg yolk with water 1:10, 1.0 g dietary fiber and 25 ml egg yolk water were mixed, the pH was adjusted to 7.0, and the shaking was rotated at 120 rpm at 37°C for 3 hours. After centrifugation at 4000 g for 20 minutes, 0.02 ml of the reaction solution was taken and the cholesterol adsorbed content was investigated by the o-phthalaldehyde method.

As can be seen in FIG. 18d , the cholesterol adsorption power was increased by 20% compared to the untreated powder in the NY-treated powder and the mixed-treated powder at acidic pH 2, whereas the UF-treated powder decreased by 10%, but at pH 7, the cholesterol adsorption power was untreated. It increased in all samples compared to that, especially in the case of mixed enzyme treatment.

For sodium cholate binding capacity (CBC), 0.5 g powder was added to 0.2 g cholic acid, 100 mL 0.1 M Na-phospate buffer, pH 7. Shake rotation was performed at 120 rpm 37° C. for 3 hours. After centrifugation at 4000 g for 20 minutes, the supernatant was 620 nm and the standard curve was obtained using cholic acid, and the adsorbed content value was measured.

As can be seen in Figure 18e, cholic acid exhibits the effect of delaying the absorption of cholesterol in the body by adsorbing it with dietary fiber. Although the overall difference between cholic acid and adsorption power is insignificant, the adsorption power of NY+UF+KN powder was increased.

Antioxidant efficacy was measured by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging effect (radical scavenging effect). Sample solutions diluted by concentration in 100 uM DPPH ethanol solution were added to a 96-well plate, reacted at 37°C for 30 minutes, and absorbance at 515 nm was measured using VERSAmax, microplate reader (Molecular Devices, USA). measure The antioxidant efficacy was expressed as the radical scavenging ability (IC50) of the sample, which appeared when the absorbance decreased by 50%, and the data were obtained by repeating the experiment three times.

As can be seen in FIG. 18f , as a result of investigating the antioxidant activity, the mixed enzyme-treated powder showed the highest antioxidant activity.

unprocessed NY UF NY+UF NY+UF+KN Water absorption (g/g) 10.59 ^c 10.80 ^c 12.70 ^b 10.24 ^c 14.23 ^a Expansion force (mL/g) 14.57 ^e 21.69 ^c 25.13 ^b 20.68 ^d 27.87 ^a Liposuction (g/g) 7.85 ^c 11.09 ^b 7.74 ^c 7.82 ^c 12.21 ^a Cholesterol adsorption capacity (pH 7) (mg/g) 23.67 ^b 24.42 ^ab 27.04 ^ab 25.79 ^ab 28.80 ^a Cholesterol adsorption capacity (pH 2) (mg/g) 21.24 ^b 24.86 ^a 19.00 ^b 25.87 ^a 26.24 ^a Cholic acid adsorption capacity (mg/g) 87.94 ^a 76.21 ^bc 80.24 ^b 71.92 ^c 90.15 ^a Antioxidant activity (%) 31.80 ^b 31.61 ^b 34.17 ^a 30.60 ^b 35.74 ^a

Korea Institute of Biotechnology and Biotechnology KCTC13679BP 20181024

<110> Industry-University Cooperation Foundation Chonnam University <120> Leuconostoc mesenteroides subsp dextranicum NY203 strain, bitter taste-reduced dietary fiber prepared by using the same and functional food composition comprising the same <130> PN190402 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> 785 F primer <400> 1 ggattagata ccctggta 18 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 907 R primer <400> 2 ccgtcaattc mtttragttt 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 27 F primer <400> 3 agagtttgat cmtggctcag 20 <210> 4 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> 1492 R primer <400> 4 tacggytacc ttgttacgac tt 22

Claims (16)

Leuconostoc mesenteroids subsp. dextranicum deposited as accession number KCTC 13679BP; culture medium of the NY203 strain; Aspergillus Niger ( Aspergillus niger ) A first enzyme solution containing a strain-derived pectinase (Pectinase), glucanase (Glucanase) and arabinase (Arabinase); And Aspergillus niger strain-derived cellulase (Cellulase), hemicellulase (Hemicellulase) and a method for producing dietary fiber comprising a culturing step of culturing a second enzyme solution containing protease (protease) in the presence of vegetable fiber. The method according to claim 1, wherein the concentration of the leukonostok mesenteroid subspecies Dextranicum NY203 strain culture is 0.1 to 20.0% (v/v). delete The method of claim 1, wherein the concentration of the first enzyme solution is 0.01 to 2.00% (v/v). The method of claim 1, wherein the concentration of the second enzyme solution is 0.01 to 2.00% (v/v). The method according to claim 1, wherein the vegetable fiber is derived from the fruit of a plant. The method according to claim 1, wherein the concentration of vegetable fiber is 1 to 20% (w/v). The method according to claim 1, wherein the culturing step is performed for 1 to 15 hours. Leuconostoc mesenteroides subsp. dextranicum NY203 strain deposited with accession number KCTC 13679BP; Aspergillus Niger (Aspergillus niger) strain-derived pectinase (Pectinase), glucanase (Glucanase) and a first enzyme solution containing arabinase (Arabinase); And Aspergillus niger strain-derived cellulase (Cellulase), hemicellulase (Hemicellulase) and a second enzyme solution containing a protease (protease) in the presence of vegetable fiber by culturing a dietary fiber prepared for diet health function food composition. The method of claim 9, wherein the concentration of the leukonostok mesenteroid subspecies dextranicum NY203 strain culture is 0.1 to 20.0% (v / v), the dietary health functional food composition. delete [Claim 10] The dietary supplement composition for diet according to claim 9, wherein the vegetable fiber is derived from the fruit of a plant. Leuconostoc mesenteroides subsp. dextranicum NY203 strain deposited with accession number KCTC 13679BP; Aspergillus Niger (Aspergillus niger) strain-derived pectinase (Pectinase), glucanase (Glucanase) and a first enzyme solution containing arabinase (Arabinase); And Aspergillus niger strain-derived cellulase (Cellulase), hemicellulase (Hemicellulase) and a second enzyme solution containing protease (protease) containing dietary fiber prepared by culturing in the presence of vegetable fiber Health function for antioxidant food composition. According to claim 13, Leukono stock mesenteroid subspecies Dextranicum NY203 concentration of the strain culture medium is 0.1 to 20.0% (v / v) will, the health functional food composition for antioxidants. delete The health functional food composition for antioxidant according to claim 13, wherein the vegetable fiber is derived from the fruit of a plant.

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