Puffed grain-fermented enzyme having superoxide dismutase activity and catalase activity, production method therefor, and food composition comprising same

Abstract

The present invention relates to a puffed grain-fermented enzyme having excellent superoxide dismutase activity and catalase activity by fermenting a puffed grain by a Bacillus amyloliquefaciens strain, a production method therefor, and a food composition comprising same. According to the present invention, no food additive enzyme agent and coating agent are added, and thus it is possible to provide a puffed grain-fermented enzyme with excellent taste and economic efficiency and having superoxide dismutase activity and catalase activity.

Description

Puffed grain fermentation enzyme having superoxide dismutase activity and catalase activity, method for producing the same, and food composition containing the same

[0001] The present invention relates to a puffed grain fermentation enzyme having superoxide dismutase (SOD) activity and catalase activity, a method for producing the same, and a food composition comprising the same. More specifically, the present invention relates to a puffed grain fermentation enzyme having excellent superoxide dismutase activity and catalase activity, a method for producing the same, and a food composition comprising the same, by fermenting puffed grain with a Bacillus amyloliquefaciens strain.

[0002] One of the most important elements in our body is enzymes, which are necessary for our body to function properly, and if enzymes are absent in the body, many bodily functions such as breathing, digestion, and nutrient synthesis can be lost.

[0003] When we're young, our bodies are able to synthesize and supply abundant enzymes within themselves. However, modern lifestyle habits like smoking, drinking, high-fat, and sugar consumption stress cells, leading to cell death. This reduces enzyme synthesis, forcing the remaining cells to produce more digestive enzymes, ultimately leading to a vicious cycle of diminished endurance.

[0004] Therefore, by taking enzyme supplements to improve digestion, you can help break down the food you eat and reduce enzyme synthesis in the body, which can not only help maintain stomach health but also improve your biological rhythm.

[0005] "Enzyme food" refers to a product made by culturing edible microorganisms on plant-based raw materials to contain a large amount of enzymes, extracting enzyme-containing portions from food, or processing such foods using these as the main raw material.

[0006] These enzyme foods are known to contain various trace elements and physiologically active substances that help digestion and absorption through the production of various physiologically active substances and nutrients through the fermentation and maturation process of the enzymes and microorganisms in the food itself, and the proliferation of beneficial bacteria.

[0007] Grain fermented enzyme is a general term for enzyme foods made by culturing edible microorganisms in grain raw materials to contain a large amount of enzymes. Prior art techniques are disclosed, such as using any one grain among brown rice, soybeans, barley, and mixed grains (wheat, corn, and coix seed) as the grain raw material (Korean Patent Publication No. 10-1855125), or using rice and other rice grains, barley, wheat, rye, and oats, or miscellaneous grains such as millet, corn, foxtail millet, barley, buckwheat, and coix seed (Korean Patent Publication No. 10-2088758).

[0008] However, as in the present invention, there is no known puffed grain fermentation enzyme having excellent superoxide dismutase activity and catalase activity by fermenting puffed grain with Bacillus amyloliquefaciens strain.

[0009] (Prior art literature)

[0010] (Patent Document)

[0011] (Patent Document 0001) Republic of Korea Patent Publication No. 10-1855125

[0012] (Patent Document 0002) Republic of Korea Patent Publication No. 10-2088758

[0013] The purpose of the present invention is to provide a puffed grain fermentation enzyme having superoxide dismutase activity and catalase activity by fermenting puffed grain with a Bacillus strain.

[0014] Another object of the present invention is to provide a method for producing a puffed grain fermentation enzyme having superoxide dismutase activity and catalase activity by fermenting puffed grain with a Bacillus strain.

[0015] Another object of the present invention is to provide a food composition comprising a puffed grain fermentation enzyme having superoxide dismutase activity and catalase activity by fermenting puffed grain with a Bacillus strain.

[0016] In order to achieve the above-mentioned purpose, the present invention provides a puffed grain fermentation enzyme fermented by a Bacillus amyloliquefaciens strain and having superoxide dismutase activity and catalase activity.

[0017] As one embodiment of the present invention, the grain may be at least one selected from the group consisting of defatted soybeans, oats, brown rice, white rice, barley, sorghum, flaxseed, rice germ, adzuki beans, adzuki bean hulls, wheat, rye, and millet.

[0018] As one embodiment of the present invention, the strain may be Bacillus amyloliquefaciens NPKE6 deposited under accession number KCCM13257P.

[0019] As one embodiment of the present invention, the puffed grain fermentation enzyme has a superoxide dismutase activity inhibition rate of 37 to 88% at a concentration of 10 to 100 mg / mL, and a catalase activity of 1439 to 3276 mU / mL at a concentration of 5 to 100 mg / mL, wherein 1 Unit of the catalase activity may be an amount that decomposes 1 μmole of H2O2 at pH 7.0 and 25°C for 1 minute.

[0020] As one embodiment of the present invention, the grain may be at least one selected from the group consisting of defatted soybeans, oats, brown rice, white rice, barley, sorghum, flaxseed, rice germ, adzuki beans, adzuki bean hulls, wheat, rye, and millet.

[0021] In order to achieve the above purpose, the present invention provides a method for producing a puffed grain fermentation enzyme, comprising the steps of (a) grinding puffed grain; (b) soaking the puffed grain in water; (c) inoculating the grain with a Bacillus amyloliquefaciens strain and then mixing the same; and (d) fermenting the grain inoculated with the strain.

[0022] As one embodiment of the present invention, the manufacturing method may further include, before step (a), a step of putting grains into an extrusion expansion machine and expanding them under conditions of 80 to 120 bar at a temperature of 140 to 160°C to obtain expanded grains, and may further include, after step (d), a step of hot air drying and powdering.

[0023] As one embodiment of the present invention, the grain may be at least one selected from the group consisting of defatted soybeans, oats, brown rice, white rice, barley, sorghum, flaxseed, rice germ, adzuki beans, adzuki bean hulls, wheat, rye, and millet.

[0024] As one embodiment of the present invention, the strain may be Bacillus amyloliquefaciens NPKE6 deposited under accession number KCCM13257P.

[0025] In order to achieve the above purpose, the present invention provides a food composition comprising the puffed grain fermentation enzyme.

[0026] As one embodiment of the present invention, the food composition may be an enzyme food.

[0027] According to the present invention, since no food additive enzyme or coating agent is added, there is an advantage in that a puffed grain fermentation enzyme having excellent taste and economy and superoxide dismutase activity and catalase activity can be provided.

[0028] In addition, according to the present invention, there is an advantage in that it is possible to provide a health-oriented superfood by adding only the puffed grain fermentation enzyme and plant-based raw materials, thereby complying with the recommendations of the Ministry of Food and Drug Safety, and using grain raw materials.

[0029] Figure 1 shows the 16S rRNA base sequence of the Bacillus amyloliquefaciens NPKE6 strain (KCCM13257P) of the present invention.

[0030] Figure 2 shows a manufacturing process diagram of a puffed grain fermentation enzyme according to one embodiment of the present invention.

[0031] Figure 3 shows the DPPH radical scavenging ability of a puffed grain fermentation enzyme according to one embodiment of the present invention.

[0032] Figure 4 shows the SOD activity of a puffed grain fermentation enzyme according to one embodiment of the present invention.

[0033] Figure 5 shows the catalase activity of a puffed grain fermentation enzyme according to one embodiment of the present invention.

[0034] A first embodiment of the present invention relates to a puffed grain fermentation enzyme fermented by a Bacillus amyloliquefaciens strain and having superoxide dismutase activity and catalase activity.

[0035] The strain may be Bacillus amyloliquefaciens NPKE6 deposited under accession number KCCM13257P, and the strain may have superoxide dismutase (SOD) activity and catalase activity.

[0036] The above Bacillus amyloliquefaciens NPKE6 strain may additionally have, but is not limited to, DPPH radical scavenging activity.

[0037] In addition, the puffed grain fermentation enzyme of the present invention has a superoxide dismutase activity inhibition rate of 37 to 88% at a concentration of 10 to 100 mg / mL of the puffed grain fermentation enzyme, and a catalase activity of 1439 to 3276 mU / mL at a concentration of 5 to 100 mg / mL of the puffed grain fermentation enzyme, wherein 1 Unit of the catalase activity may be an amount that decomposes 1 μmole of H2O2 at pH 7.0 and 25°C for 1 minute.

[0038] In one embodiment of the present invention, the grain may be at least one selected from the group consisting of defatted soybeans, oats, brown rice, white rice, barley, sorghum, flaxseed, rice germ, adzuki beans, adzuki bean hulls, wheat, rye, and millet, but is not limited thereto.

[0039] A second embodiment of the present invention relates to a method for producing a puffed grain fermentation enzyme, comprising the steps of (a) grinding puffed grain; (b) soaking the puffed grain in water; (c) inoculating the grain with a Bacillus amyloliquefaciens strain and then mixing the grain; and (d) fermenting the grain inoculated with the strain.

[0040] The method for producing the puffed grain fermentation enzyme of the present invention may further include, before the step (a), a step of putting grain into an extrusion puffer and puffing it at a temperature of 140 to 160°C, preferably 145 to 155°C, more preferably 150°C, and under conditions of 80 to 120 bar, preferably 90 to 110 bar, more preferably 100 bar to obtain puffed grain.

[0041] In addition, a step of hot air drying and powdering may be further included after the above step (d), but the powdering may be formulated into a different formulation as needed.

[0042] In the method for producing the puffed grain fermentation enzyme of the present invention, the grain may be at least one selected from the group consisting of defatted soybeans, oats, brown rice, white rice, barley, sorghum, flaxseed, rice germ, adzuki beans, adzuki bean hulls, wheat, rye, and millet, but is not limited thereto.

[0043] In the method for producing the puffed grain fermentation enzyme of the present invention, the strain may be Bacillus amyloliquefaciens NPKE6 deposited under accession number KCCM13257P, but is not limited thereto.

[0044] A third embodiment of the present invention relates to a food composition comprising the above-described puffed grain fermentation enzyme.

[0045] The food composition may preferably be an enzyme food, but is not limited thereto. In addition to the puffed grain fermentation enzyme, the food composition may further include, but is not limited to, two or more types of fermented mixed grains selected from the group consisting of brown rice, white rice, sorghum, and barley; two or more types of grain mixed powders selected from the group consisting of roasted brown rice, roasted soybeans, and papaya extract; and potato extract powder.

[0046] The above food composition may include, but is not limited to, 20 to 40 wt% of the fermented grain, 50 to 70 wt% of the fermented mixed grain, 1 to 5 wt% of the mixed grain powder, and 1 to 5 wt% of the potato extract powder relative to the total weight.

[0047] The food composition may preferably include, but is not limited to, 30 to 35 wt% of the puffed grain fermentation enzyme, 55 to 65 wt% of the fermented mixed grain, 2 to 4 wt% of the grain mixed powder, and 2 to 4 wt% of the potato extract powder relative to the total weight.

[0048] The food composition may more preferably include 33 to 34 wt% of the puffed grain fermentation enzyme, 60 to 62 wt% of the fermented mixed grain, 2.5 to 3.5 wt% of the grain mixed powder, and 2.5 to 3.5 wt% of the potato extract powder relative to the total weight, but is not limited thereto.

[0049] Brown rice, which can be included in the above fermented mixed grain, is rice that has been harvested, dried, and threshed, and then the husk is removed using a machine with a rubber roller. It is known to be effective in preventing adult diseases.

[0050] The white soybeans that can be included in the above fermented mixed grains are also called meju soybeans. Nutritionally, they are particularly rich in lecithin, saponins, isoflavones, and trypsin inhibitors. These ingredients possess anticancer properties, lower blood cholesterol, inhibit fat synthesis, and thus prevent obesity. They also stimulate intestinal movement through their intestinal cleansing properties, and facilitate bowel movements, thus preventing constipation.

[0051] Sorghum, which can be included in the above fermented mixed grains, is low in fat and calories, making it an easy diet food, and has the effect of improving cholesterol levels and breaking down waste in the intestines. In addition, the proanthocyanidin contained in sorghum strengthens the immune function of the bladder and plays a role in alleviating inflammation by reducing oxidative stress on cells.

[0052] Barley, which can be included in the above fermented mixed grains, is rich in dietary fiber and helps with constipation, and its rich content of polyphenol compounds helps with antioxidant effects and improving immunity. It also contains tocotrienol, which helps lower cholesterol levels and prevents vascular diseases.

[0053] The papaya extract that can be included in the above grain mixture powder contains papain, a type of protein-decomposing enzyme extracted from the papaya fruit. The papain is one of the powerful digestive enzymes found in nature and is known to have various effects such as helping to activate digestive function, suppressing stomach bloating, promoting diuresis, and having anti-inflammation.

[0054] In the potato extract powder included in the above food composition, the potato is an alkaline food, which protects the stomach by protecting and improving ulcers and mucous membranes caused by excessive gastric acid, and contains a large amount of vitamin C (similar to spinach or tangerines), and the vitamin C of potatoes is trapped in starch, so it has a relatively high resistance to heat, and contains a high content of potassium of 410 to 485 mg per 100 g compared to other vegetables or foods, and the starch of potatoes contains a large amount of pectin, a type of dietary fiber, which has the effect of promoting intestinal function and smooth bowel movements.

[0055] Hereinafter, the present invention will be described in detail with examples and other embodiments to aid understanding. However, the embodiments according to the present invention may be modified in various different forms, and the scope of the present invention should not be construed as being limited to the following examples. The embodiments of the present invention are provided to more fully explain the present invention to those of average skill in the art.

[0056] <Example 1> Identification of Bacillus amyloliquefaciens NPKE6 strain

[0057] 1. Base sequence analysis of Bacillus amyloliquefaciens NPKE6 strain

[0058] DNA extraction and 16S rRNA sequence analysis were performed by Macrogen. Bacterial identification was performed using the Basic Local Alignment Search Tool (BLAST) search engine of the National Center for Biotechnology Information (NCBI, www.ncbi.nlm.nih.gov) using 16S rRNA sequences.

[0059] As a result of analyzing the 16S rRNA base sequence for identification and classification of the isolated strain, the base sequence of sequence number 1 was obtained.

[0060]

[0061] 2. Biochemical characteristics of Bacillus amyloliquefaciens NPKE6 strain

[0062] The biochemical characteristics of Bacillus amyloliquefaciens NPKE6 strain were investigated using API 50CH (bioMerieux Co., France), which is used for Bacillus identification. The API kit was used according to the manufacturer's instructions, and the results are shown in Table 1.

[0063] Bacillus amyloliquefaciens NPKE6 strain was inoculated into TSB (casein 17.0 g, soybean meal 3.0 g, NaCl 5.0 g, dipotassium phosphate 2.5 g, dextrose 2.5 g, final pH 7.3±0.2 at 25°C) liquid medium and cultured at 37°C. The culture was dispensed into Eppendorf tubes and centrifuged at 10,000 rpm for 5 min. The cells were harvested and washed once with sterile saline (0.85%). The cells were suspended in sterile saline and inoculated into ampoules of aseptically broken API 50CH medium, mixed evenly by pipetting, and then dispensed 150 μl into the microtubes of each test strip.

[0064] After dispensing, the API kit was cultured in a 37℃ incubator for 24 hours, and the results of color change were observed. In Table 1 below, each enzyme is indicated with a + if used and a - if not used.

[0065]

[0066] As a result of identification, based on the above biochemical characteristics, the selected NPKE6 strain was identified as Bacillus amyloliquefaciens and named Bacillus amyloliquefaciens NPKE6, and was deposited at the Korean Culture Center of Microorganisms (KCCM) on October 31, 2022, and assigned the accession number KCCM13257P.

[0067]

[0068] <Example 2> Production of grain fermentation enzyme using Bacillus amyloliquefaciens NPKE6 strain

[0069] 1. Cultivation of strains

[0070] Bacillus amyloliquefaciens NPKE6 was inoculated (1.0% v / v) into BM (D-Glucose monohydrate (13.5 g / L), Hydrolysed soy protein (3 g / L), Yeast extract (3 g / L), Sodium carbonate (0.3 g / L), Magnesium sulfate heptahydrate (0.2 g / L)) medium and cultured at 30°C for 24 hours.

[0071]

[0072] 2. Production of puffed grains

[0073] After weighing the grains, 18% water was added and mixed. The grains were expanded into 2.5 cm x 2.5 cm particles at 150°C and 100 bar, and cooling water at 30°C was used.

[0074]

[0075] 3. Production of puffed grain fermentation enzyme

[0076] After weighing the puffed grains, 9 times the amount of 70℃ sterilized water was added, and after soaking for 20 minutes, the unabsorbed sterilized water was removed. 2.5 kg of the soaked material was added per tray, and the Bacillus amyloliquefaciens NPKE6 inoculum, which had been cultured in BM medium at 30℃ and 150 rpm for 24 hours, was inoculated at an inoculation rate of 2% (v / w) based on the soaked material.

[0077] After fermentation for 24 hours at 30℃ and a relative humidity of 80% or higher, the grain fermentation enzyme was manufactured by drying with hot air at 70℃ for 24 hours and grinding to a size of 60 mesh using a multipurpose grinder.

[0078]

[0079] <Example 3> Analysis of enzyme activity of puffed grain fermentation enzyme

[0080] 1. Confirmation of DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging ability

[0081] The puffed grain fermentation enzyme prepared in Example 2 was diluted with deionized water according to concentration, centrifuged at 4000 rpm for 20 minutes, and filtered through a 0.2 μm filter to prepare a puffed grain fermentation enzyme filtrate.

[0082] After mixing 500 μL of 0.2 mM DPPH (2,2-diphenyl-1-picrylhydrazyl) solution and 420 μL of ethanol with 80 μL of diluted sample by concentration, the absorbance was measured at 517 nm, and the DPPH radical scavenging ability was calculated using the following formula. The results are shown in Figure 3.

[0083] DPPH radical scavenging activity (%) = {1-(absorbance of sample addition group / absorbance of negative control group)}*100

[0084] As shown in Fig. 3, it can be confirmed that the DPPH radical scavenging activity of the puffed grain fermentation enzyme increases in a concentration-dependent manner.

[0085]

[0086] 2. SOD activity

[0087] The SOD activity of the puffed grain fermentation enzyme prepared in Example 2 was measured using the EZ-SOD Assay Kit (DoGen, Korea) according to the manufacturer's manual.

[0088] That is, the puffed grain fermentation enzyme was diluted with sterilized water according to concentration, centrifuged at 4000 rpm for 20 minutes, and filtered through a 0.2 μm filter to prepare a puffed grain fermentation enzyme filtrate. As shown in Table 2 below, 20 μL of the diluted sample according to concentration was added to each sample well and the well of Blank 2. At this time, 20 μL of ddH2O was added to each of Blank 1 and Blank 3 wells.

[0089] 200 μL of WST working solution was added to each well. 20 μL of dilution buffer was added to each of wells Blank 2 and Blank 3. Using a Malti-channel pipette, 20 μL of enzyme working solution was added to each of wells Blank 1 and sample and mixed carefully. The 96-well plate was incubated at 37°C for 20 minutes.

[0090] Blank 1Blank 2Blank 3Test sampleSample-20 μL-20 μLddH2O20 μL-20 μL-WST working solution200 μL200 μL200 μL200 μLDilution buffer-20 μL20 μL-Enzyme working solution20 μL-20 μLTotal volume240 μL240 μL240 μL240 μL

[0091] After measuring the absorbance at 450 nm, the SOD activity of the puffed grain fermentation enzyme calculated using the formula below is shown in Figure 4.

[0092] SOD activity (inhibition rate %) = {(OD blank1 - OD blank3 ) - (OD sample - OD blank2 )} / (OD blank1 - OD blank3 )*100

[0093] As shown in Fig. 4, it can be confirmed that the SOD activity of the puffed grain fermentation enzyme increases in a concentration-dependent manner.

[0094]

[0095] 3. Catalase activity

[0096] The catalase activity of the puffed grain fermentation enzyme prepared in Example 2 was measured using the EZ-Catalase Assay Kit (DoGen, Korea) according to the manufacturer's manual.

[0097] That is, the puffed grain fermentation enzyme was diluted with sterilized water according to concentration, centrifuged at 4000 rpm for 20 minutes, and filtered through a 0.2 μm filter to prepare a puffed grain fermentation enzyme filtrate. 25 μL of the diluted sample according to concentration and 25 μL of a 40 μM H2O2 solution were added to a 96-well plate.

[0098] After the plate was incubated at room temperature for 30 minutes in a light-blocked environment, 50 μL of Oxi-Probe / HRP Working solution, prepared by mixing 30 μL of 10 mM Oxi-Probe, 12 μL of 100 U / mL horseradish-peroxide (HRP), and 3 mL of 1X reaction buffer, was added to each well of the plate where the reaction was completed.

[0099] After reacting for 30 minutes in a light-blocking 37℃ place, the absorbance of the plate after the reaction was measured at 560 nm. A standard calibration curve (y = 23673x - 224.98, R) was prepared using catalase measured at each concentration as a standard substance. 2 = 0.999) was substituted into the equation.

[0100] The catalase activity (unit: mU / mL) of the puffed grain fermentation enzyme is shown in Figure 5. As can be seen in Figure 5, the catalase activity of the puffed grain fermentation enzyme increases in a concentration-dependent manner.

[0101]

[0102] <Example 4> Measurement of the number of viable Bacillus amyloliquefaciens in puffed grain fermentation enzyme

[0103] The sample was analyzed using a 10-fold dilution solution using saline. The viable cell count was measured by counting the colony count after spreading 100 μL of the prepared dilution solution on sterilized and dried PCA solid medium and incubating it in a 37°C incubator for 16 hours.

[0104] The measured viable Bacillus amyloliquefaciens counts are shown in Table 3 below.

[0105] Distinctive viable cell count (CFU / g) Defatted soybean meal 3.0 x10 8 Puffed defatted soybean meal 3.7 x10 8

[0106] As described above, specific parts of the present invention have been described in detail. It will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments and that the scope of the present invention is not limited thereby.

[0107] Accordingly, the substantial scope of the present invention is defined by the appended claims and their equivalents. Simple modifications and variations of the present invention can be readily employed by those skilled in the art, and all such modifications and variations are considered to fall within the scope of the present invention.

[0108] [Accession number]

[0109] Name of depositor: Korea Center for Microbiological Conservation (KCCM)

[0110] Address of the depository: Yulim Building, 45 Hongje-nae 2-ga-gil, Seodaemun-gu, Seoul

[0111] Deposit date: 20221031

[0112] Accession number: KCCM13257P

[0113]

Claims

1. A puffed grain fermentation enzyme fermented by Bacillus amyloliquefaciens strain and having superoxide dismutase activity and catalase activity.

2. A puffed grain fermentation enzyme according to claim 1, characterized in that the strain is Bacillus amyloliquefaciens NPKE6 deposited under the accession number KCCM13257P.

3. In paragraph 1, Superoxide dismutase activity inhibition rate of 37-88% at a concentration of 10-100 mg / mL of the above-mentioned puffed grain fermentation enzyme, and The above puffed grain fermentation enzyme has a catalase activity of 1439 to 3276 mU / mL at a concentration of 5 to 100 mg / mL, 1 Unit of the above catalase activity is equivalent to 1 minute at pH 7.0, 25°C in H 2 O 2 A puffed grain fermentation enzyme characterized by an amount that decomposes 1 μmole.

4. A puffed grain fermentation enzyme in paragraph 1, wherein the grain is at least one selected from the group consisting of defatted soybeans, oats, brown rice, white rice, barley, sorghum, flaxseed, rice germ, adzuki beans, adzuki bean hulls, wheat, rye, and millet. 5.(a) Step of grinding the puffed grains; (b) a step of immersing the above-mentioned puffed grains in water; (c) a step of inoculating the grain with the Bacillus amyloliquefaciens strain and then mixing; and (d) A method for producing a puffed grain fermentation enzyme, comprising a step of fermenting grain inoculated with the above strain.

6. In the fifth paragraph, before step (a), a step of putting grain into an extrusion expansion machine and expanding it under conditions of 80 to 120 bar at a temperature of 140 to 160°C is further included to obtain expanded grain. A method for producing a puffed grain fermentation enzyme, characterized in that it further comprises a step of hot air drying and powdering after the step (d) above.

7. A method for producing a puffed grain fermentation enzyme, characterized in that in paragraph 5, the grain is at least one selected from the group consisting of defatted soybeans, oats, brown rice, white rice, barley, sorghum, flaxseed, rice germ, adzuki beans, adzuki bean hulls, wheat, rye, and millet.

8. A method for producing a puffed grain fermentation enzyme, characterized in that the strain in paragraph 5 is Bacillus amyloliquefaciens NPKE6 deposited under the accession number KCCM13257P.

9. A food composition comprising a puffed grain fermentation enzyme according to any one of claims 1 to 4.

10. A food composition according to claim 9, characterized in that the food composition is an enzyme food.

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