WO2024095969A1 - Aliment fermenté, procédé de production d'aliment fermenté, et technologie de suppression d'odeur de fermentation pour aliment fermenté - Google Patents
Aliment fermenté, procédé de production d'aliment fermenté, et technologie de suppression d'odeur de fermentation pour aliment fermenté Download PDFInfo
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- NQXWGWZJXJUMQB-UHFFFAOYSA-K iron trichloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].Cl[Fe+]Cl NQXWGWZJXJUMQB-UHFFFAOYSA-K 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 235000021332 kidney beans Nutrition 0.000 description 1
- 239000000252 konjac Substances 0.000 description 1
- 235000010485 konjac Nutrition 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 235000021056 liquid food Nutrition 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 1
- CNFDGXZLMLFIJV-UHFFFAOYSA-L manganese(II) chloride tetrahydrate Chemical compound O.O.O.O.[Cl-].[Cl-].[Mn+2] CNFDGXZLMLFIJV-UHFFFAOYSA-L 0.000 description 1
- 230000000873 masking effect Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000010460 mustard Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 235000015927 pasta Nutrition 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 235000011197 perejil Nutrition 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 235000020233 pistachio Nutrition 0.000 description 1
- 238000005498 polishing Methods 0.000 description 1
- 108700022290 poly(gamma-glutamic acid) Proteins 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 229940093916 potassium phosphate Drugs 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 235000013406 prebiotics Nutrition 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000001902 propagating effect Effects 0.000 description 1
- 235000020236 pumpkin seed Nutrition 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
- 235000021487 ready-to-eat food Nutrition 0.000 description 1
- HELXLJCILKEWJH-NCGAPWICSA-N rebaudioside A Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HELXLJCILKEWJH-NCGAPWICSA-N 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 235000002020 sage Nutrition 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- WBHQBSYUUJJSRZ-UHFFFAOYSA-M sodium bisulfate Chemical compound [Na+].OS([O-])(=O)=O WBHQBSYUUJJSRZ-UHFFFAOYSA-M 0.000 description 1
- 229910000342 sodium bisulfate Inorganic materials 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 229940001516 sodium nitrate Drugs 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000007901 soft capsule Substances 0.000 description 1
- 239000008279 sol Substances 0.000 description 1
- 235000021055 solid food Nutrition 0.000 description 1
- 235000013555 soy sauce Nutrition 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 235000020238 sunflower seed Nutrition 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 239000001585 thymus vulgaris Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000020234 walnut Nutrition 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/50—Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L25/00—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
- A23L7/104—Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D13/00—Finished or partly finished bakery products
- A21D13/80—Pastry not otherwise provided for elsewhere, e.g. cakes, biscuits or cookies
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
- A23L7/109—Types of pasta, e.g. macaroni or noodles
Definitions
- the present invention relates to fermented foods, methods for producing fermented foods, and technology for suppressing the fermentation odor of fermented foods.
- Fermented foods such as natto
- the bacteria used in fermentation and the enzymes they produce have probiotic or prebiotic properties, such as intestinal regulating effects, making fermented foods useful as ready-to-eat foods that contribute to health. For this reason, there is a demand for the development of fermented foods that can be consumed on a daily basis.
- Soybean natto has a grassy smell derived from soybeans and a fermented smell that is unique to natto (for example, Patent Document 1). These flavors are strong, making it difficult to eat natto on a daily basis.
- the objective of the present invention is to provide a fermented food with reduced fermentation odor and grassy odor.
- a fermented food contains a fermented product of at least one edible plant selected from the group consisting of nuts and seeds, grains, beans, vegetables, potatoes, mushrooms, and fruits, and has a methyl palmitate content of 0.01 ppb or more and/or an ethyl palmitate content of 0.01 ppb or more. It has been found that the above-mentioned problems can be solved. Based on this finding, the present inventors have conducted further research and have completed the present invention. That is, the present invention includes the following aspects.
- a fermented food comprising a fermentation product of at least one edible plant selected from the group consisting of beans, grains, vegetables, potatoes, nuts, seeds, mushrooms and fruits, and having a methyl palmitate content of 0.01 ppb or more, or 0.05 ppb or more, or 0.1 ppb or more, or 1.0 ppb or more, while there is no particular upper limit to the content, for example, 10,000 ppb or less, or 1,000 ppb or less, or 500 ppb or less, or 100 ppb or less, and/or an ethyl palmitate content of 0.01 ppb or more, or 0.05 ppb or more, or 0.1 ppb or more, or 0.5 ppb or more, or 1.0 ppb or more, while there is no particular upper limit to the content, for example, 10,000 ppb or less, or 1,000 ppb or less, or 500 ppb or less, or 100 ppb or less.
- Item 2 The fermented food according to Item 1, wherein the starch content of the edible plant is 4% by mass or more, or 10% by mass or more, or 15% by mass or more, or 20% by mass or more, or 25% by mass or more, or 30% by mass or more, while the upper limit of the content is, for example, 95% by mass or less, or 90% by mass or less, or 80% by mass or less, or 70% by mass or less.
- the starch content of the edible plant is 4% by mass or more, or 10% by mass or more, or 15% by mass or more, or 20% by mass or more, or 25% by mass or more, or 30% by mass or more, while the upper limit of the content is, for example, 95% by mass or less, or 90% by mass or less, or 80% by mass or less, or 70% by mass or less.
- the fermented food according to any one of the above items relating to a fermented food, wherein the palmitic acid content of the edible plant is 1 ppm or more, or 10 ppm or more, or 100 ppm or more, or 1000 ppm or more, or 1500 ppm or more, or 2000 ppm or more, or 2500 ppm or more, while the upper limit of the content is not particularly limited, for example, 10,000,000 ppm or less, or 1,000,000 ppm or less, or 100,000 ppm or less.
- the fermented food according to any one of the above items, which has an ⁇ -amylase activity of 2 U/g or more, or 3 U/g or more, or 4 U/g or more, or 5 U/g or more, or 6 U/g or more, or 7 U/g or more, or 8 U/g or more, or 9 U/g or more, or 10 U/g or more, or 15 U/g or more, or 20 U/g or more, or 30 U/g or more, or 40 U/g or more, or 50 U/g or more, or 60 U/g or more, or 70 U/g or more, or 80 U/g or more, or 90 U/g or more, or 100 U/g or more, or 150 U/g or more, or 200 U/g or more, and the upper limit is not particularly limited, but is usually 100,000 U/g or less, or 80,000 U/g or less.
- Item 5 The fermented food according to any one of the above items relating to a fermented food, wherein the mass ratio of the ethyl palmitate to the methyl palmitate (ethyl palmitate mass/methyl palmitate mass) is 0.0001 to 10,000, the lower limit of the ratio is 0.0001 or more, or 0.001 or more, or 0.01 or more, or 0.1 or more, and the upper limit is not particularly limited, but is usually 10,000 or less, or 1,000 or less, or 100 or less, or 10 or less.
- the mass ratio of the ethyl palmitate to the methyl palmitate is 0.0001 to 10,000
- the lower limit of the ratio is 0.0001 or more, or 0.001 or more, or 0.01 or more, or 0.1 or more
- the upper limit is not particularly limited, but is usually 10,000 or less, or 1,000 or less, or 100 or less, or 10 or less.
- the legume is at least one kind selected from the group consisting of the genus Pisum, the genus Phaseolus, the genus Pigeonpea, the genus Vitis, the genus Vicia, the genus Chickpea, the genus Glycine, and the genus Lens.
- Item 7 The fermented food according to any one of the above items relating to fermented foods, wherein the cereal is at least one selected from the group consisting of the genus Oryza, the genus Maize, the genus Barley, and the genus Triticum.
- Item 9 The fermented food according to Item 8, wherein the millet is at least one selected from the group consisting of foxtail millet, barnyard millet, millet, sorghum, rye, oats, pearl barley, corn, buckwheat, amaranth, and quinoa.
- Item 10 The fermented food according to any one of the above items relating to fermented foods, wherein the vegetable is at least one type selected from the group consisting of the Solanum genus and the Cucurbita genus.
- the fermented food according to any one of the above items relating to a fermented food, wherein the mass ratio of soluble carbohydrates to starch (soluble carbohydrate mass/starch mass) is 0.05 or more, or 0.1 or more, or 0.2 or more, or 0.4 or more, or 0.6 or more, or 0.8 or more, and the upper limit of the mass ratio is 10 or less, or 8 or less, or 6 or less.
- the mass ratio of soluble carbohydrates to starch is 0.05 or more, or 0.1 or more, or 0.2 or more, or 0.4 or more, or 0.6 or more, or 0.8 or more, and the upper limit of the mass ratio is 10 or less, or 8 or less, or 6 or less.
- the fermented food according to any one of the above items which is related to a fermented food, has a soluble carbohydrate content of 2.0% by mass or more, or 4.0% by mass or more, or 5.0% by mass or more, or 7.0% by mass or more, or 8.0% by mass or more, or 10% by mass or more, and the upper limit of the content is not particularly limited, but is 50% by mass or less, or 40% by mass or less, or 30% by mass or less, or 25% by mass or less.
- Item 13 The fermented food according to any one of the above items, which contains a bacterium capable of producing PGA. Item 14.
- the viable cell count of the bacteria capable of producing PGA is 1.0 x 10 cells/g or more, or 2.0 x 10 cells/g or more, or 3.0 x 10 cells/g or more, or 4.0 x 10 cells/g or more, and the upper limit is 1.0 x 10 cells/g or less, or 8.0 x 10 cells/g or less, or 6.0 x 10 cells/g or less, or 4.0 x 10 cells/g or less.
- N number ⁇ ⁇ number ⁇ Value ⁇ : PGA content (mg) per 1 g of fermented food Value ⁇ : Levan content (mg) per 1 g of fermented food Item 16.
- the fermented food according to any one of the above items relating to a fermented food, wherein the viscosity of an extract obtained by stirring 40 g of the fermented food in 40 mL of water and then removing solids is 180 mPa ⁇ s or less, 170 mPa ⁇ s or less, or 160 mPa ⁇ s or less under conditions of a B-type viscometer (60 rpm, 25° C., rotor No. 2). Item 17.
- a fermented food according to any of the above items relating to a fermented food in which when two grains of the fermented food are pressed tightly together and then one grain is lifted upward at a speed of 100 mm/min, the distance until a thread formed between the two grains of the fermented food breaks is 100 mm or less, or 50 mm or less, or 25 mm or less, or 15 mm or less, or 10 mm or less, or 5 mm or less.
- Item 18 The fermented food according to any one of Items 15 to 17, wherein the fermented food is an animal feed.
- a method for producing a fermented food according to any one of the above items relating to a fermented food comprising a step of fermenting a composition containing at least one edible plant selected from the group consisting of nuts and seeds, grains, beans, vegetables, potatoes, mushrooms, and fruits, and koji, and having a salt content of 1000 mg or less, or 500 mg or less, or 300 mg or less, or 100 mg or less, or 50 mg or less per 100 g, with the lower limit not being particularly limited, for example, 1 mg or more, or 2 mg or more, or 3 mg or more, at a product temperature of 30° C. to 60° C.
- Item 20 The method according to item 19, wherein the composition has a dry weight moisture content of 20% by mass or more, or 30% by mass or more, or 40% by mass or more, or 50% by mass or more, and the upper limit of the dry weight moisture content is not limited, but is, for example, 90% by mass or less, 80% by mass or less, or 70% by mass or less.
- Item 21 The method according to any one of the above items, wherein the edible plant is an edible plant that has been heat-treated at 80° C. or higher under conditions of a moisture content of 50% or more by mass on a dry basis.
- Item 22 The method according to any one of the above items, wherein the starch content of the composition is 4% by mass or more, or 10% by mass or more, or 15% by mass or more, or 20% by mass or more, or 25% by mass or more, or 30% by mass or more, and the upper limit of the starch content is 95% by mass or less, or 90% by mass or less, or 80% by mass or less, or 70% by mass or less, or 60% by mass or less, or 50% by mass or less, or 40% by mass or less.
- the starch content of the composition is 4% by mass or more, or 10% by mass or more, or 15% by mass or more, or 20% by mass or more, or 25% by mass or more, or 30% by mass or more, and the upper limit of the starch content is 95% by mass or less, or 90% by mass or less, or 80% by mass or less, or 70% by mass or less, or 60% by mass or less, or 50% by mass or less, or 40% by mass or less.
- the ⁇ -amylase activity of the composition is 2 U/g or more, or 10 U/g or more, or 15 U/g or more, and although there is no particular upper limit, it is usually 20,000 U/g or less, or 5,000 U/g or less, or 1,000 U/g or less, or 500 U/g or less.
- Item 24 The method according to any one of the above items, wherein the mass ratio of the koji to the edible plant (mass of koji/mass of edible plant) is 0.1 or more, or 0.15 or more, with the upper limit being 0.3 or less, or 0.25 or less.
- the moisture content of the koji on a dry basis is 50% by mass or less, or 40% by mass or less, or 30% by mass or less, or 20% by mass or less, or 15% by mass or less, or 10% by mass or less, or 5% by mass or less, and the lower limit is not particularly limited, but is, for example, 0.1% by mass or more, or 0.5% by mass or more, or 1% by mass or more.
- the lower limit is not particularly limited, but is, for example, 0.1% by mass or more, or 0.5% by mass or more, or 1% by mass or more.
- the method according to any one of the above items wherein the d50 of the koji after ultrasonic treatment is 1000 ⁇ m or less, or 750 ⁇ m or less, or 500 ⁇ m or less, or 400 ⁇ m or less, or 350 ⁇ m or less, and although the lower limit is not particularly specified, is 0.1 ⁇ m or more, or 1 ⁇ m or more.
- Item 27. The method according to any one of the above items, which relates to a production method comprising a step of fermenting with a bacterium capable of producing PGA. Item 28. In the state where the number of the bacteria in the step (II) is 1.0 x 10 1 to 3.0 x 10 3 /g, the average fermentation temperature is more than 40 ° C.
- Item 28 The method for producing the same according to item 27.
- Item 29A The method according to any one of the above items, wherein the bacterium capable of producing PGA in the step (II) is a bacterium belonging to the genus Bacillus.
- Item 29B The method according to Item 29A, wherein the Bacillus bacterium is Bacillus subtilis natto bacteria or Bacillus velezensis bacillus subtilis.
- Item 29C The method according to Item 29B, wherein the natto bacteria belonging to Bacillus subtilis are Bacillus subtilis var. natto, Bacillus subtilis (natto), or Bacillus natto.
- Item 30 The method according to any one of the above items, wherein the bacterium capable of producing PGA in the step (II) is Bacillus subtilis natto. Item 31.
- the method comprises fermenting a composition containing beans, koji, and Bacillus subtilis natto, which have been heat-treated at 80 ° C. or higher under conditions of a moisture content of 50% by mass or higher on a dry basis,
- the average fermentation temperature is more than 40°C and less than 65°C and the average soluble carbohydrate content is less than 2.0 mass%
- the average fermentation temperature is more than 45°C and less than 75°C and the average soluble carbohydrate content is 2.0 mass% or more.
- Item 32 The fermented food according to any one of the above items relating to a fermented food, which is in a paste or liquid form.
- Item 33 A processed grain product comprising the paste-like or liquid fermented food according to Item 32.
- Item 34 The processed grain product according to Item 33, which is selected from the group consisting of cooked rice, noodles, bread, waffles, cereals, and beverages.
- Item 35 An agent for suppressing the fermented odor and grassy odor of fermented foods, comprising at least one selected from the group consisting of methyl palmitate and ethyl palmitate.
- Item 36 An agent for suppressing the fermented odor and grassy odor of fermented foods, comprising at least one selected from the group consisting of methyl palmitate and ethyl palmitate.
- a method for producing a fermented food having a suppressed fermentation odor and grassy odor comprising adjusting the content of methyl palmitate in the fermented food to 0.01 ppb or more, or 0.05 ppb or more, or 0.1 ppb or more, or 1.0 ppb or more, while there is no particular upper limit to the content, for example, 10,000 ppb or less, or 1,000 ppb or less, or 500 ppb or less, or 100 ppb or less, and/or adjusting the content of ethyl palmitate to 0.01 ppb or more, or 0.05 ppb or more, or 0.1 ppb or more, or 0.5 ppb or more, or 1.0 ppb or more, while there is no particular upper limit to the content, for example, 10,000 ppb or less, or 1,000 ppb or less, or 500 ppb or less, or 100 ppb or less.
- the present invention can provide a fermented food in which the fermented odor and grassy odor are further suppressed.
- a fermented food in which stringiness is further suppressed can be provided.
- ppb and ppm refer to mass concentration (w/w) and represent the mass of the target component relative to the total mass of the food. Additionally, in this specification, “mass %” (sometimes written as w/w %) represents the mass of the target component relative to the total mass of the food, expressed as a percentage.
- water content on a dry basis refers to the ratio of the total amount of water derived from the raw materials of the fermented food of the present invention and the amount of water added separately to the total amount of solids.
- the value is measured by heating to 90°C using a reduced pressure heat drying method in accordance with the 2015 edition (7th edition) of the Standard Tables of Food Composition in Japan. For example, it can be measured by the following method.
- the present invention relates to a fermented food (sometimes referred to as the "fermented food of the present invention” in this specification) that contains a fermented product of at least one edible plant selected from the group consisting of nuts and seeds, cereals, beans, vegetables, potatoes, mushrooms, and fruits, and has a methyl palmitate content of 0.01 ppb or more and/or an ethyl palmitate content of 0.01 ppb or more.
- “fermentation” refers to a process in which the components in a food are changed by endogenous enzymes of a microorganism to produce an organic substance, and includes a fermentation process accompanying the growth of a microorganism and a fermentation process by endogenous enzymes possessed by a microorganism.
- “fermented products” include metabolic products produced by the growth of a microorganism and reaction products produced by endogenous enzymes possessed by a microorganism.
- the enzymes When fermentation is performed by endogenous enzymes possessed by a microorganism, the enzymes may be in any state, but may be those in which the endogenous enzymes possessed by the microorganism are isolated and produced, or in the state of inactivated bacteria (typically dead bacteria) in which the proliferation activity of the microorganism has been lost but the endogenous enzymes maintain their activity. This will be explained below.
- the fermented food of the present invention contains methyl palmitate (CAS registration number: 112-39-0) and/or ethyl palmitate (CAS registration number: 628-97-7). Although the principle is unclear, it is thought that the fermentation odor and grassy odor are suppressed by the methyl palmitate and/or ethyl palmitate masking the fermentation odor and grassy odor.
- the fermented food of the present invention only needs to contain a predetermined content of methyl palmitate and/or ethyl palmitate, it is more preferable that the fermented food of the present invention contains at least methyl palmitate at a predetermined content, and it is most preferable that the fermented food of the present invention contains both methyl palmitate and ethyl palmitate at a predetermined content. Furthermore, when the two components, methyl palmitate and ethyl palmitate, are contained, it is preferable to set the mass ratio of ethyl palmitate to methyl palmitate (ethyl palmitate mass/methyl palmitate mass) to 0.0001 to 10,000, since this will synergistically produce the effects of the present invention.
- the lower limit of the ratio is 0.0001 or more, preferably 0.001 or more, more preferably 0.01 or more, and even more preferably 0.1 or more.
- the upper limit is not particularly limited, but is usually 10,000 or less, preferably 1,000 or less, more preferably 100 or less, and even more preferably 10 or less.
- the contents of methyl palmitate and ethyl palmitate in the fermented food of the present invention can be quantified by the dynamic headspace (DHS) injection method using a gas chromatography mass spectrometer (GC/MS) as follows.
- a standard sample of each component with known content is diluted with 99.5% ethanol to 1000 ppm, and then further diluted with ultrapure water to an appropriate content (diluted standard) and the sample are analyzed. 1 g of the diluted standard and sample are placed in a 10 ml vial for analysis.
- the components in the sample can be quantified by comparing the integration results of the peak areas of the most abundant ions in the diluted standard and sample at the retention time considered to be the target component when compared with the retention time of the standard, using analysis based on the mass spectrum pattern of a mass spectrometer.
- m/z refers to the value detected in the range of -0.3 to +0.7 at the central m/z value of each component.
- concentration of each component contained in each sample is calculated from the peak area of each component obtained, taking into account the dilution rate with the solvent.
- the fermented food of the present invention may have a methyl palmitate content within a specified range.
- the methyl palmitate may be contained in food materials such as edible plants that are the raw materials for the fermented food of the present invention, may be added separately from the food materials, may be generated during the production of the fermented food of the present invention, or may be a combination of these.
- a liquid seasoning containing methyl palmitate may be added to a fermented bean food such as natto.
- the content may be, for example, 0.01 ppb to 10,000 ppb.
- the lower limit of the content is, for example, 0.01 ppb or more, preferably 0.05 ppb or more, more preferably 0.1 ppb or more, and even more preferably 1.0 ppb or more.
- the upper limit of the content is preferably 1,000 ppb or less, more preferably 500 ppb or less, and particularly preferably 100 ppb or less.
- the fermented food of the present invention may have an ethyl palmitate content within a specified range.
- the ethyl palmitate may be contained in food materials such as edible plants that are the raw materials for the fermented food of the present invention, may be added separately from the food materials, may be generated during the production of the fermented food of the present invention, or may be a combination of these.
- a liquid seasoning containing ethyl palmitate may be added to a fermented bean food such as natto.
- the content may be, for example, 0.01 ppb to 10,000 ppb.
- the lower limit of the content is, for example, 0.01 ppb or more, preferably 0.05 ppb or more, more preferably 0.1 ppb or more, even more preferably 0.5 ppb or more, and even more preferably 1.0 ppb or more.
- the upper limit of the content is preferably 1,000 ppb or less, more preferably 500 ppb or less, and particularly preferably 100 ppb or less.
- the fermented food of the present invention contains edible plants as the raw ingredients.
- the edible plant contains at least one edible plant selected from the group consisting of nuts and seeds, grains, beans, vegetables, potatoes, mushrooms, and fruits. More preferably, there may be two or more, three or more, or four or more edible plants.
- the upper limit of the number of edible plant species is not particularly limited, but may be, for example, 10 or less. It is preferable that the edible plant contains beans.
- the beans are not limited thereto, but it is preferable to use one or more kinds selected from beans of the genus Pisum, Glycine, Phaseolus, Pigeonpea, Vigna, Vicia, Chickpea, Lentil, Lupin, Lathyrium, Cucumis, Mumpsville, Carob, and Parkia, etc. Among them, it is preferable to use one or more kinds selected from beans of the genus Pisum, Phaseolus, Pigeonpea, Vigna, Vicia, Chickpea, Glycine, and Lentil.
- beans include, but are not limited to, peas (especially yellow peas, white peas, and green peas, which are immature seeds), kidney beans, red beans, white beans, black beans, pinto beans, tiger beans, lima beans, scarlet beans, pigeon peas, mung beans, cowpeas, adzuki beans, broad beans, soybeans (including edamame, which are immature seeds of soybeans harvested with the pods in an immature state and characterized by the green appearance of the beans), chickpeas, lentils, lentils, blue peas, purple peas, lentils, peanuts, lupine beans, grass peas, carob, pampas grass, long-leaved pampas grass, coffee beans, cacao beans, and Mexican jack beans.
- peas especially yellow peas, white peas, and green peas, which are immature seeds
- kidney beans red beans, white beans, black beans, pinto beans,
- the classification of edible plants can be understood by those skilled in the art who handle food and processed food products. For example, a more clear determination can be made by referring to the Standard Tables of Food Composition in Japan, 2015 Edition (7th Edition). In addition, even for foodstuffs whose edible parts (such as edamame and green peas) are treated as vegetables, it can be determined whether they are legumes or not based on the state of the whole plant (such as soybeans and peas) combined with the inedible parts (such as pods).
- the beans are preferably beans with a certain amount of starch content.
- the lower limit is 10% by mass or more, 15% by mass or more, 20% by mass or more, 25% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more
- the upper limit is not particularly limited, but may be 80% by mass or less, 70% by mass or less, or 60% by mass or less.
- the genus Pisum such as pea
- the genus Chickpea such as chickpea
- the genus Phaseolus such as kidney bean.
- the content of the genus Pisum, the genus Chickpea, and the genus Phaseolus may be a predetermined percentage or more relative to the entire fermented food, and more specifically, the content may be 5% by mass or more, 10% by mass or more, 15% by mass or more, 20% by mass or more, 25% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, 90% by mass or more, or 100% by mass of the entire fermented food.
- the content of the genus Pisum, genus Chickpea, and genus Phaseolus may be a specified percentage or more relative to the total amount of beans, and more specifically may be 5% by mass or more, 10% by mass or more, 15% by mass or more, 20% by mass or more, 25% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, 90% by mass or more, or 100% by mass of the total amount of beans.
- the content of soybeans may be a specified percentage or less of the entire fermented food, and more specifically, may be 50% by mass or less, 40% by mass or less, 30% by mass or less, 20% by mass or less, 10% by mass or less, 5% by mass or less, or 1% by mass or less of the entire fermented food.
- the content of soybeans may be a specified percentage or less relative to the total amount of beans, and more specifically, it may be 80% by mass or less, 70% by mass or less, 60% by mass or less, 50% by mass or less, 40% by mass or less, 30% by mass or less, 20% by mass or more, 10% by mass or less, 5% by mass or less, 1% by mass or less, or even 0% by mass of the total amount of beans.
- the starch content derived from the genus Pisum, Chickpea, or Phaseolus may be a specified percentage or more of the starch content of the entire fermented food, and more specifically, the starch content of the entire fermented food may be 5% by mass or more, 10% by mass or more, 15% by mass or more, 20% by mass or more, 25% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, 90% by mass or more, or 100% by mass.
- the starch content derived from the genus Glycine may be a specified percentage or less relative to the total fermented food, and more specifically, the starch content of the total fermented food may be 80% by mass or less, 70% by mass or less, 60% by mass or less, 50% by mass or less, 40% by mass or less, 30% by mass or less, 20% by mass or less, 10% by mass or less, 5% by mass or less, 1% by mass or less, or even 0% by mass.
- the cereal is preferably one or more selected from the genus Oryza, genus Maize, genus Hordeum, genus Triticum, etc., but is not limited thereto.
- Specific examples of cereals include, but are not limited to, amaranth, foxtail millet, oats, barley, millet, quinoa, wheat, rice, sugarcane, buckwheat, corn, pearl barley, barley, fonio, sorghum, etc.
- the cereal is preferably a miscellaneous cereal described below, and for example, oats or corn (particularly sweet corn) is preferable.
- miscellaneous grains refers to grains other than the major grains rice, wheat, and barley among the above-mentioned grains, and is a concept including pseudo-miscellaneous grains (Chenopodiaceae, Amaranthaceae) other than the so-called Poaceae grains.
- the type of miscellaneous grains used is not limited, but is preferably at least one type of miscellaneous grain selected from the group consisting of Poaceae, Chenopodiaceae, and Amaranthaceae, and is more preferably Poaceae.
- miscellaneous grains include, but are not limited to, foxtail millet, barnyard millet, millet, sorghum, rye, oats, pigeon oats, corn, buckwheat, amaranth, quinoa, etc.
- vegetables include, but are not limited to, pumpkin, carrot, radish, rutabaga, parsnip, turnip, black salsify, lotus root, beet (preferably beetroot: a variety improved for edible beet roots), water chestnut, shallot, garlic, scallion, lily of the valley, kale, onion, asparagus, udo, cabbage, lettuce, spinach, Chinese cabbage, rapeseed, komatsuna, bok choy, Chinese chives, green onions, Nozawana, butterbur, Swiss chard, mizuna, tomato, and eggplant.
- the vegetables preferred are those of the Solanum genus such as potato (particularly tubers), those of the Ipomoea genus such as sweet potato, and those of the Cucurbita genus such as pumpkin.
- the plant-based ingredients vegetables, potatoes, mushrooms, fruits, algae, grains (especially millet), nuts and seeds, etc. listed in the food group classification in the Standard Tables of Food Composition in Japan, 2015 Edition (7th revision), wild plants that are usually eaten as vegetables (plantain, bracken, butterbur, mugwort, etc.) can also be used.
- nuts and seeds include, but are not limited to, almonds, hemp, linseed, perilla, cashew nuts, pumpkin seeds, torreya, ginkgo nuts, chestnuts, walnuts, poppy seeds, coconuts, sesame seeds, Japanese chestnuts, horse chestnuts, lotus seeds, water chestnuts, pistachios, sunflower seeds, Brazil nuts, hazelnuts, pecans, macadamia nuts, pine trees, and peanuts.
- tubers include, but are not limited to, sweet potato, cassava, yacon, taro, taro, konjac, Polynesian arrowroot, potato, purple potato, Jerusalem artichoke, dogtooth violet, yam, Chinese yam, Chinese yam, and kudzu.
- mushrooms include, but are not limited to, shiitake mushrooms, matsutake mushrooms, wood ear mushrooms, maitake mushrooms, polyporus mushrooms, oyster mushrooms, king oyster mushrooms, enoki mushrooms, shimeji mushrooms, armillaria mushrooms, mushrooms, nameko mushrooms, amitake mushrooms, hattake mushrooms, and lactobacillus mushrooms.
- fruits include, but are not limited to, acerola, avocado, apricot, strawberry, fig, plum, citrus fruits (iyokan, satsuma mandarin, orange, grapefruit, lime, lemon, etc.), olive, persimmon, kiwi, guava, coconut, pomegranate, watermelon, plum, cherry (cherry, black cherry, etc.), jujube, pineapple, haskap, banana, papaya, loquat, grape, berry (blueberry, raspberry, etc.), mango, mangosteen, melon, peach, apple, etc.
- the edible plant contains starch at a predetermined rate or more.
- the starch content of the edible plant may be, for example, 4% by mass or more and 95% by mass or less.
- the effect of suppressing the fermented odor, grassy odor, and stringiness of the fermented food is further improved.
- the principle is not clear, it is possible that the effect is achieved by decomposing starch during the fermentation process and maintaining a certain rate or more of soluble carbohydrates.
- the lower limit of the starch content is preferably 4% by mass or more, more preferably 10% by mass or more, even more preferably 15% by mass or more, even more preferably 20% by mass or more, particularly preferably 25% by mass or more, and particularly preferably 30% by mass or more.
- the upper limit of the content is, for example, 95% by mass or less, preferably 90% by mass or less, more preferably 80% by mass or less, and even more preferably 70% by mass or less.
- the edible plant at least one selected from the group consisting of beans, grains, vegetables, potatoes, and nuts and seeds, which contain a certain percentage or more of starch, is preferred, with beans being particularly preferred.
- the starch content of beans, grains, vegetables, potatoes, and nuts and seeds may be, for example, 4% by mass or more and 95% by mass or less.
- the genus Maize, the genus Barley, and the genus Wheat are preferred.
- the genus Pisum such as peas
- the genus Chickpea such as chickpea
- Phaseolus are preferred.
- the vegetables the genus Solanum (such as eggplant), the genus Cucurbita (such as pumpkin), and the like are preferred, and as the potatoes, the genus Ipomoea (such as potato and sweet potato) are preferred.
- the starch content (mass%) of edible plants refers to the starch content in the edible plants before adjusting for the moisture content on a dry basis. If the moisture content on a dry basis is not adjusted, it refers to the starch content in the edible plants before fermentation.
- Starch content can be measured according to the method in the 2015 Edition (7th Edition) of the Standard Tables of Food Composition in Japan, using AOAC Method 996.11 (AOAC, 2005). It can also be determined more clearly by referring to the "starch” item in the 2015 Edition (7th Edition) of the Standard Tables of Food Composition in Japan.
- the edible plant contains palmitic acid (CAS registration number: 57-10-3) at a predetermined rate or more, and may be, for example, 1 ppm or more and 10,000,000 ppm or less.
- palmitic acid CAS registration number: 57-10-3
- methyl palmitate and ethyl palmitate are contained in the fermented food, which further improves the effect of suppressing the fermented odor and grassy odor of the fermented food.
- the content there is no particular lower limit on the content, but it is, for example, 1 ppm or more, preferably 10 ppm or more, more preferably 100 ppm or more, even more preferably 1000 ppm or more, even more preferably 1500 ppm or more, particularly preferably 2000 ppm or more, and particularly preferably 2500 ppm or more.
- the upper limit of the content is not particularly limited, but is, for example, 10,000,000 ppm or less, preferably 1,000,000 ppm or less, and more preferably 100,000 ppm or less.
- the palmitic acid content (ppm) of an edible plant represents the palmitic acid content in the edible plant before adjusting for dry weight moisture content. If the dry weight moisture content is not adjusted, it represents the palmitic acid content in the edible plant before fermentation.
- the palmitic acid content can be measured by flame ionization detection-gas chromatography of samples esterified after lipid extraction, following the method in the 2015 Edition (7th Edition) of the Standard Tables of Food Composition in Japan. A more clear determination can also be made by referring to the "palmitic acid" entry in the 2015 Edition (7th Edition) of the Standard Tables of Food Composition in Japan.
- the mass ratio of the methyl palmitate content (ppb) to the palmitic acid content (ppm) is a predetermined ratio or more.
- the mass ratio can be, for example, 0.000001 or more and 100 or less.
- the lower limit of the content there is no particular limit on the lower limit of the content, but it is, for example, 0.000001 or more, preferably 0.00001 or more, more preferably 0.0001 or more, even more preferably 0.001 or more, even more preferably 0.01 or more, particularly preferably 0.1 or more, and particularly preferably 1.0 or more.
- the upper limit of the content is not particularly limited, but is, for example, 100 or less, preferably 80 or less, more preferably 50 or less, even more preferably 10 or less, and even more preferably 1 or less.
- the mass ratio of the ethyl palmitate content (ppb) to the palmitic acid content (ppm) is a predetermined ratio or more.
- the mass ratio can be, for example, 0.000001 or more and 100 or less.
- the lower limit of the content there is no particular limit on the lower limit of the content, but it is, for example, 0.000001 or more, preferably 0.00001 or more, more preferably 0.0001 or more, even more preferably 0.001 or more, even more preferably 0.01 or more, particularly preferably 0.1 or more, and particularly preferably 1.0 or more.
- the upper limit of the content is not particularly limited, but is, for example, 100 or less, preferably 80 or less, more preferably 50 or less, even more preferably 10 or less, and even more preferably 1 or less.
- the fermented product of an edible plant is not particularly limited as long as it is made by fermenting an edible plant and is edible.
- a microorganism having ⁇ -amylase activity and a bacterium having the ability to produce PGA are particularly preferred, and it is more preferred to include a microorganism having ⁇ -amylase activity, and it is particularly preferred to include a microorganism having ⁇ -amylase activity and a bacterium having the ability to produce PGA ( ⁇ -polyglutamic acid).
- a microorganism having ⁇ -amylase activity there is no particular limitation, but koji is particularly preferred.
- bacteria having the ability to produce PGA there is no particular limitation, but bacteria of the genus Bacillus are preferred, and bacteria such as Bacillus subtilis and Bacillus subtilis belonging to Bacillus velezensis are more preferred, and Bacillus natto is particularly preferred from the viewpoint of imparting umami and richness to fermented foods.
- Koji is a general term for fermented products that are made by steaming grains or beans, then adding and propagating mold spores called seed koji.
- Any common grain or bean can be used as the raw material for making koji, with typical examples including rice, wheat, and soybeans.
- any mold used in general food brewing can be used as the seed koji fungus used to make koji, with molds of the genus Aspergillus being typical. More specifically, white koji mold (e.g., Aspergillus awamori var. kawachii, Aspergillus luchuensis mut. kawachii, Aspergillus usamii mut.
- shirousamii Aspergillus kawachii, etc.
- black koji mold e.g., Aspergillus awamori, Aspergillus niger, etc.
- yellow koji mold Alignid oryzae
- soy sauce koji mold Alignid ory sauce koji mold
- the requirement for koji is that it has enzyme activity, such as saccharifying enzymes, that convert polysaccharides such as starch contained in the raw materials into monosaccharides when used, and the live or dead state of the starter culture is not important.
- the koji in the present invention may be koji fungus in a vegetative cell state (so-called starter culture) that is normally used when producing soybean koji, or it may be koji in a spore state whose ⁇ -amylase activity has been increased to a predetermined level or more by koji production, but it is preferable to use koji in a spore state whose ⁇ -amylase activity has been increased to a predetermined level or more by koji production.
- the ⁇ -amylase activity of the koji has a predetermined activity, and may be, for example, 20 U/g or more and 100,000 U/g or less.
- the lower limit is not particularly limited, but may be 20 U/g or more, or 30 U/g or more, or 40 U/g or more, or 50 U/g or more, or 60 U/g or more, or 70 U/g or more, or 80 U/g or more, or 90 U/g or more, or 100 U/g or more, or 150 U/g or more, or 200 U/g or more, or 300 U/g or more, or 400 U/g or more, or 500 U/g or more, or 600 U/g or more, or 700 U/g or more, or 800 U/g or more, or 900 U/g or more, more preferably 1000 U/g or more, particularly preferably 1500 U/g or more, and particularly preferably 2000 U/g or more.
- the upper limit is not particularly limited, but is usually 100,000 U/g or less, preferably 80,000 U/g or less.
- ⁇ -Amylase activity can be measured, for example, according to the following method:
- ⁇ Activity measurement 2 mL of 0.05% soluble starch (Fujifilm Wako Pure Chemical Industries, Ltd., Starch (soluble) CAS9005-25-8, product code 195-03961) is placed in a test tube, and after standing at 37°C for 10 minutes, 0.25 mL of the enzyme solution is added and mixed. The mixture is then stood at 37°C for 30 minutes, and 0.25 mL of 1M HCl is added and mixed. Then, 0.25 mL of iodine potassium iodide solution containing 0.05 mol/L of iodine (0.05 mol/L iodine solution: Fujifilm Wako Pure Chemical Industries, Ltd.
- the iodine solution in the present invention refers to a diluted solution of iodine potassium iodide solution containing 0.05 mol/L of iodine (sometimes simply referred to as "0.05 mol/L iodine solution” or "0.05 mol/L iodine liquid" in the present invention).
- a mixed iodine potassium iodide solution of 93.7 mass% water, 0.24 mol/L (4.0 mass%) potassium iodide, and 0.05 mol/L (1.3 mass%) iodine ("0.05 mol/L iodine solution (product code 091-00475)" manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) is used by diluting it.
- a "0.25 mM iodine solution” can be obtained by diluting the "0.05 mol/L iodine solution" 200 times with water.
- Enzyme activity units (U / g): The rate of absorbance decrease C (%) during the enzyme reaction of the measurement sample for 30 minutes is calculated by the "rate of absorbance decrease in the enzyme reaction area (absorbance A) relative to the comparison area (absorbance B) ( ⁇ (absorbance B-absorbance A)/absorbance B ⁇ x 100 (%)".
- the enzyme activity that reduces absorbance by 10% per 10 minutes is defined as 1 unit (U), and the enzyme activity per 1 g of measurement sample is calculated from the rate of absorbance decrease C (%) when the enzyme reaction is carried out for 30 minutes using 0.25 mL of enzyme solution (sample content 0.025 g) using the following formula.
- koji making refers to the process of inoculating the steamed ingredients with koji mold, then leaving them at 30-40°C for at least 48 hours, going through the following steps: hikikomi ⁇ tokomomi ⁇ kirigaeshi ⁇ mori ⁇ nakashigoto ⁇ shimaigoto ⁇ dekoji.
- the gamma-polyglutamic acid (PGA) in the present invention is a polymer in which glutamic acid is polymerized through gamma-amide bonds. This substance is the main component of the stringy component (sticky component) of natto.
- the PGA that can be used in the present invention is not particularly limited, but can be obtained, for example, from a culture of bacteria of the genus Bacillus. Bacillus bacteria have the property of producing gamma-PGA during the culture process and releasing it outside the bacterial cell.
- bacteria such as natto bacteria belonging to Bacillus subtilis and Bacillus subtilis belonging to Bacillus velezensis are preferred, and natto bacteria are particularly preferred from the viewpoint of imparting umami and richness to fermented foods.
- natto bacteria can be used.
- common commercially available bacteria such as Miyagino bacteria (product name: pure cultured natto bacteria (Miyagino natto bacteria)) (manufactured by Miyagino Manufacturing Co., Ltd.), Takahashi bacteria (product name: Natto base) (manufactured by Takahashi Yuzo Laboratory Co., Ltd.), and Naruse bacteria (product name: powdered natto bacteria) (manufactured by Naruse Fermentation Chemistry Laboratory) can be used, but various strains such as mutant strains with specific properties and genetically modified strains can also be used.
- Natto bacteria are classified as Bacillus subtilis, but a variant of Bacillus subtilis, such as Bacillus subtilis var. It is a bacterium that is classified as Bacillus subtilis (natto) or Bacillus subtilis (natto) to distinguish it from Bacillus subtilis, or as Bacillus natto, a closely related species of Bacillus subtilis.
- the viable cell count of the bacteria capable of producing PGA is preferably 1.0 ⁇ 10 5 /g or more and 1.0 ⁇ 10 9 /g or less. More specifically, the lower limit is 1.0 ⁇ 10 5 /g or more, preferably 2.0 ⁇ 10 5 /g or more, more preferably 3.0 ⁇ 10 5 /g or more, and even more preferably 4.0 ⁇ 10 5 /g or more. On the other hand, the upper limit is 1.0 ⁇ 10 9 /g or less, preferably 8.0 ⁇ 10 8 /g or less, more preferably 6.0 ⁇ 10 8 /g or less, and even more preferably 4.0 ⁇ 10 8 /g or less.
- the viable cell count of bacteria capable of producing PGA can be measured as follows.
- the viable cell count in a fermented food can be measured, for example, by culturing a diluted solution of a natto suspension on an agar medium and counting the number of colonies.
- the number of colonies appearing other than natto bacteria can be suppressed by shortening the culture time (37°C for 18 hours) compared to the usual viable cell measurement (37°C for about 48 hours).
- the fermented food natto is placed in a filter-equipped bag attached to a paddle-type blender "Stomacher (registered trademark)", phosphate buffer is poured in, and the mixture is shaken with the Stomacher, diluted with phosphate buffer, and then mixed with a standard agar medium (manufactured by Atect Co., Ltd.), cultured at 37°C for 18 hours, and the number of colonies that appear there can be calculated.
- a standard agar medium manufactured by Atect Co., Ltd.
- the fermented food of the present invention preferably contains a certain percentage or more of soluble carbohydrates, and can contain, for example, 2.0% by mass or more and 50% by mass or less.
- soluble carbohydrates By setting the content of soluble carbohydrates in the fermented food within the above range, the effect of suppressing the fermented odor, grassy odor, and stringiness of the fermented food is further improved.
- the principle is unclear, it is possible that the above effect is achieved by decomposing starch during the fermentation process and maintaining a certain percentage or more of soluble carbohydrates.
- the lower limit of the content of soluble carbohydrates is, for example, 2.0% by mass or more, more preferably 4.0% by mass or more, even more preferably 5.0% by mass or more, even more preferably 7.0% by mass or more, particularly preferably 8.0% by mass or more, and particularly preferably 10% by mass or more.
- the upper limit of the content is not particularly limited, but is 50% by mass or less, preferably 40% by mass or less, more preferably 30% by mass or less, and even more preferably 25% by mass or less.
- soluble carbohydrates refers to carbohydrates that are soluble in water, and is a general term for monosaccharides and oligosaccharides (sugars in which about 2 to 10 monosaccharides are bonded). Therefore, starch, which is a component in which far more sugars are bonded, is not included in the concept.
- the soluble carbohydrate content can be determined by adding up each measured value obtained by comparing the content with a standard monosaccharide or oligosaccharide (2 to 10 sugars) of known concentration using high performance liquid chromatography in accordance with the measurement method for "available carbohydrates (glucose, fructose, galactose, sucrose, maltose, lactose and trehalose)" in the "Analysis Manual for the 2015 Edition (7th Edition) of the Standard Tables of Food Composition in Japan.”
- the "average soluble carbohydrate content” refers to the average value of the soluble carbohydrate content of the composition in the process of fermentation with bacteria capable of producing PGA (particularly Bacillus subtilis natto).
- the average soluble carbohydrate content does not mean that the soluble carbohydrate content does not completely fall outside the range of the soluble carbohydrate content, and even if the soluble carbohydrate content falls outside the range of the specified soluble carbohydrate content for a short period of time (for example, within 40 minutes, preferably within 20 minutes), the average soluble carbohydrate content can be satisfied.
- the soluble carbohydrate content in the composition can be increased by the metabolism of the microorganisms or by adding soluble carbohydrates, so that the average soluble carbohydrate content falls within the specified range.
- the average soluble carbohydrate content may be adjusted based on soluble carbohydrates contained in food ingredients such as edible plants that are the raw materials for the fermented food, or may be added separately from the food ingredients, or may be generated during the production of the fermented food of the present invention, or may be generated during the fermentation of bacteria capable of producing soluble carbohydrates, or a combination thereof.
- the average soluble carbohydrate content may be adjusted by including bacteria capable of producing soluble carbohydrates (e.g., koji) in addition to bacteria capable of producing PGA.
- bacteria capable of producing soluble carbohydrates e.g., koji
- the average soluble carbohydrate content may be adjusted by including bacteria capable of producing soluble carbohydrates (e.g., koji) in addition to bacteria capable of producing PGA.
- the fermented food of the present invention can contain a predetermined proportion of starch.
- Starch includes starch derived from edible plants that are the raw material of the fermented food.
- the starch content in the fermented food of the present invention may be, for example, 2% by mass or more and 30% by mass or less.
- the lower limit of the starch content is preferably 2% by mass or more, more preferably 3% by mass or more, even more preferably 4% by mass or more, even more preferably 6% by mass or more, particularly preferably 8% by mass or more, and particularly preferably 10% by mass or more.
- the upper limit of the content is preferably 30% by mass or less, more preferably 25% by mass or less, even more preferably 20% by mass or less, and even more preferably 15% by mass or less.
- the starch content (mass%) of fermented foods refers to the starch content in the fermented foods before adjusting for moisture content on a dry basis.
- Starch content can be measured according to the method in the 2015 edition (7th revision) of the Standard Tables of Food Composition in Japan, using AOAC method 996.11 (AOAC, 2005). It can also be determined more clearly by referring to the "starch" section in the 2015 edition (7th revision) of the Standard Tables of Food Composition in Japan.
- the mass ratio of soluble carbohydrates to starch is preferably a predetermined ratio or more.
- the mass ratio can be, for example, 0.05 or more and 10 or less.
- the lower limit of the mass ratio is preferably 0.05 or more, more preferably 0.1 or more, even more preferably 0.2 or more, even more preferably 0.4 or more, particularly preferably 0.6 or more, and especially more preferably 0.8 or more.
- the upper limit of the mass ratio is preferably 10 or less, more preferably 8 or less, and even more preferably 6 or less.
- the fermented food of the present invention preferably contains ⁇ -amylase (CAS registration number: 9000-90-2) having a certain level of activity or higher.
- ⁇ -amylase is an enzyme that irregularly cleaves the ⁇ -1,4-bonds of starch to produce polysaccharides, maltose, and oligosaccharides.
- the ⁇ -amylase may be contained in the food material such as an edible plant that is the raw material for the fermented food of the present invention, may be added separately from the food material, may be generated during the production of the fermented food of the present invention, or may be a combination of these.
- purified ⁇ -amylase may be used, a food material containing ⁇ -amylase may be used, or a microorganism having ⁇ -amylase activity may be used, but it is preferable to use a microorganism having ⁇ -amylase activity, and it is preferable to use koji as the microorganism having ⁇ -amylase activity.
- the fermented food of the present invention preferably has a predetermined ⁇ -amylase activity, and may be, for example, 2 U/g or more and 100,000 U/g or less.
- the lower limit is not particularly limited, but is 2 U/g or more, or 3 U/g or more, or 4 U/g or more, or 5 U/g or more, or 6 U/g or more, or 7 U/g or more, or 8 U/g or more, or 9 U/g or more, or 10 U/g or more, or 15 U/g or more, or 20 U/g or more, or 30 U/g or more, or 40 U/g or more, or 50 U/g or more, or 60 U/g or more, or 70 U/g or more, or 80 U/g or more, or 90 U/g or more, more preferably 100 U/g or more, particularly preferably 150 U/g or more, and particularly preferably 200 U/g or more.
- the upper limit is not particularly limited, but is usually 100,000 U/g or less, preferably
- ⁇ -Amylase activity can be measured and calculated, for example, according to the following method.
- ⁇ Activity measurement 2 mL of 0.05% soluble starch (Fujifilm Wako Pure Chemical Industries, Ltd., Starch (soluble) CAS9005-25-8, product code 195-03961) is placed in a test tube, and after standing at 37°C for 10 minutes, 0.25 mL of the enzyme solution is added and mixed. The mixture is then stood at 37°C for 30 minutes, and 0.25 mL of 1M HCl is added and mixed. Then, 0.25 mL of iodine potassium iodide solution containing 0.05 mol/L of iodine (0.05 mol/L iodine solution: Fujifilm Wako Pure Chemical Industries, Ltd.
- the iodine solution in the present invention refers to a diluted solution of iodine potassium iodide solution containing 0.05 mol/L of iodine (sometimes simply referred to as "0.05 mol/L iodine solution” or "0.05 mol/L iodine liquid" in the present invention).
- a mixed iodine potassium iodide solution of 93.7 mass% water, 0.24 mol/L (4.0 mass%) potassium iodide, and 0.05 mol/L (1.3 mass%) iodine ("0.05 mol/L iodine solution (product code 091-00475)" manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) is used by diluting it.
- a "0.25 mM iodine solution” can be obtained by diluting the "0.05 mol/L iodine solution" 200 times with water.
- Enzyme activity units (U / g): The rate of absorbance decrease C (%) during the enzyme reaction of the measurement sample for 30 minutes is calculated by the "rate of absorbance decrease in the enzyme reaction area (absorbance A) relative to the comparison area (absorbance B) ( ⁇ (absorbance B-absorbance A)/absorbance B ⁇ x 100 (%)".
- the enzyme activity that reduces absorbance by 10% per 10 minutes is defined as 1 unit (U), and the enzyme activity per 1 g of measurement sample is calculated from the rate of absorbance decrease C (%) when the enzyme reaction is carried out for 30 minutes using 0.25 mL of enzyme solution (sample content 0.025 g) using the following formula.
- the fermented food of the present invention may have a value N calculated by the following formula of less than 1.0.
- N number ⁇ ⁇ number ⁇ Value
- the numerical value N being less than 1.0 is preferable since it has the effect of suppressing stringiness.
- the upper limit is less than 1.0, preferably 0.5 or less, more preferably 0.3 or less, and even more preferably 0.2 or less.
- the lower limit is not particularly limited, but may be, for example, 0.001 or more, 0.005 or more, 0.01 or more, 0.02 or more, 0.05 or more, or 0.1 or more.
- PGA and levan are components of the stringiness component (stickiness component) of natto, and it is believed that the interaction between these components affects the strength and stabilization of stringiness of natto, so it is believed that the stringiness of natto is suppressed by having the numerical value N be less than the specified range.
- the PGA content can be measured, for example, according to the following method.
- ⁇ Pretreatment for PGA analysis> For samples used in PGA analysis of fermented foods, the viscous substance extract of the fermented food obtained by carrying out the protein removal treatment described below and then purification treatment using ethanol is used as the sample. 1. 10 g of the fermented food is added to 50 mL of 2.5% trichloroacetic acid (hereinafter also referred to as TCA), heated to 50° C. for 10 minutes, and stirred. 2. After removing the solid portion from the mixture, the mixture is centrifuged (12,000 rpm, 10 min) to obtain the supernatant. 3.
- TCA trichloroacetic acid
- the obtained supernatant is adjusted to pH 7.0 with sodium hydroxide and diluted two-fold with ion-exchanged water, and then an equal amount of ethanol that has been pre-cooled to -30°C is added to the neutralized liquid and stirred. 4. After leaving the mixture on ice for 10 minutes, it is centrifuged (12,000 rpm, 10 min), the supernatant is discarded, and the mixture is dried. 5. The precipitate obtained by drying is used as a viscous substance, and the solution is dissolved in 20 mM phosphate buffer (pH 7.0) to be used as a viscous substance extract.
- PGA in the fermented food was measured by measuring the absorbance of the viscous extract that had been pretreated as described above and the solution that had been treated in the following manner, and comparing the absorbance with that of a standard substance to perform a quantitative analysis.
- the absorbance was measured using a UV-1800 UV-visible spectrophotometer (Shimadzu Corporation). 1.
- To prepare a standard calibration curve prepare standard solutions of 0, 25, 50, and 100 ⁇ g/mL of "poly- ⁇ -glutamic acid" (average molecular weight 1,500,000 to 2,500,000) (FUJIFILM Wako Pure Chemical Corporation) diluted in 20 mM phosphate buffer (pH 7.0) as PGA standard solutions. 2.
- the viscous substance extract obtained by the above method is diluted with 20 mM phosphate buffer (pH 7.0) so that it falls within the concentration range of the calibration curve. 3.
- 20 mM phosphate buffer (pH 7.0) To 0.5 mL of the PGA standard solution and the analytical sample, add 2.0 mL of 20 mM phosphate buffer (pH 7.0), add 0.5 mL of 0.1 M cetylmethyltrimethylammonium bromide (Cetablon)/1 M NaCl solution, and stir. 4.
- the Abs (absorbance) at a wavelength of 400 nm is measured, and the PGA concentration of the sample is calculated using a calibration curve prepared from the standard solution.
- Levan is a polymer of fructose that is widely found in nature and is produced by many microorganisms and plants, and is one of the components that make up the sticky substance in natto.
- the levan content can be measured according to the following method.
- Pretreatment for levan analysis> For samples used in levan analysis of fermented foods, the viscous substance extract of the fermented food obtained by carrying out the protein removal treatment described below and then purification treatment using ethanol is used as the sample. 1. 10 g of the fermented food is added to 50 mL of 2.5% trichloroacetic acid (hereinafter also referred to as TCA), heated to 50° C. for 10 minutes, and stirred. 2. After removing the solid portion from the mixture, the mixture is centrifuged (12,000 rpm, 10 min) to obtain the supernatant. 3.
- TCA trichloroacetic acid
- the obtained supernatant is adjusted to pH 7.0 with sodium hydroxide and diluted two-fold with ion-exchanged water, and then an equal amount of ethanol that has been pre-cooled to -30°C is added to the neutralized liquid and stirred. 4. After leaving the mixture on ice for 10 minutes, it is centrifuged (12,000 rpm, 10 min), the supernatant is discarded, and the mixture is dried. 5. The precipitate obtained by drying is used as a viscous substance, and the solution is dissolved in 20 mM phosphate buffer (pH 7.0) to be used as a viscous substance extract.
- the amount of levan in fermented foods is measured by measuring the absorbance of the extract of the viscous substance that has been pretreated as described above and the solution that has been treated in the following manner, and then quantitative analysis is performed by comparing the absorbance with that of a standard substance that uses fructose as an index.
- a UV-1800 ultraviolet-visible spectrophotometer (Shimadzu Corporation) is used to measure the absorbance. 1. To prepare a standard calibration curve, prepare fructose standard solutions of 0, 10, 25, 50, and 100 ⁇ g/mL in 20 mM phosphate buffer (pH 7.0). 2.
- the thickening substance extract obtained above is diluted with 20 mM phosphate buffer (pH 7.0) according to the concentration so that it falls within the range of the above calibration curve.
- 3. 0.6 mL of the fructose standard solution and the analytical sample were placed in a test tube containing 0.3 mL of resorcinol-thiourea reagent, and 2.1 mL of 30% HCl were gently mixed. The mixture was covered with a marble or similar to prevent bumping, and incubated at 80°C for 10 minutes.
- Abs (absorbance) was measured at a wavelength of 500 nm, and the levan concentration was calculated using a calibration curve prepared from the standard solution. The levan concentration was calculated by subtracting the water in the binding portion and multiplying the fructose calibration curve by 0.9.
- the fermented food of the present invention is preferably such that the viscosity of the extract obtained by stirring 40 g of the fermented food in 40 mL of water and then removing the solid portion is a predetermined viscosity or less under the conditions of a B-type viscometer (60 rpm, 25°C, rotor No. 2).
- the viscosity may be, for example, 180 mPa ⁇ s or less.
- the upper limit of the viscosity under the above conditions is preferably 180 mPa ⁇ s or less, more preferably 170 mPa ⁇ s or less, and even more preferably 160 mPa ⁇ s or less.
- the fermented food of the present invention preferably has a spinnability within a specified range. Specifically, when two grains of the fermented food sample are pressed tightly together and one grain is then pulled upward at a speed of 100 mm/min, the distance at which the thread formed between the two grains of the fermented food breaks is preferably 100 mm or less, more preferably 50 mm or less, even more preferably 25 mm or less, even more preferably 15 mm or less, particularly preferably 10 mm or less, and especially preferably 5 mm or less. On the other hand, there is no particular lower limit.
- a device capable of pulling up the sample at a constant speed can be used to measure spinnability, and as an example, a device combining a digital force gauge (model number FGP-0.5 (manufactured by Nidec-Shimpo Corporation)) and a force gauge stand (model number FGS-50E (manufactured by Nidec-Shimpo Corporation)) can be used.
- a digital force gauge model number FGP-0.5 (manufactured by Nidec-Shimpo Corporation)
- a force gauge stand model number FGS-50E (manufactured by Nidec-Shimpo Corporation)
- the moisture content of the fermented food of the present invention on a dry basis is preferably 30% by mass or more and 85% by mass or less from the viewpoint of storage stability and processability.
- the lower limit of the moisture content is preferably 35% by mass or more, more preferably 40% by mass or more, and even more preferably 45% by mass or more.
- the upper limit of the moisture content is preferably 80% by mass or less, more preferably 70% by mass or less, and even more preferably 60% by mass or less.
- the form of the fermented food of the present invention is not particularly limited, and can be, for example, a solid, paste, liquid, or gel.
- the form of the fermented food of the present invention is preferably solid when it is fermented using only koji. More specifically, it can be in a form in which the shape of the raw materials remains intact, such as soybean natto, in which the raw materials are crushed, such as crushed natto, in which the raw materials are crushed, or in a dried form.
- the fermented food of the present invention is suitable for eating as is after fermentation.
- fermented foods fermented with Bacillus subtilis natto and Aspergillus oryzae there are no particular limitations on their form.
- they can be in a solid, paste, liquid, or gel form. More specifically, they can be in a form in which the shape of the raw material remains intact, such as soybean natto, or in which the raw material has been cracked, crushed, or ground, or in a dried form.
- the fermented food of the present invention is suitable for eating as is after fermentation.
- the fermented food of the present invention can also be processed and used. There are no particular limitations on the type of food that is the subject of the present invention, as long as it is in a form that encompasses the fermented food of the present invention.
- the fermented food of the present invention has a good texture and sweetness, and is therefore preferably used, for example, in seasonings, desserts, baked goods, steamed goods, frozen desserts, chocolate confectionery, gum, candy, gummy candy, cream, jam, processed foods, cooked foods, retort foods, or cereal foods, solid foods for ingesting vegetables, dosage-type foods (health and nutritional (supplement) foods), animal feed, etc.
- the value N is less than 1.0, and/or the viscosity of the extract obtained by stirring 40 g of the fermented food in 40 mL of water and then removing the solid portion is 180 mPa ⁇ s or less under the conditions of a B-type viscometer (60 rpm, 25°C, rotor No.
- the distance until the thread formed between the two grains of the fermented food breaks is 100 mm or less, so that the fermented food of the present invention can be easily provided to the animal that ingests it by the feeder.
- the fermented food of the present invention can be provided without soiling the hair or body of the animal that ingests it, or the hands of the feeder.
- Examples of animals that ingest the fermented food of the present invention include pet animals such as canines and felines, and livestock (cows, pigs, chickens, etc.), and pet animals are preferable, and canines are more preferable because they prefer a certain degree of fermentation odor.
- the food also includes foods that are cooked so that they can be eaten at home. More specifically, for example, the fermented food may be baked or boiled, crushed or ground, dried, or powdered. Furthermore, the fermented food of the present invention may be mixed with other foods (including fermented and unfermented foods) and seasonings, and then eaten, or processed or cooked. This can impart an appropriate sweetness and texture to the cooked product. In addition, sterilization or low-temperature distribution may be performed to suppress excessive fermentation by koji or natto bacteria.
- the fermented food of the present invention may be processed as a paste or liquid food or drink, or may be added to other foods and drinks to be consumed. This can improve the overall taste (flavor and texture) of the food or drink, or enhance the taste of the other foods and drinks.
- processed grain products include processed grain products. Processed grain products refer to foods and drinks produced by processing the above-mentioned grains, and specific examples include cooked rice, noodles, bread, waffles, cereals, beverages, etc.
- Cod rice refers to processed cooked rice products made by adding water to rice and boiling or steaming it, and examples of such products include white rice, salted rice, red rice, sticky rice, cooked rice, mixed rice, rice balls, sushi rice, mochi, and dumplings.
- rice include non-glutinous rice, glutinous rice, pre-washed rice with different degrees of polishing, and brown rice.
- noodles are foods made from wheat flour, rice flour, buckwheat flour, beans, or other cereal flour, which are formed and processed into noodle, plate, ribbon, or other shapes and then cooked by boiling, stewing, or steaming.
- Examples include soba, udon, kishimen, ramen, Chinese noodles, pasta, macaroni, somen, pho, Korean hiyashimen, and vermicelli.
- "Breads” are foods made from wheat flour, rice flour, buckwheat flour, beans, or other cereal flour, which are added with yeast, or which are kneaded with water, salt, fruit, vegetables, eggs, and their processed products, sugar, edible oils, and other ingredients, fermented, and then baked, with a moisture content of 10% or more.
- “Waffles” are foods made from wheat flour, rice flour, buckwheat flour, beans, or other cereal flour, which are mixed with eggs, butter, milk, sugar, and other ingredients, and then sandwiched between two iron plates and baked.
- “Cereals” are foods made by baking and processing grains such as corn, wheat, oats, and rice (brown rice). By adding the fermented food of the present invention, the deliciousness (flavor and texture) of the processed grain products
- the present invention relates to a method for producing a fermented food of the present invention (also referred to as the "production method of the present invention” in this specification), which comprises a step of fermenting (I) a composition (sometimes referred to as the "composition of the present invention” in this specification) that contains at least one edible plant selected from the group consisting of nuts and seeds, grains, beans, vegetables, potatoes, mushrooms, and fruits, and koji, and has a salt content of 1000 mg or less per 100 g, at a product temperature of 30° C. or higher for 5 hours or more. This will be described below.
- koji Any type of koji may be used.
- the only requirement is that the koji has enzyme activity, such as a saccharifying enzyme, that converts starch and other polysaccharides contained in the raw materials into saccharification when used, and the live or dead state of the starter culture is not important.
- the edible plants and koji may be mixed at any time before the start of the fermentation process described below.
- the koji blended in the composition of the present invention preferably has a moisture content on a dry basis of a predetermined percentage or less, and may be, for example, 0.1% by mass or more and 50% by mass or less.
- a moisture content on a dry basis of a predetermined percentage or less By making the moisture content of the koji on a dry basis of a predetermined percentage or less, the effect of the present invention of suppressing the fermented odor, grassy odor, and stringiness of the fermented food is further improved.
- the principle is not clear, it is thought that when the difference in the moisture content on a dry basis of the koji and the edible plant becomes a certain amount or more, the koji adheres to the surface of the edible plant, and fermentation proceeds in the edible plant that has moisture.
- the upper limit of the moisture content is 50% by mass or less, preferably 40% by mass or less, more preferably 30% by mass or less, even more preferably 20% by mass or less, even more preferably 15% by mass or less, particularly preferably 10% by mass or less, and especially more preferably 5% by mass or less.
- the lower limit is not particularly limited, but is, for example, 0.1% by mass or more, 0.5% by mass or more, or 1% by mass or more.
- the d50 after ultrasonic treatment refers to the particle diameter at which the cumulative particle frequency percentages on the larger side and the smaller side are equal when a particle diameter distribution obtained by measurement using a laser diffraction particle size distribution analyzer and ethanol as the measurement solvent is divided into two at a certain particle diameter, and is also expressed as "d50".
- particle diameters are all measured on a volume basis, and unless otherwise specified, the measured particle diameters represent the results obtained by analyzing samples after ultrasonic treatment.
- “ultrasonic treatment” refers to a process in which ultrasonic waves with a frequency of 30 kHz and an output of 40 W are applied to the measurement sample in the measurement solvent for 3 minutes.
- the koji to be blended in the composition of the present invention is preferably in powder form (sometimes simply referred to as “koji powder” in the present invention), and its d50 after ultrasonic treatment (sometimes simply referred to as "average particle size” in the present invention) is preferably within a specified range.
- koji powder in powder form
- average particle size average particle size
- the d50 after ultrasonic treatment of the koji may be, for example, 0.1 ⁇ m or more and 1000 ⁇ m or less.
- the upper limit is 1000 ⁇ m or less, preferably 750 ⁇ m or less, more preferably 500 ⁇ m or less, even more preferably 400 ⁇ m or less, and particularly preferably 350 ⁇ m or less.
- the lower limit is not particularly specified, but from the viewpoint of industrial convenience, it is preferably 0.1 ⁇ m or more, or 1 ⁇ m or more.
- the particle size after ultrasonic treatment can be measured using ethanol as a solvent according to the procedure described below.
- the d50 of the koji powder after ultrasonic treatment can be adjusted to the above range using a sieve with appropriate mesh size. Specifically, by passing the koji powder through a mesh with mesh size corresponding to the upper limit size of the ultrasonic treatment d50 and optionally collecting the koji powder that passes through a mesh with mesh size corresponding to the lower limit size, it is possible to obtain koji powder with a d50 after ultrasonic treatment within a certain range.
- “mesh on” refers to the koji powder fraction that remains on a sieve of a particular size
- “mesh pass” refers to the koji powder fraction that passes through a sieve of a particular size.
- the content of each fraction is measured by fractionating the koji powder through sieves with different mesh sizes.
- “50 mesh on” means the koji powder fraction that remains on a 50 mesh sieve
- "0.1 mesh pass 50 mesh on” means the koji powder fraction that passes through a 0.1 mesh sieve and remains on a 50 mesh sieve.
- “mesh” is a unit that indicates the density of the meshes of wire mesh, sieves, filters, etc., and represents the number of meshes per inch.
- “1 mesh on (pass)” means the koji powder fraction that remains on (passes through) a sieve with an opening of 2.50 centimeters
- “0.1 mesh on (pass)” means the koji powder fraction that remains on (passes through) a sieve with an opening of 25.0 centimeters
- “50 mesh on (pass)” means the koji powder fraction that remains on (passes through) a sieve with an opening of 300 micrometers.
- the wire thickness and mesh spacing of the mesh-on are the values specified in the U.S.A. Standard Testing Sieves ASTM Specifications E 11-04 (for example, 50 mesh is the “Al (corresponding to "No. 50” in the “Standard for the 1990 Int. Inertial” and 1 mesh corresponds to "1.00") or a similar value is used, and 100 g of a sample (20°C) containing the koji powder to be measured is evenly spread on sieves stacked from the top in order from the largest mesh size to the smallest mesh size, and the size can be measured by processing while vibrating with a load that does not change the composition size until the weight of the fraction on each sieve becomes constant.
- the size of the koji powder fraction of the present invention can be, for example, in the range of 50 mesh on to 0.1 mesh pass.
- the lower limit is usually 50 mesh on, 42 mesh on, further 36 mesh on, further 30 mesh on, further 26 mesh on, further 22 mesh on, and particularly preferably 18 mesh on.
- the upper limit of the size of the koji powder of the present invention is not particularly limited, but is usually 0.1 mesh pass, and preferably 0.5 mesh pass, and particularly preferably 1 mesh pass.
- the edible plants to be incorporated into the composition of the present invention may be used raw, or may be dried (dried products).
- raw materials may be used that have been mechanically processed, such as crushed or cut, or chemically processed, such as dried or solution-treated.
- the edible plants to be incorporated in the composition of the present invention are preferably edible plants that have been heat-treated at 80°C or higher under conditions of a dry weight moisture content of 50% by mass or more. If the dry weight moisture content of the edible plant is low, a process of adjusting the moisture content to the aforementioned value by employing a hydration treatment such as soaking can be employed.
- a hydration treatment such as soaking
- There are no particular limitations on the heat treatment method but submerged heat treatments such as boiling or steaming are preferred. Among these, steaming is particularly preferred from the viewpoint of preventing loss of components. Note that it is desirable to soak the raw material in water and swell it before use, prior to the submerged heat treatment.
- a specific procedure for steaming edible plants can be, for example, a method in which the edible plants are soaked in water at 20°C or less for around 24 hours, then drained and steamed with steam at 100-135°C for 30 minutes. At this time, a method in which the plants are steamed under pressure, for example at a high pressure of 0.10-0.22 MPa, can also be used.
- the steaming process does not have to be carried out in one go, but can be divided into multiple steps by depressurizing and then repressurizing. Water can also be added after each steaming process.
- Specific preparation procedures for boiled ingredients include, for example, soaking the ingredients in water at 20°C or less for about 24 hours, and then boiling them in water at 90-100°C for 20-50 minutes.
- the composition of the present invention is obtained by mixing a predetermined amount of edible plant and koji (and, if necessary, other food ingredients).
- the mass ratio of koji to edible plant may be 0.1 or more and 0.3 or less. More specifically, the lower limit of the mass ratio is 0.1 or more, preferably 0.15 or more. Meanwhile, the upper limit is 0.3 or less, preferably 0.25 or less.
- composition of the present invention may be in the form of a liquid, sol, gel, or solid, as long as the food material is partially or completely integrated with water.
- the salt in the present invention refers to the total salt content contained in the composition, i.e., the total salt content including not only the salt mixed in the composition during preparation, but also the salt content contained in the food raw material and other optional components.
- the composition of the present invention preferably has a salt content of a predetermined ratio or less, and may be, for example, 1 mg or more and 1000 mg or less per 100 g. By making the salt content of the composition a predetermined ratio or less, fermentation can be efficiently promoted.
- the upper limit of the salt content is 1000 mg or less, preferably 500 mg or less, more preferably 300 mg or less, even more preferably 100 mg or less, and particularly preferably 50 mg or less per 100 g of the composition of the present invention.
- the lower limit is not particularly limited, but is, for example, 1 mg or more, preferably 2 mg or more, and more preferably 3 mg or more.
- the salt content can be measured, for example, according to the following method.
- acid decomposition is performed as follows: 10 ml of nitric acid is added to 0.5 g of a sample (pre-fermentation composition), and then heat treatment is performed under the following conditions using an acid decomposition apparatus Digiprep (manufactured by GL Sciences). (Conditions) Heat up (25 minutes) ⁇ Hold at 60°C (30 minutes) ⁇ Heat up (25 minutes) ⁇ Hold at 105°C (60 minutes).
- the acid digested solution after the above treatment is diluted with 1% nitric acid to 5,000 times the volume of the sample, and then subjected to sodium analysis using an ICP-MS 7000 (Agilent).
- the measured sodium content is converted to salt equivalent (2.54 times) to calculate the salt content.
- the composition of the present invention preferably contains a predetermined proportion of starch, for example, 4% by mass or more and 95% by mass or less.
- the lower limit of the starch content is 4% by mass or more, more preferably 10% by mass or more, even more preferably 15% by mass or more, even more preferably 20% by mass or more, particularly preferably 25% by mass or more, and especially preferably 30% by mass or more.
- the upper limit of the content is 95% by mass or less, preferably 90% by mass or less, more preferably 80% by mass or less, even more preferably 70% by mass or less, even more preferably 60% by mass or less, particularly preferably 50% by mass or less, and especially preferably 40% by mass or less.
- the starch may be contained in the foodstuff such as an edible plant that is the raw material of the fermented food of the present invention, may be added separately from the foodstuff, or may be a combination thereof, but is preferably derived from an edible plant.
- Starch content can be measured according to the method of the Standard Tables of Food Composition in Japan, 2015 Edition (7th revision), using AOAC method 996.11 (AOAC, 2005).
- the composition of the present invention preferably has a certain ⁇ -amylase activity, which may be, for example, 2 U/g or more and 20,000 U/g or less.
- the lower limit is not particularly limited, but is preferably 2 U/g or more, more preferably 10 U/g or more, and even more preferably 15 U/g or more.
- the upper limit is not particularly limited, but is usually 20,000 U/g or less, preferably 5,000 U/g or less, more preferably 1,000 U/g or less, and even more preferably 500 U/g or less.
- the ⁇ -amylase activity may be contained in the foodstuffs such as edible plants that are the raw materials for the fermented food of the present invention, or may be added separately from the foodstuffs, or may be a combination of these, but is preferably derived from microorganisms (particularly koji).
- ⁇ -Amylase activity can be measured and calculated, for example, according to the following method.
- ⁇ Activity measurement 2 mL of 0.05% soluble starch (Fujifilm Wako Pure Chemical Industries, Starch (soluble) CAS9005-25-8, product code 195-03961) is placed in a test tube, and after standing at 37°C for 10 minutes, 0.25 mL of the enzyme solution is added and mixed. The mixture is then stood at 37°C for 30 minutes, and 0.25 mL of 1M HCl is added and mixed. Then, 0.25 mL of iodine potassium iodide solution containing 0.05 mol/L of iodine (0.05 mol/L iodine solution: Fujifilm Wako Pure Chemical Industries, Ltd.
- the iodine solution in the present invention refers to a diluted solution of iodine potassium iodide solution containing 0.05 mol/L of iodine (sometimes simply referred to as "0.05 mol/L iodine solution” or "0.05 mol/L iodine liquid” in the present invention).
- a mixed iodine potassium iodide solution of 93.7 mass% water, 0.24 mol/L (4.0 mass%) potassium iodide, and 0.05 mol/L (1.3 mass%) iodine ("0.05 mol/L iodine solution (product code 091-00475)" manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) is used by diluting it.
- a "0.25 mM iodine solution” can be obtained by diluting the "0.05 mol/L iodine solution" 200 times with water.
- Enzyme activity units (U / g): The rate of absorbance decrease C (%) during the enzyme reaction of the measurement sample for 30 minutes is calculated by the "rate of absorbance decrease in the enzyme reaction area (absorbance A) relative to the comparison area (absorbance B) ( ⁇ (absorbance B-absorbance A)/absorbance B ⁇ x 100 (%)".
- the enzyme activity that reduces absorbance by 10% per 10 minutes is defined as 1 unit (U), and the enzyme activity per 1 g of measurement sample is calculated from the rate of absorbance decrease C (%) when the enzyme reaction is carried out for 30 minutes using 0.25 mL of enzyme solution (sample content 0.025 g) using the following formula.
- the dry moisture content of the composition of the present invention is preferably a predetermined value or more, and may be, for example, 20% by mass or more and 90% by mass or less.
- the upper limit of the dry moisture content is not limited, but is, for example, 90% by mass or less, preferably 80% by mass or less, and more preferably 70% by mass or less.
- the lower limit is not particularly limited, but can be usually 20% by mass or more, preferably 30% by mass or more, more preferably 40% by mass or more, and even more preferably 50% by mass or more.
- the water in the composition of the present invention may be derived from various raw materials of the composition, or may be derived from water added.
- the dry moisture content contained in the composition when the dry moisture content contained in the composition is low, a process of adjusting it to the above-mentioned value by adopting a hydration treatment such as immersion can be adopted.
- a specific preparation procedure for the dry moisture content for example, a process of immersing the raw materials in water of less than 10°C for about 24 hours can be adopted. If the water temperature during immersion is high, there is a possibility of contamination by bacteria, so it is preferable to immerse at 4°C.
- the production method of the present invention preferably includes (II) a step of fermenting with bacteria capable of producing PGA. More specifically, for example, it is preferable to include a step of blending bacteria capable of producing PGA with the composition of the present invention.
- bacteria capable of producing PGA there are no particular limitations on the bacteria capable of producing PGA, but bacteria of the genus Bacillus can be used. In particular, bacteria such as Bacillus subtilis belonging to natto and Bacillus subtilis belonging to Bacillus velezensis are more preferred, with Bacillus natto being even more preferred.
- the state of the natto bacteria to be added as the natto bacteria starter is not particularly limited, but it is preferable to use one in a spore state that can be directly inoculated into the high-temperature raw material to prevent contamination with other bacteria.
- the spore suspension can be a culture solution obtained by culturing the above-mentioned Bacillus subtilis natto in a liquid medium containing components suitable for spore formation.
- the components of the liquid medium are not particularly limited, so long as they enable spore formation and growth of Bacillus subtilis natto and contain medium components such as a carbon source, a nitrogen source, and inorganic salts that are commonly used in culturing Bacillus subtilis natto, and may be either a synthetic medium or a natural medium.
- carbon sources include sugars such as glucose, sucrose, galactose, mannose, starch, and starch hydrolysates, and organic acids such as citric acid
- nitrogen sources include peptone, meat extract, casein hydrolysate, ammonia, ammonium sulfate, and ammonium chloride
- inorganic salts include sodium chloride, potassium chloride, calcium chloride, sodium sulfate, sodium hydrogen sulfate, sodium nitrate, potassium phosphate, ferric chloride hexahydrate, magnesium sulfate heptahydrate, manganese chloride tetrahydrate, and ferrous sulfate.
- the medium may also contain yeast extract, malt extract, soybean flour, vitamins (biotin, etc.), etc.
- yeast extract malt extract
- soybean flour vitamins (biotin, etc.), etc.
- the number of natto bacteria to be inoculated is not particularly limited as long as the bacterial concentration is in accordance with the usual method for soybean natto, but is usually 1.0 x 10 to 1.0 x 10 per 1 g of steamed or boiled raw material.
- the product temperature of the raw material when inoculated is not particularly limited, but it is preferable to inoculate at a high temperature of about 55 to 95°C to prevent contamination with various bacteria during inoculation.
- the normal fermentation temperature range refers to a product temperature range of 30 to 60°C. If the fermentation temperature is lower than the specified range, the fermented food will not have a good flavor and it will be difficult to keep certain components within the specified range. If the fermentation temperature is higher than the specified range, excessive amounts of ammonia, lower fatty acids, etc. will be produced, which is undesirable.
- “maintaining the raw material temperature substantially in the normal fermentation temperature range for a predetermined time from the start of fermentation” does not mean that the temperature range is not exceeded completely, but means that the fermentation conditions are met even if the temperature is outside the range within a slight temperature range (e.g., within 3°C, preferably within 2°C) and for a short time (e.g., within 20 minutes, preferably within 10 minutes). Therefore, “average fermentation temperature” means the average value of the fermentation temperature in the process of fermentation with bacteria capable of producing PGA (especially natto bacteria).
- the average fermentation temperature is more than 40° C. and less than 65° C. when the number of bacteria is 1.0 ⁇ 10 1 to 3.0 ⁇ 10 3 /g" means that the temperature is measured at one or more points during the time period when the number of bacteria is in the range of 1.0 ⁇ 10 1 to 3.0 ⁇ 10 3 /g, for example, immediately after the start of fermentation, the midpoint of the fermentation period, and the end of fermentation, and the average temperature is more than 40° C. and less than 65° C.
- the specified time (fermentation time) for maintaining the fermentation temperature range can be 5 to 23 hours.
- the lower limit of the fermentation time can be 5 hours or more, preferably 6 hours or more, and more preferably 7 hours or more.
- the fermentation time if the fermentation time is shorter than the specified time, the fermentation does not proceed sufficiently, and certain components are likely to fall outside the specified range.
- the upper limit of the fermentation time is 23 hours or less, preferably 22 hours or less, and more preferably 21 hours or less. If the fermentation time is longer than the specified time, the fermentation will proceed too much, and the content of lower fatty acids in particular will increase, which is not preferable.
- the fermentation conditions are preferably such that the product temperature is between 30°C and 60°C and the fermentation time is between 5 hours and 23 hours.
- the raw material can be inoculated with Bacillus subtilis natto and fermented at a high temperature by maintaining a product temperature of 40°C to 75°C for 4 to 22 hours.
- the fermentation temperature in the case of high-temperature fermentation may be 40°C or higher and 70°C or lower.
- the lower limit can be 40°C or higher, preferably 45°C or higher, more preferably 46°C or higher, and even more preferably 47°C or higher.
- the upper limit can be 70°C or lower, more preferably 65°C or lower, even more preferably 60°C or lower, even more preferably 54°C or lower, especially preferably 53.75°C or lower, and especially more preferably 53.5°C or lower.
- the high-temperature fermentation time may be 4 hours or more and 22 hours or less.
- the lower limit is preferably maintained at 4 hours or more, preferably 5 hours or more, more preferably 8 hours or more, and even more preferably 10 hours or more.
- the upper limit is preferably maintained at 22 hours or less, preferably 20 hours or less, more preferably 18 hours or less, and even more preferably 16 hours or less.
- the average fermentation temperature is preferably more than 40° C. and less than 65° C. and the average soluble carbohydrate content is less than 2.0 mass %, which allows efficient proliferation of bacteria capable of producing koji and PGA.
- the average fermentation temperature is more than 45°C and less than 75°C, and the average soluble carbohydrate content is 2.0 mass% or more. This allows the enzyme activity derived from the koji to function efficiently, and the average soluble carbohydrate content can be further improved. This also allows the stringiness of the fermented food to be suppressed. Although the principle is unclear, it is thought that PGA production is suppressed by continuing to supply sugar during the growth of the bacteria capable of producing PGA.
- steps (II) and (I) are arbitrary; steps (II) and (I) may be performed simultaneously, step (II) may be performed before step (I), or step (II) may be performed after step (I). From the viewpoint of suppressing stringiness of the fermented food, it is preferable to perform step (II) after step (I), and it is more preferable to perform steps (II) and (I) simultaneously.
- the average soluble carbohydrate content is increased by 0.2 mass% or more by steps (I) and/or (II).
- a preferred embodiment of the present invention is natto, a fermented food made from beans and natto bacteria.
- the method for producing natto is preferably to inoculate koji and natto bacteria into beans that have been heat-treated at 80°C or higher under conditions of a moisture content of 50% or more by mass on a dry basis, and then ferment them.
- the koji may be inoculated into the beans before the natto bacteria, or the natto bacteria may be inoculated into the beans before the koji, but it is preferable to inoculate the koji and natto bacteria into the beans at the same time.
- the method for producing natto of the present invention is It is preferable that, when the number of the natto bacteria in the process is 1.0 x 10 1 to 3.0 x 10 3 /g, the average fermentation temperature is more than 40°C and less than 65°C and the average soluble carbohydrate content is less than 2.0 mass%, and, when the number of the natto bacteria in the process is more than 3.0 x 10 3 /g, the average fermentation temperature is more than 45°C and less than 75°C and the average soluble carbohydrate content is 2.0 mass% or more.
- the fermentation temperature in fermentation step 2 is, for example, from 55°C to 62°C, more preferably from 57°C to 61°C, and even more preferably from 59°C to 60.5°C.
- the fermentation time in fermentation step 2 is, for example, from 0.5 hours to 3 hours, and preferably from 1 hour to 2 hours.
- the temperature at a low level, usually between 3°C and 10°C, preferably between 3°C and 8°C, and more preferably between 3°C and 6°C, and mature the mixture for 6 hours to 3 days, preferably between 8 hours to 2 days, and more preferably for about 24 hours.
- the present invention relates to an agent for suppressing the fermented odor and grassy odor of fermented foods (the agent of the present invention), which contains at least one selected from the group consisting of methyl palmitate and ethyl palmitate.
- the properties of the agent of the present invention are not particularly limited, and examples thereof include solids (e.g., powders), semisolids, and liquids.
- the agent of the present invention can be used, for example, as a food additive, seasoning, and the like.
- food additives are used in the food manufacturing process by adding, mixing, infiltrating, or other methods.
- the other components are not particularly limited as long as they are components that can be added to food, and examples of such other components include carriers that can be added to food (e.g., excipients, binders, disintegrants, disintegration aids, lubricants, wetting agents, etc.) and additives that can be added to food.
- the form of the food additive is not particularly limited, and examples of such other forms include granules, powders, tablets, pills, and capsules (including hard and soft capsules).
- the agent of the present invention can be added to fermented foods to suppress fermentation odors and grassy odors.
- the present invention relates to a method for producing a fermented food with reduced fermentation odor and grassy odor, which comprises adjusting the methyl palmitate content in the fermented food to 0.01 ppb or more and/or adjusting the ethyl palmitate content in the fermented food to 0.01 ppb or more.
- the timing of blending methyl palmitate, ethyl palmitate, and other ingredients blended as needed is not particularly limited. Examples of such timing include during the production of the fermented food, after the production of the fermented food, and before eating.
- the origin of methyl palmitate and ethyl palmitate is not particularly limited as long as it is from an origin suitable for food, and these ingredients are derived from, for example, preparations such as flavors, food additives, seasonings, food ingredients, etc.
- After blending methyl palmitate, ethyl palmitate, and other ingredients blended as needed it is preferable to mix them as needed so that they are dispersed as uniformly as possible. For example, before eating natto, which is a fermented bean food, a liquid seasoning containing methyl palmitate and/or ethyl palmitate may be blended and mixed before eating.
- test Methods Various measurement and evaluation methods used in the following test examples are described below.
- the composition subjected to fermentation is referred to as the "pre-fermentation composition”
- the composition obtained after fermentation is referred to as the "fermented food.”
- the starch content (mass%) of the pre-fermentation composition and the fermented food was measured according to the method of the Standard Tables of Food Composition in Japan, 2015 Edition (7th Edition), using AOAC method 996.11 (AOAC, 2005).
- the starch content (mass%) of edible plants was calculated with reference to the "starch” section of the Standard Tables of Food Composition in Japan, 2015 Edition (7th Edition).
- Palmitic Acid Content The palmitic acid content in the edible plants used as raw materials was calculated with reference to the "Palmitic acid” section of the Standard Tables of Food Composition in Japan, 2015 Edition (7th Revised Edition).
- acid decomposition was performed as follows: 10 ml of nitric acid was added to 0.5 g of the sample (pre-fermentation composition), and then heat treatment was performed under the following conditions using an acid decomposition device Digiprep (manufactured by GL Sciences). (Conditions) Heat up (25 minutes) ⁇ Hold at 60°C (30 minutes) ⁇ Heat up (25 minutes) ⁇ Hold at 105°C (60 minutes).
- the acid digested solution after the above treatment was diluted with 1% nitric acid to 5,000 times the volume of the sample, and then subjected to sodium analysis using an ICP-MS 7000 (Agilent).
- the measured sodium content was converted to the salt equivalent (2.54 times) to calculate the salt content.
- the viable cell count of Bacillus subtilis natto in the samples was measured by a method in which the diluted solution obtained by diluting the sample suspension was cultured on an agar medium and the number of colonies was counted. That is, the sample was placed in a filter-equipped bag attached to a paddle-type blender "Stomacher", phosphate buffer was poured in, and the sample was shaken with the Stomacher.
- the sample was mixed in a standard agar medium (manufactured by Atect Co., Ltd.) and cultured at 37°C for 18 hours, and the number of colonies that appeared was calculated.
- a standard agar medium manufactured by Atect Co., Ltd.
- the number of colonies that appeared was significantly smaller, and the appearance of the colonies that appeared was clearly not that of bacillus such as Bacillus subtilis natto, but rather that of yeast or mold, so it was not detected (ND).
- the soluble carbohydrate content in the samples was measured as follows: Using high performance liquid chromatography in accordance with the method for measuring "available carbohydrates (glucose, fructose, galactose, sucrose, maltose, lactose, and trehalose)" in the "Analysis Manual for the 2015 Edition (7th Revised) of the Standard Tables of Food Composition in Japan,” the content was compared with that of standard monosaccharides or oligosaccharides (2-10 sugars) of known concentrations, and the total value was calculated.
- moisture content on dry basis The moisture content on a dry basis (moisture content) in the samples (pre-fermentation composition and fermented food) was measured as follows.
- DHS dynamic headspace
- GC/MS gas chromatography mass spectrometer
- the components in the sample were quantified by comparing the integration results of the peak areas of the most abundant ions in the diluted standard and sample at retention times considered to be the target components when compared with the retention times of the standard samples by analysis based on the mass spectrum pattern of a mass spectrometer.
- m/z refers to a value detected in the range of -0.3 to +0.7 at the central m/z value of each component.
- concentration of each component contained in each sample was calculated from the peak area of each component obtained, taking into account the dilution rate with the solvent.
- Preprocessing For samples used in the PGA and levan analysis of fermented foods, the samples were prepared by carrying out the deproteinization process described below, followed by a purification process using ethanol to obtain a viscous substance extract from the fermented food. 1. 10 g of the fermented food was added with 50 mL of 2.5% trichloroacetic acid (hereinafter also referred to as TCA), heated to 50° C. for 10 minutes, and stirred. 2. After removing the solid portion from the mixture, the mixture was centrifuged (12,000 rpm, 10 min) to obtain the supernatant. 3.
- TCA trichloroacetic acid
- the obtained supernatant was adjusted to pH 7.0 with sodium hydroxide and diluted two-fold with ion-exchanged water, after which an equal amount of ethanol that had been pre-cooled to -30°C was added to the neutralized liquid and stirred. 4. After standing on ice for 10 minutes, centrifugation (12,000 rpm, 10 min) was performed, the supernatant was discarded, and the mixture was dried. 5. The precipitate obtained by drying was used as a viscous substance, and the solution was dissolved in 20 mM phosphate buffer (pH 7.0) to use as a viscous substance extract.
- PGA in the fermented food was measured by measuring the absorbance of the viscous substance extract that had been pretreated as described above and the solution that had been treated in the following manner, and comparing the absorbance with that of a standard substance to perform a quantitative analysis.
- the absorbance was measured using a UV-1800 UV-visible spectrophotometer (Shimadzu Corporation). 1.
- standard solutions of 0, 25, 50, and 100 ⁇ g/mL of "poly- ⁇ -glutamic acid" average molecular weight 1,500,000 to 2,500,000
- FUJIFILM Wako Pure Chemical Corporation 20 mM phosphate buffer (pH 7.0) were prepared as PGA standard solutions. 2.
- the viscous substance extract obtained by the above-mentioned method was diluted with 20 mM phosphate buffer (pH 7.0) so that it fell within the concentration range of the calibration curve. 3.
- 20 mM phosphate buffer (pH 7.0) was added to 0.5 mL of the PGA standard solution and the analytical sample, 2.0 mL of 20 mM phosphate buffer (pH 7.0) was added, and 0.5 mL of 0.1 M cetylmethyltrimethylammonium bromide (Cetablon)/1 M NaCl solution was added and stirred. 4.
- Abs (absorbance) at a wavelength of 400 nm was measured, and the PGA concentration of the sample was calculated using a calibration curve prepared from the standard solution.
- the levan in fermented foods was measured by measuring the absorbance of the extract of the viscous substance that had been pretreated as described above and the solution that had been treated in the following manner, and quantitative analysis was performed by comparing the absorbance with that of a standard substance that used fructose as an index.
- a UV-1800 ultraviolet-visible spectrophotometer (Shimadzu Corporation) was used to measure the absorbance. 1.
- fructose standard solutions of 0, 10, 25, 50, and 100 ⁇ g/mL diluted in 20 mM phosphate buffer (pH 7.0) were prepared. 2.
- the thickening substance extract obtained above was diluted with 20 mM phosphate buffer (pH 7.0) according to the concentration so that it fell within the range of the above calibration curve. 3. 0.6 mL of the fructose standard solution and the analytical sample were placed in a test tube containing 0.3 mL of resorcinol-thiourea reagent, and 2.1 mL of 30% HCl was gently mixed. The mixture was covered with a marble or similar to prevent bumping, and incubated at 80°C for 10 minutes. 4. Abs (absorbance) was measured at a wavelength of 500 nm, and the levan concentration was calculated using a calibration curve prepared from the standard solution. The levan concentration was calculated by subtracting the water in the binding portion and multiplying the fructose calibration curve by 0.9.
- sweetness The test was carried out by four experienced panelists, who evaluated the strength of the product on the following five-level scale. 5: Strongly sweet taste. 4: Tastes sweet. 3: Slightly sweet taste. 2: Slightly sweet taste. 1: No sweetness is felt.
- Fermentation test samples A1 to A7 made from various edible plants were produced as follows. (However, among the above, sample A3 was not inoculated with Bacillus subtilis natto and was fermented only with koji. The beans and grains were used as they were without adjusting the size of the raw materials. The vegetables and potatoes were cut into pieces of 50 mm square or less using a knife or the like. The raw materials other than coconut milk were lightly washed with water and immersed in water and treated in a refrigerator for 18 hours to allow the raw materials to absorb enough water, and then the water was drained. The drained soaked raw materials were then subjected to a steaming process.
- the soaked raw materials were placed in a metal container, placed in a steaming kettle (a test steaming kettle manufactured by Harada Sangyo Co., Ltd.), heated to a temperature of 98°C, pressurized under the conditions shown in Table 1 below, and then depressurized and steamed.
- a steaming kettle a test steaming kettle manufactured by Harada Sangyo Co., Ltd.
- Table 1 a test steaming kettle manufactured by Harada Sangyo Co., Ltd.
- sample A3 only koji was mixed in using the method described below.
- koji was mixed into the steamed raw material immediately after steaming using the method described below, and natto bacteria was then inoculated.
- natto fermentation After completion of the above-mentioned procedure, the steamed raw material and coconut milk were inoculated with the following procedure.
- natto bacteria Miyagino bacteria (manufactured by Miyagino Seizosho), Bacillus subtilis K-2 strain (NITE BP-1577), and Bacillus subtilis MZ-21113 strain (NITE BP-02420) were used. These natto strains were inoculated into 10 ml/test tube of spore formation medium (YE) shown in Table 2 below, and cultured with shaking at 37° C. and 200 ⁇ g for 24 hours to obtain spore suspensions.
- YE spore formation medium
- the natto bacteria strains used were the above-mentioned natto bacteria strains, and the spore suspensions cultured in liquid media were diluted with water to about 1,000 natto bacteria per gram of steamed beans, and inoculated into the steamed raw material.
- the amount of koji described in Table 3 below was added and mixed.
- yellow koji powder koji mold: Aspergillus oryzae, manufactured by Hishiroku Co., Ltd.
- rice koji which is rice koji
- white koji powder koji mold: Aspergillus kawachii, manufactured by Hishiroku Co., Ltd.
- the mixture was placed in a polystyrene container and fermented.
- the koji was in a powder form, and its moisture content was 15% by mass or less.
- sample A3 only the koji was mixed with coconut milk without inoculation with the natto bacteria, and dispensed into a polystyrene container.
- the controlled temperature of the chamber during fermentation of the samples (hereinafter abbreviated as room temperature) is as shown in Table 3.
- the fermentation chamber used was a natto fermentation chamber (a test fermentation chamber manufactured by Harada Corporation).
- the main fermentation was carried out at room temperature of 30 to 55° C. for 8 to 72 hours. After that, the room temperature was raised to 60° C., and high-temperature treatment was carried out for 1.5 hours (hereinafter, sometimes referred to as the 60° C. step). Sample A3 was not subjected to high-temperature treatment.
- Test Example 2 Investigation of fermentation conditions Samples were produced, analyzed, and evaluated in the same manner as sample A1 in Test Example 1, except for the presence or absence of addition of koji, the presence or absence of inoculation of Bacillus subtilis natto, and the main fermentation time as shown in Table 5. The analysis results and sensory evaluation of each component are shown in Table 6. In the table, the parts that were not measured are marked with "-".
- Test Example 3 Evaluation of Stickiness Samples were produced, analyzed, and evaluated in the same manner as samples A1, A2, or A7 in Test Example 1, except that the presence or absence of koji addition, the amount of koji addition, the strain of Bacillus subtilis natto, and the main fermentation conditions were as shown in Table 7. The analysis results of each component and the sensory evaluation are shown in Table 8.
- Test Example 4 Evaluation of stringiness 2 Samples were produced, analyzed, and evaluated in the same manner as sample A1 or A7 in Test Example 1, except that no koji was added, sugar (glucose) was added at a predetermined ratio (addition concentration (mass%)) relative to 100% by mass of the composition 0, 4, 7, or 11 hours after the start of main fermentation, and the soluble carbohydrate content and viable Bacillus subtilis natto count were measured at that time. Furthermore, sample A1 in Test Example 1 was used as a positive control. The analysis results and sensory evaluation of each component are shown in Table 9.
- Test Example 5 Evaluation in a system where methyl/ethyl palmitate was added Samples were produced, analyzed, and evaluated in the same manner as samples A1 or A7 in Test Example 1 , except that the presence or absence of inoculation of Bacillus subtilis natto was as shown in Table 10, koji was not added, and methyl palmitate and/or ethyl palmitate were added after the end of fermentation so that the final concentration in the fermented food was a predetermined concentration. The analysis results and sensory evaluation of each component are shown in Tables 10 to 13. In the tables, the parts that were not evaluated are marked with "-".
- Test Example 6 Analysis of Koji ⁇ Particle size distribution measurement> The d50 after ultrasonic treatment of the yellow koji powder and the white koji powder used in the above test examples was measured as follows.
- the measurement was performed using a laser diffraction particle size distribution analyzer (Microtrac MT3300 EXII, manufactured by Microtrac Bell). Ethanol was used as the solvent during the measurement, and DMS2 (Data Management System version II, manufactured by Microtrac Bell) was used as the measurement application software.
- the measurement application software was washed by pressing the cleaning button, and then the SetZero button of the software was pressed to perform zero adjustment, and the sample was directly loaded until the appropriate concentration range was reached by sample loading.
- the ultrasonic processing button of the software was pressed, and ultrasonic processing was performed at 30 kHz, 40 W, and 180 seconds, and three degassing processes were performed, and the results of laser diffraction at a flow rate of 50% and a measurement time of 10 seconds were used as the measured values.
- the measurement conditions were set as follows: distribution display: volume; particle refractive index: 1.60; solvent refractive index: 1.36; upper measurement limit ( ⁇ m): 2000; lower measurement limit ( ⁇ m): 0.021.
- the d50 of the yellow koji powder used in the above test example after ultrasonic treatment was 484 ⁇ m
- the d50 of the white koji powder after ultrasonic treatment was 456 ⁇ m.
- the measurement was performed by heating to 90°C using the vacuum heating drying method. Specifically, an appropriate amount of sample was taken into a weighing container (W0) that had already been brought to a constant weight, weighed (W1), and placed in a vacuum electric constant temperature dryer adjusted to a specified temperature (more specifically, 90°C) at normal pressure with the mouth of the weighing container open. The door was closed, the vacuum pump was operated, and the sample was dried at the specified reduced pressure for a certain period of time.
- the vacuum pump was stopped, dry air was pumped in to return the sample to normal pressure, the weighing container was removed, the container was covered, and the sample was allowed to cool in a desiccator, after which the mass was measured.
- the drying, cooling, and weighing (W2) were repeated until a constant weight was reached, and the dry weight moisture content (mass%) was calculated using the following formula.
- the moisture content on a dry basis of the yellow koji powder used in the above test example was 2.6% by mass, and the moisture content on a dry basis of the white koji powder was 2.4% by mass.
- the ⁇ -amylase activity of yellow koji powder and white koji powder was measured using the following procedure.
- the rate of absorbance decrease C (%) during the 30-minute enzyme reaction of each measurement sample was calculated from the rate of absorbance decrease in the enzyme reaction area (absorbance A) relative to the control area (absorbance B) ( ⁇ (absorbance B - absorbance A) / absorbance B ⁇ x 100 (%)).
- the enzyme activity that reduces absorbance by 10% per 10 minutes is defined as 1 unit (U), and the enzyme activity per 1 g of dry mass of the measurement sample (U/g) was calculated from the rate of absorbance decrease C (%) when the enzyme reaction was carried out for 30 minutes using 0.25 mL of enzyme solution (sample content 0.025 g) using the following formula.
- the ⁇ -amylase activity of the yellow koji powder used in the above test example was 1140 U/g
- the ⁇ -amylase activity of the white koji powder was 1120 U/g.
- the ⁇ -amylase activity of fermented foods can be calculated from the "ratio of koji to ingredients.” Similar results were also obtained when yellow koji powder and white koji powder with ⁇ -amylase activity of 100 U/g were used.
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Abstract
La présente invention aborde le problème de la fourniture d'un aliment fermenté dans lequel des odeurs de fermentation et des odeurs brutes sont davantage supprimées. La présente invention concerne un aliment fermenté contenant un produit de fermentation d'au moins un type de plante comestible choisi dans le groupe constitué par les graines, les céréales, les haricots, les légumes, les pommes de terre, les champignons et les fruits, la teneur en palmitate de méthyle étant supérieure ou égale à 0,01 ppb et/ou la teneur en palmitate d'éthyle étant supérieure ou égale à 0,01 ppb.
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MIYUKI YAMASHITA, HIROSHI HAMADA, MISAO KOJIMA, KEIICHI YAMASHIRO: "Changes of Several Components in Saké Cake during Aging at Various Temperatures", NIPPON JOZO KYOKAI ZASSHI - JOURNAL OF THE SOCIETY OF BREWING,JAPAN, NIPPON JOZO KYOKAI, TOKYO,, JP, vol. 77, no. 11, 15 November 1982 (1982-11-15), JP , pages 825 - 830, XP009554289, ISSN: 0369-416X, DOI: 10.6013/jbrewsocjapan1915.77.825 * |
SHOICHI YAMADA: "Some Problems Concerning Synthetic Sake", NIPPON JOZO KYOKAI ZASSHI - JOURNAL OF THE SOCIETY OF BREWING,JAPAN, NIPPON JOZO KYOKAI, TOKYO,, JP, vol. 36, no. 8, 15 August 1941 (1941-08-15), JP , pages 562 - 565, XP009554290, ISSN: 0369-416X, DOI: 10.6013/jbrewsocjapan1915.36.562 * |
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