Coffee extract composition

Abstract

The purpose of the present invention is to provide a composition that does not require alkali treatment, is made from coffee beans or coffee grounds, and has excellent prebiotic effects.　According to the present invention, a coffee extract composition with excellent prebiotic effects can be obtained by treating coffee beans or coffee grounds with a prescribed combination of enzymes, namely, [I] a combination of (a) mannanase and an enzyme selected from the group consisting of (b) glucoamylase, (c) α-galactosidase, (d) peptidase, (e) lipase, (f) α-amylase, (g) deaminase, and (h) β-glucosidase, or [II] a combination of (b) glucoamylase and (c) α-galactosidase.

Description

Coffee extract composition

[0001] The present invention relates to coffee extract compositions. More particularly, the present invention relates to coffee extract compositions useful as prebiotics.

[0002] In recent years, sustainability has become a key focus in the food market, and "upcycling," which involves creating added-value substances from food waste, has become one of the key concepts. Furthermore, with health consciousness on the rise, prebiotics such as oligosaccharides that regulate the intestinal environment are attracting attention.

[0003] Coffee is the second most consumed beverage in the world after water, and the amount of coffee grounds discarded annually is said to be as much as 15 million tons. However, coffee grounds are known to contain large amounts of polysaccharides such as galactomannan, and can also be a source of high-value-added prebiotics.

[0004] Various techniques have been investigated for the purpose of producing lactic acid bacteria growth factors from coffee grounds by enzymatic treatment. For example, Non-Patent Document 1 discloses that coffee grounds are pretreated with sodium hydroxide, and mannooligosaccharides are extracted using endo-β-1,4-mannanase derived from Bacillus sp., and that this extract promotes the growth of lactic acid bacteria. Furthermore, Patent Document 1 discloses a method for producing polysaccharides by treating coffee beans and coffee grounds with alkali and then with cellulase.

[0005] Japanese Patent Application Laid-Open No. 2017-217466

[0006] RSC Advances, 2023, 13, 3773

[0007] To produce prebiotics from coffee beans or coffee grounds, alkaline treatment is typically required before enzymatic treatment to remove coffee bean structures that interfere with enzymatic treatment. However, waste alkali creates a significant workload and environmental burden.

[0008] Therefore, an object of the present invention is to provide a composition that does not require alkali treatment and has excellent prebiotic effects and is made from coffee beans or coffee grounds.

[0009] The present inventors have discovered that a coffee extract composition with excellent prebiotic effects can be obtained by treating coffee beans or coffee grounds with a combination of predetermined enzymes, namely, [I] a combination of (a) mannanase and an enzyme selected from the group consisting of (b) glucoamylase, (c) α-galactosidase, (d) peptidase, (e) lipase, (f) α-amylase, (g) deaminase, and (h) β-glucosidase, or [II] a combination of (b) glucoamylase and (c) α-galactosidase. That is, the present invention provides the following aspects.

[0010] Item 1. A coffee extract composition comprising an enzyme-treated coffee grounds, wherein the enzyme comprises a combination of (a) mannanase and (b) glucoamylase. Item 2. A coffee extract composition for use as a prebiotic, comprising an enzyme-treated coffee bean, wherein the enzyme comprises a combination of (a) mannanase and (b) glucoamylase. Item 3. The coffee extract composition according to Item 1 or 2, wherein the amount of (b) glucoamylase used per 1 U of (a) mannanase is 1.6 to 640 U. Item 4. Item 4. A coffee extract composition comprising an enzyme-treated product of coffee beans and / or coffee grounds, wherein the enzyme comprises a combination of (a) mannanase and an enzyme selected from the group consisting of (c) α-galactosidase, (d) peptidase, (e) lipase, (f) α-amylase, (g) deaminase, and (h) β-glucosidase, or a combination of (b) glucoamylase and (c) α-galactosidase. Item 5. The coffee extract composition according to Item 4, wherein the amount of the (c) α-galactosidase used per 1 U of the (a) mannanase is 30 to 3,000 U, the amount of the (d) peptidase used per 1 U of the (a) mannanase is 0.1 to 30 U, the amount of the (e) lipase used per 1 U of the (a) mannanase is 100 to 5,000 U, the amount of the (f) α-amylase used per 1 U of the (a) mannanase is 5 to 1,000 U, the amount of the (g) deaminase used per 1 U of the (a) mannanase is 0.1 to 30 U, the amount of the (h) β-glucosidase used per 1 U of the (a) mannanase is 0.5 to 100 U, and the amount of the (c) α-galactosidase used per 1 U of the (b) glucoamylase is 1.8 to 180 U. Item 7. A coffee extract composition according to any one of Items 2 to 4, which is used as a prebiotic. Item 8. A method for producing a coffee extract composition, comprising a step of enzymatically treating coffee grounds, wherein the enzyme comprises a combination of (a) mannanase and (b) glucoamylase.Item 8. A method for producing a coffee extract composition, comprising a step of enzymatically treating coffee beans and / or coffee grounds, wherein the enzyme comprises a combination of (a) mannanase and an enzyme selected from the group consisting of (c) α-galactosidase, (d) peptidase, (e) lipase, (f) α-amylase, (g) deaminase, and (h) β-glucosidase, or a combination of (b) glucoamylase and (c) α-galactosidase. Item 9. A food or drink comprising the coffee extract composition according to any one of Items 1 to 6. Item 10. A method for promoting probiotic growth, comprising a step of culturing probiotics using the coffee extract composition according to any one of Items 1 to 6. Item 11. The method according to Item 10, wherein the probiotics are lactic acid bacteria.

[0011] According to the present invention, a composition with excellent prebiotic effects can be obtained from coffee beans or coffee grounds without the need for alkali treatment.

[0012] 1. Coffee Extract Composition and Production Method Thereof 1-1. Raw Material (Biomass) The raw material (biomass) for the coffee extract composition of the present invention is coffee beans and / or coffee grounds.

[0013] The coffee beans may be ground or unground. The coffee beans may be roasted or unroasted. The origin of the coffee beans is not particularly limited, and examples include Robusta (Indonesia, Vietnam, Uganda) and Arabica (Brazil, Kilimanjaro, Peru, Colombia, Guatemala), and these coffee beans may be used alone or in combination of two or more.

[0014] Coffee grounds are the extraction residue remaining after roasted, ground coffee beans are used for coffee extraction. The coffee grounds are preferably obtained by using roasted ground beans for coffee extraction and then further performing post-extraction and / or washing. Post-extraction refers to further extracting the ground beans after coffee extraction with water, and can be performed, for example, by stirring in water at 40 to 100°C, preferably 50 to 80°C, more preferably 55 to 70°C, for 1 to 12 hours, preferably 2 to 8 hours, more preferably 3 to 5 hours. Washing can be performed by washing the ground beans after coffee extraction or after subsequent extraction with water (preferably unheated water).

[0015] As the raw material (biomass), one of the above-exemplified materials may be used alone, or two or more of them may be used in combination. In the present invention, a preferred example of the raw material (biomass) is coffee grounds, more preferably post-extracted coffee grounds, and even more preferably post-extracted and washed coffee grounds.

[0016] 1-2. Enzymes Enzymes for treating coffee beans and / or coffee grounds include the combination of [I] below or the combination of [II] below: [I] A combination of (a) mannanase and an enzyme selected from the group consisting of (b) glucoamylase, (c) α-galactosidase, (d) peptidase, (e) lipase, (f) α-amylase, (g) deaminase, and (h) β-glucosidase; [II] A combination of (b) glucoamylase and (c) α-galactosidase.

[0017] 1-2-1. (a) Mannanase Mannanase is an endo-1,4-β-mannanase (EC 3.2.1.78) that hydrolyzes the β-1,4-mannoside bond of β-1,4-mannan, glucomannan, and galactomannan.

[0018] Specific examples of mannanase include, but are not limited to, galactomannanase derived from the genus Aspergillus. Galactomannanase derived from the genus Aspergillus is also not particularly limited, and examples thereof include galactomannanase derived from Aspergillus niger.

[0019] These mannanases may be used alone or in combination of two or more. Among these mannanases, galactomannanase derived from Aspergillus niger is preferred.

[0020] When mannanase is used, the amount used is not particularly limited, but can be, for example, 1 to 100 U, preferably 2 to 70 U, more preferably 4 to 40 U, ​​even more preferably 6 to 20 U, even more preferably 7 to 15 U, and particularly preferably 8 to 12 U, per 1 g of the total amount of coffee beans and coffee grounds (equivalent to dry weight).

[0021] Regarding mannanase activity, locust bean gum (major component: [D-galactose:D-mannose (1:4)]) is used as the substrate, and the amount of enzyme that increases the reducing power equivalent to 1 μmol of mannose per minute is defined as 1 unit (1 U).

[0022] 1-2-2. (b) Glucoamylase Glucoamylase is an enzyme (EC 3.2.1.3) that hydrolyzes both the α-1,4-glucosidic bond and the α-1,6-glucosidic bond of starch in an exo-form from the non-reducing end.

[0023] Specific examples of glucoamylases include, but are not limited to, glucoamylases derived from the genus Aspergillus, such as the genus Rhizopus, Eudomyces, Penicillium, Nurospora, Trichoderma, and Mucor. Glucoamylases derived from the genus Rhizopus are also not particularly limited, and include glucoamylases derived from Rhizopus oryzae, Rhizopus delemer, and Rhizopus niveus.

[0024] These glucoamylases may be used alone or in combination of two or more. Among these glucoamylases, glucoamylases derived from the genus Rhizopus are preferred, and glucoamylases derived from Rhizopus oryzae are more preferred.

[0025] When glucoamylase is used, the amount used is not particularly limited, but examples include an amount of 0.16 to 64 U, 0.3 to 40 U, ​​or 0.5 to 30 U, preferably 0.9 to 25 U, 1.4 to 20 U, 2 to 17 U, 3 to 15 U, or 4 to 13 U, and more preferably 4.5 to 11 U, 5 to 9 U, or 6 to 8 U, per 1 g of the total amount of coffee beans and coffee grounds (equivalent to dry weight).

[0026] When glucoamylase is used, the ratio of the amount used to (a) mannanase is not particularly limited, but the amount per 1 U of (a) mannanase may be, for example, 1.6 to 640 U, ​​3 to 400 U, or 5 to 300 U, preferably 9 to 250 U, 14 to 200 U, 20 to 170 U, 30 to 150 U, or 40 to 130 U, more preferably 45 to 110 U, 50 to 90 U, or 60 to 80 U.

[0027] Regarding glucoamylase activity, one unit (1 U) is defined as the amount of enzyme that increases the reducing power equivalent to 1 mg of glucose per minute using potato starch as a substrate.

[0028] 1-2-3. (c) α-Galactosidase α-Galactosidase is an enzyme (EC 3.2.1.22) that hydrolyzes the non-reducing terminal α-D-galactose residue in α-D-galactosides such as galactose oligosaccharides, galactomannans, and galactolipids.

[0029] Specific examples of α-galactosidase include, but are not limited to, α-galactosidase derived from the genus Aspergillus. α-galactosidase derived from the genus Aspergillus is also not particularly limited, and examples thereof include α-galactosidase derived from Aspergillus niger and Aspergillus carneus.

[0030] These α-galactosidases may be used alone or in combination of two or more. Among these α-galactosidases, α-galactosidase derived from Aspergillus niger is preferred.

[0031] When α-galactosidase is used, the amount used is not particularly limited, but can be, for example, 3 to 300 U, preferably 20 to 240 U, ​​more preferably 35 to 180 U, even more preferably 40 to 120 U, still more preferably 45 to 80 U, and particularly preferably 50 to 70 U, per gram of the total amount of coffee beans and coffee grounds (converted into dry weight).

[0032] When α-galactosidase is used, the ratio of the amount used to (a) mannanase is not particularly limited, but the amount per 1 U of (a) mannanase may be, for example, 30 to 3000 U, preferably 200 to 2400 U, more preferably 350 to 1800 U, even more preferably 400 to 1200 U, even more preferably 450 to 800 U, and particularly preferably 500 to 700 U.

[0033] When α-galactosidase is used, the ratio of the amount used relative to (b) glucoamylase is not particularly limited, but examples of the amount used per 1 U of (b) glucoamylase include 1.8 to 180 U, preferably 12 to 140 U, ​​more preferably 20 to 100 U, even more preferably 25 to 75 U, even more preferably 27 to 50 U, and particularly preferably 30 to 45 U.

[0034] Regarding α-galactosidase activity, one unit (1 U) is the amount of enzyme that produces 1 μmol of p-nitrophenol in one minute using p-nitrophenyl-α-D-galactopyranoside as a substrate.

[0035] 1-2-4. (d) Peptidase In the present invention, the peptidase is an endopeptidase. A preferred example of the peptidase is aspartic protease (EC 3.4.23).

[0036] Specific examples of peptidases include, but are not limited to, peptidases derived from fungi such as those of the genus Rhizopus and Aspergillus; peptidases derived from actinomycetes such as those of the genus Streptomyces; and peptidases derived from bacteria such as those of the genus Bacillus, Geobacillus, Lactobacillus, and Lactococcus. Examples of peptidases derived from the genus Rhizopus include peptidases derived from Rhizopus oryzae. Examples of peptidases derived from the genus Aspergillus include, but are not limited to, peptidases derived from Aspergillus oryzae.

[0037] These peptidases may be used singly or in combination of two or more. Among these peptidases, peptidase derived from Rhizopus oryzae is preferred.

[0038] When a peptidase is used, the amount used is not particularly limited, but can be, for example, 0.01 to 3 U, preferably 0.03 to 1.5 U, more preferably 0.05 to 1 U, even more preferably 0.1 to 0.8 U, and even more preferably 0.2 to 0.6 U, per 1 g of the total amount of coffee beans and coffee grounds (equivalent to dry weight).

[0039] When peptidase is used, the ratio of the amount used to (a) mannanase is not particularly limited, but the amount per 1 U of (a) mannanase can be, for example, 0.1 to 30 U, preferably 0.3 to 15 U, more preferably 0.5 to 10 U, even more preferably 1 to 8 U, and even more preferably 2 to 6 U.

[0040] Regarding the peptidase activity, one unit (1 U) is defined as the activity that produces 1 μmol of p-nitroaniline per minute using L-leucyl-p-nitroanilide hydrochloride as a substrate.

[0041] 1-2-5. (e) Lipase Lipase is an enzyme (EC 3.1.1.3) that hydrolyzes the ester bond of lipids. Specifically, lipase acts on triacylglycerol (triglyceride) and, depending on the specificity of the enzyme, catalyzes a reaction that liberates fatty acids to produce diacylglycerol (DAG) or monoacylglycerol (MAG), or the reverse reaction.

[0042] Specific examples of lipases are not particularly limited, and include lipases derived from the genus Rhizopus, Aspergillus, Mucor, Rhizomucor, Thermomyces, Geotrichum, Penicillium, and Candida. Lipases derived from the genus Rhizopus are also not particularly limited, and include lipases derived from Rhizopus delemar, Rhizopus oryzae, Rhizopus arrhizus, Rhizopus niveus, and Rhizopus javenicus. Lipases derived from the genus Aspergillus are also not particularly limited, and include lipases derived from Aspergillus niger. Lipases derived from the genus Mucor are not particularly limited, and examples thereof include lipases derived from Mucor javanicus and Mucor miehei. Lipases derived from the genus Rhizomucor are also not particularly limited, and examples thereof include lipases derived from Rhizomucor miehei. Lipases derived from the genus Thermomyces are also not particularly limited, and examples thereof include lipases derived from Thermomyces lanuginosus.

[0043] These lipases may be used alone or in combination. Among these lipases, lipases derived from the genus Aspergillus are preferred, and lipases derived from Aspergillus niger are more preferred.

[0044] When lipase is used, there are no particular limitations on the amount used, but examples include an amount of 10 to 500 U, preferably 30 to 300 U, more preferably 50 to 250 U, even more preferably 80 to 200 U, still more preferably 100 to 150 U, and particularly preferably 110 to 130 U, per gram of the total amount of coffee beans and coffee grounds (converted into dry weight).

[0045] When lipase is used, the ratio of the amount used to (a) mannanase is not particularly limited, but the amount per 1 U of (a) mannanase can be, for example, 100 to 5000 U, preferably 300 to 3000 U, more preferably 500 to 2500 U, even more preferably 800 to 2000 U, even more preferably 1000 to 1500 U, and particularly preferably 1100 to 1300 U.

[0046] Regarding lipase activity, the amount of enzyme that increases 1 μmol of fatty acids per minute using olive oil as a substrate is defined as 1 unit (U).

[0047] 1-2-6. (f) α-Amylase α-Amylase is an enzyme (EC 3.2.1.1) that endo-hydrolyzes the α-1,4-glucosidic bond of sugars.

[0048] Specific examples of α-amylases include, but are not limited to, α-amylases derived from pulses, cereals, potatoes, filamentous fungi such as Aspergillus, actinomycetes such as Streptomyces, and bacteria such as Bacillus. α-amylases derived from the genus Aspergillus are also not particularly limited, and examples thereof include α-amylases derived from Aspergillus oryzae. α-amylases derived from the genus Bacillus are also not particularly limited, and examples thereof include α-amylases derived from Bacillus amyloliquefaciens, Bacillus flexus, Bacillus polymyxa, and Bacillus subtilis.

[0049] These α-amylases may be used alone or in combination. Among these α-amylases, α-amylases derived from filamentous fungi are preferred, those derived from the genus Aspergillus are more preferred, and those derived from Aspergillus oryzae are even more preferred.

[0050] When α-amylase is used, the amount used is not particularly limited, but can be, for example, 0.5 to 100 U, preferably 1 to 50 U, more preferably 5 to 40 U, ​​even more preferably 10 to 30 U, and even more preferably 15 to 25 U per 1 g of the total amount of coffee beans and coffee grounds (equivalent to dry weight).

[0051] When α-amylase is used, the ratio of the amount used to (a) mannanase is not particularly limited, but the amount per 1 U of (a) mannanase can be, for example, 5 to 1000 U, preferably 10 to 500 U, more preferably 50 to 400 U, even more preferably 100 to 300 U, and even more preferably 150 to 250 U.

[0052] Regarding α-amylase activity, one unit (1 U) is defined as the amount of enzyme that increases the reducing power equivalent to 1 mg of glucose per minute using potato starch as a substrate.

[0053] 1-2-7. (g) Deaminase Deaminases are enzymes (EC 3.5.4.6) that deaminate 5'-adenylic acid to produce 5'-inosinic acid.

[0054] Specific examples of deaminases include, but are not limited to, deaminases derived from filamentous fungi such as those of the genus Aspergillus, and actinomycetes such as those of the genus Streptomyces. Deaminases derived from the genus Aspergillus are also not particularly limited, and examples thereof include deaminases derived from Aspergillus melleus and Aspergillus oryzae. The deaminase derived from the genus Streptomyces is not particularly limited, and examples thereof include deaminases derived from Streptomyces aureus, Streptomyces avermitilis, Streptomyces cinnamoneus, Streptomyces griseus, Streptomyces murinus, Streptomyces thermoviolaceus, and Streptomyces violaceoruber.

[0055] These deaminases may be used alone or in combination. Among these deaminases, deaminases derived from filamentous fungi are preferred, more preferably from the genus Aspergillus, and even more preferably from Aspergillus melleus.

[0056] When deaminase is used, the amount used is not particularly limited, but can be, for example, 0.01 to 3 U, preferably 0.05 to 1.8 U, more preferably 0.1 to 1.2 U, even more preferably 0.2 to 0.8 U, and even more preferably 0.4 to 0.6 U, per 1 g of the total amount of coffee beans and coffee grounds (equivalent to dry weight).

[0057] When a deaminase is used, the ratio of the amount used to the (a) mannanase is not particularly limited, but the amount per 1 U of (a) mannanase is, for example, 0.1 to 30 U, preferably 0.5 to 18 U, more preferably 1 to 12 U, even more preferably 2 to 8 U, and even more preferably 4 to 6 U.

[0058] Regarding the deaminase activity, 5'AMP-2Na (adenylic acid disodium salt) is used as a substrate, and the amount of enzyme that reduces the absorbance OD265 by 0.001 in 1 minute is defined as 1 unit (1 U).

[0059] 1-2-8. (h) β-Glucosidase β-Glucosidase is an enzyme (EC 3.2.1.21) that catalyzes the hydrolysis of the β-glycosidic bond in sugars.

[0060] Specific examples of β-glucosidase include, but are not limited to, β-glucosidases derived from filamentous fungi such as those of the genera Aspergillus, Penicillium, and Trichoderma, actinomycetes such as those of the genus Streptomyces, bacteria such as those of the genus Bacillus, and those of the genus Cycas. β-glucosidases derived from the genus Aspergillus are also not particularly limited, and examples thereof include β-glucosidases derived from Aspergillus aculeatus, Aspergillus niger, Aspergillus oryzae, and Aspergillus pulverulentus. The β-glucosidase derived from the genus Penicillium is not particularly limited, and examples thereof include β-glucosidase derived from Penicillium decumbens and Penicillium multicolor.The β-glucosidase derived from the genus Trichoderma is also not particularly limited, and examples thereof include β-glucosidase derived from Trichoderma harzianum, Trichoderma longibrachiatum, and Trichoderma reesei. The β-glucosidase derived from the genus Streptomyces is not particularly limited, and examples thereof include β-glucosidase derived from Streptomyces avermitilis, Streptomyces griseus, and Streptomyces thermoviolaceus. The β-glucosidase derived from the genus Cycas is also not particularly limited, and examples thereof include β-glucosidase derived from Cycas revoluta Thumb.

[0061] These β-glucosidases may be used alone or in combination. Among these β-glucosidases, β-glucosidases derived from filamentous fungi are preferred, those derived from the genus Streptomyces are more preferred, and those derived from Penicillium multicolor are even more preferred.

[0062] When β-glucosidase is used, the amount used is not particularly limited, but can be, for example, 0.05 to 10 U, preferably 0.1 to 5 U, more preferably 0.3 to 3 U, even more preferably 0.5 to 2 U, and even more preferably 1 to 1.5 U per 1 g of the total amount of coffee beans and coffee grounds (equivalent to dry weight).

[0063] When β-glucosidase is used, the ratio of the amount used to (a) mannanase is not particularly limited, but the amount per 1 U of (a) mannanase can be, for example, 0.5 to 100 U, preferably 1 to 50 U, more preferably 3 to 30 U, even more preferably 5 to 20 U, and even more preferably 10 to 15 U.

[0064] Regarding β-glucosidase activity, the amount of enzyme that produces reducing sugars equivalent to 1 mg of glucose per minute using D(-)-salicin as a substrate is defined as 100 units (100 U).

[0065] 1-3. Enzyme Treatment Step In order to treat coffee beans and / or coffee grounds with a predetermined combination of enzymes, the coffee beans and / or coffee grounds described in "1-1. Raw material (biomass)" above can be contacted in water with the enzymes described in "1-2. Enzymes" above under enzymatic conditions. Specifically, the enzymatic treatment can be carried out by subjecting an enzyme reaction mixture containing coffee beans and / or coffee grounds and enzymes in water to conditions under which the enzymes can act.

[0066] The order in which each of the enzymes in a predetermined combination is allowed to act on the coffee beans and / or coffee grounds is arbitrary, and the enzymes in the combination may be allowed to act simultaneously, or at different times, or only some of the enzymes in the combination (when there are three or more enzymes) may be allowed to act simultaneously and the remaining enzymes may be allowed to act at different times as appropriate.

[0067] The conditions for the enzyme treatment (temperature, pH, time, etc.) are selected appropriately depending on the characteristics of the enzyme used, the characteristics of the coffee beans and / or coffee grounds, and / or the degree of prebiotic effect desired.

[0068] The temperature is not particularly limited and can be appropriately determined by a person skilled in the art depending on the optimal thermal properties of the enzyme used, etc., but examples include 10°C to 70°C, preferably 20°C to 60°C, more preferably 40°C to 60°C, and even more preferably 45°C to 55°C.

[0069] The pH is not particularly limited either and can be appropriately determined by those skilled in the art depending on the optimum pH of the enzyme used, the pH of the coffee beans and / or coffee grounds, etc., but examples include pH 2.0 to 11.0, preferably pH 3.0 to 10.0, more preferably pH 4.0 to 9.0, even more preferably pH 5.0 to 7.5, and even more preferably pH 5.5 to 7.0 or pH 5.5 to 6.5.

[0070] The time may be determined appropriately depending on the scale of the coffee beans and / or coffee grounds and the desired degree of prebiotic effect, but may be, for example, 15 minutes to 48 hours, preferably 30 minutes to 24 hours, and more preferably 45 minutes to 3 hours.

[0071] The enzyme treatment step can be terminated by adjusting the conditions under which the enzyme reaction mixture is subjected so as to deviate from the conditions under which the enzyme acts, and / or by inactivating the enzyme.

[0072] 1-4. Other Steps In addition to the enzyme treatment step described above in "1-3. Enzyme Treatment Step," the method for producing the coffee extract composition of the present invention can include any other steps as long as the effects of the present invention are obtained.

[0073] An example of another step is a step of subjecting coffee beans and / or coffee grounds to an alkali treatment beforehand, which are to be subjected to the enzyme treatment step. Alkali treatment is known as a pretreatment used in techniques for enzymatically treating coffee grounds. However, the coffee extract composition of the present invention does not require alkali treatment because it has an excellent prebiotic effect when treated with a predetermined combination of enzymes. Therefore, the method for producing the coffee extract composition of the present invention preferably does not include a step of subjecting coffee beans and / or coffee grounds to an alkali treatment beforehand, which are to be subjected to the enzyme treatment step.

[0074] Another example of such a step is to preliminarily treat the coffee beans and / or coffee grounds to be subjected to the enzyme treatment step with hot compressed water. Hot compressed water treatment is known as a pretreatment used in techniques for recovering glycans from coffee grounds. Conditions for the hot compressed water treatment include, for example, 0.15 to 0.25 MPa (preferably 0.18 to 0.22 MPa) and 140 to 260°C (preferably 160 to 220°C, or 180 to 210°C). However, the coffee extract composition of the present invention does not require hot compressed water treatment because it exhibits excellent prebiotic effects when treated with a predetermined combination of enzymes. Therefore, the method for producing the coffee extract composition of the present invention preferably does not include a step of preliminarily treating the coffee beans and / or coffee grounds to be subjected to the enzyme treatment step with hot compressed water.

[0075] Examples of other steps include a step of subjecting the reaction mixture after the enzyme treatment step to solid-liquid separation and recovering the liquid. The method of solid-liquid separation is not particularly limited, and examples include decantation, centrifugation, filtration, and the like. These methods may be used alone or in combination of two or more. The recovered liquid can be used as the coffee extract composition (liquid) of the present invention. The recovered liquid may be subjected to the water removal step described below, and then dissolved or diluted in water, and the resulting solution may be subjected to solid-liquid separation again, and the liquid may be recovered as the coffee extract composition (liquid) of the present invention.

[0076] An example of another step is a step of removing at least a portion of the water from the liquid obtained in the liquid recovery step. Any method can be selected as the method for removing water, such as a concentration or drying method. Drying methods include freeze-drying and spray-drying. The resulting concentrate or dried product can be used as the coffee extract composition (liquid or solid) of the present invention.

[0077] An example of another step is a step of mixing additives and / or other functional ingredients. This step may be performed at any timing. For example, the additives and / or other functional ingredients may be mixed with the liquid obtained in the step of recovering the liquid, or with the concentrate or dry product obtained in the step of removing at least a portion of the water.

[0078] Examples of additives include excipients, pH adjusters, colorants, flavoring agents, disintegrants, lubricants, stabilizers, etc. Examples of other functional components include other components useful for the growth of probiotics (excluding those contained in the enzyme-treated product), such as oligosaccharides and dietary fiber.

[0079] 1-5. Coffee extract composition The coffee extract composition of the present invention is a coffee extract composition containing coffee grounds and / or an enzyme-treated product of coffee grounds, obtained by a production method including the enzyme treatment step described above in "1-3. Enzyme treatment step" and, as necessary, the other steps described above in "1-4. Other steps."

[0080] The coffee extract composition of the present invention is useful as a prebiotic, and is therefore preferably used for prebiotic purposes. Prebiotics are foods that promote the growth of beneficial bacteria in the large intestine. In this case, the coffee extract composition can be incorporated into the culture medium used in "2. Foods and beverages" or "3. Method for promoting the growth of probiotics," which will be described later.

[0081] The state of the coffee extract composition of the present invention is not particularly limited, and it may be in the form of a liquid or a solid.

[0082] 2. Food and Drink The food and drink of the present invention contains the coffee extract composition described above in "1. Coffee extract composition and method for producing the same."

[0083] The food or drink of the present invention may consist of the above-mentioned coffee extract composition, or may further contain at least one of additives, components that enhance the palatability of the food or drink, nutritional components, and functional components.

[0084] Examples of additives that may or may not be contained or components that enhance the palatability of foods and beverages include sweeteners, flavorings, antioxidants, pH adjusters, thickeners (also used as excipients), emulsifiers, milk, cocoa, chocolate, etc. These additives may be used alone or in combination of two or more.

[0085] Examples of nutritional components or functional components that may or may not be contained include vitamins or vitamin-like substances (vitamin A, vitamin B (vitamin B1, vitamin B2, vitamin B6, vitamin B12, folic acid, niacin, pantothenic acid, biotin, etc.), vitamin C, vitamin D, vitamin E, vitamin P, vitamin K, CoQ10, etc.); polyphenols (isoflavones, equol, chlorogenic acid, etc.); probiotics (lactic acid bacteria, bifidobacteria, butyric acid bacteria, etc.); prebiotics (oligosaccharides, dietary fiber, etc.). These nutritional components or functional components may be used alone or in combination of two or more.

[0086] The properties of the food and drink of the present invention are not particularly limited, and may be any of liquid, semi-solid (gel, paste), solid (granules, fine granules, powder, tablet, capsule), etc. Furthermore, the food and drink of the present invention may be a non-emulsified preparation such as an aqueous preparation or an oil-based preparation, or an emulsified preparation such as an oil-in-water emulsion preparation or a water-in-oil emulsion preparation.

[0087] The food and drink products of the present invention can be classified into general food and drink products, functional food and drink products, and the like.

[0088] More specific examples of general foods and beverages include modified coffee or coffee drinks (e.g., unsweetened black coffee; sweetened black coffee with added sucrose, liquid sugar, sweeteners, etc.; cafe au lait-type coffee drinks in which dairy ingredients such as milk, skim milk powder, or fresh cream are added to unsweetened or sweetened coffee drinks), instant coffee, and coffee-containing foods (coffee-flavored frozen desserts or chilled confections such as jelly, pudding, ice cream, and popsicles).

[0089] Functional foods and beverages refer to foods and beverages that have a certain functionality for the living body, and examples include health functional foods such as foods for specified health uses (including conditional FOSHU [foods for specified health uses]) and foods with nutrient functions, functional food products, foods with functional claims, foods for special dietary uses, nutritional supplements, health supplements, and supplements (in various dosage forms such as tablets, coated tablets, sugar-coated tablets, capsules, and liquid preparations).

[0090] 3. Method for Promoting Probiotic Growth The method for promoting probiotic growth of the present invention comprises the step of culturing probiotics using the coffee extract composition described above in "1. Coffee extract composition and method for producing the same."

[0091] Probiotics are well known to those skilled in the art as live bacteria that exert beneficial effects on the host when ingested in appropriate amounts. Specific examples of probiotics include, but are not limited to, lactic acid bacteria, bifidobacteria, and butyric acid bacteria. These probiotics may be used alone or in combination of two or more. Among these probiotics, lactic acid bacteria are preferred.

[0092] In the culturing step, the probiotics can be cultured in a medium containing the coffee extract composition. The culture conditions are not particularly limited and can be determined appropriately based on the growth conditions of the probiotics.

[0093] The present invention will be specifically described below with reference to examples, but the present invention should not be construed as being limited to the following examples.

[0094] [1] Enzymes and coffee beans The enzymes and coffee beans shown in the table below were used.

[0095] [2] Method for measuring enzyme activity The activity of each enzyme was measured by the following method.

[0096] (a) Mannanase: 0.50 g of enzyme sample was weighed and diluted with water to an appropriate concentration to prepare the sample solution. 0.6 g of locust bean gum (for enzymes) was weighed, 100 mL of water was added, and the mixture was stirred. After heating in a 600 W microwave for 2 minutes, the mixture was stirred for 1 minute. The mixture was again heated in a 600 W microwave for 2 minutes, stirred for 1 minute, and cooled under running water. After cooling, 6 mL of 5 mol / L hydrochloric acid solution was added while stirring at room temperature, and the mixture was stirred for 15 minutes. 6 mL of 1 mol / L acetic acid / sodium acetate buffer (pH 5.0) was added, and the pH was adjusted to 5.0 with 0.5 mol / L sodium hydroxide solution. The mixture was adjusted to 300 mL with water, centrifuged at 8000 rpm for 15 minutes, and the resulting supernatant was used as the substrate solution.

[0097] Four mL of substrate solution was measured into a 50 mL Nessler tube and heated at 40°C for 10 minutes. Then, 1 mL of sample solution was added, shaken, and heated at 40°C for 10 minutes. 2 mL of Somogyi test solution was added to this solution, mixed, and the Nessler tube was loosely stoppered and heated in a water bath for 30 minutes. After cooling, 2 mL of Nelson test solution was added to this solution, mixed, and allowed to stand for 20 minutes. Water was added to make a 30 mL solution, and the mixture was centrifuged at 3000 rpm for 15 minutes. The supernatant was used as the test solution. Separately, 1 mL of sample solution was measured into a 50 mL Nessler tube, 2 mL of Somogyi test solution (I) was added, shaken, and then 4 mL of substrate solution was added and mixed. The Nessler tube was loosely stoppered and heated in a water bath for 30 minutes. The procedure was repeated as in the preparation of the test solution to prepare a control solution. The absorbance of the test solution and control solution at a wavelength of 750 nm was measured. Under these conditions, the amount of enzyme that increases the reducing power equivalent to 1 μmol of mannose per minute was defined as 1 unit (1 U), and the mannanase activity was calculated using the following formula.

[0098]

[0099] (b) Glucoamylase: Measurement was performed according to Method 4 of the 9th edition of the Official Standards of Food Additives, Glucoamylase Activity Test. 0.50 g of enzyme sample was weighed out and diluted with water to an appropriate concentration to prepare a sample solution. Potato starch was pre-dried at 105°C for 2 hours, and 1.0 g of the dried product was weighed out, mixed with 20 mL of water, and 5 mL of sodium hydroxide TS (2 mol / L) was gradually added with stirring to form a paste. The paste-like starch was heated in a water bath with stirring for 3 minutes, then 25 mL of water was added. After cooling, the mixture was neutralized with hydrochloric acid TS (2 mol / L) and hydrochloric acid TS (0.1 mol / L). 10 mL of 1 mol / L acetic acid / sodium acetate buffer (pH 4.5) was added, and water was added to a total volume of 100 mL to prepare a substrate solution.

[0100] Ten mL of substrate solution was measured and heated at 37°C for 10 minutes. 1 mL of sample solution was added and immediately shaken. After heating at 37°C for 10 minutes, 4 mL of Fehling's test solution was added and gently shaken. Heating was continued in a water bath for 15 minutes, followed by cooling to below 25°C. 2 mL of potassium iodide test solution and 2 mL of sulfuric acid (1 part by volume sulfuric acid diluted with water to 6 parts by volume) were added to prepare the test solution. Separately, a control solution was prepared using 10 mL of water instead of the substrate solution, and the procedure was repeated. The liberated iodine in the test solution and the control solution was titrated with 0.05 mol / L sodium thiosulfate solution. The endpoint was determined by adding 1-2 drops of soluble starch test solution near the titration end point, and the disappearance of the resulting blue color. Under these conditions, the amount of enzyme that increases the reducing power equivalent to 1 mg of glucose per minute was defined as 1 unit (1 U), and glucoamylase activity was calculated using the following formula:

[0101]

[0102] (c) α-Galactosidase: 0.105 g of p-nitrophenyl-α-D-galactopyranoside was weighed out and dissolved in 0.05 mol / L acetic acid / sodium acetate buffer (pH 5.5), followed by addition of 0.05 mol / L acetic acid / sodium acetate buffer (pH 5.5) to make a 50 mL solution. An appropriate amount of p-nitrophenyl-α-D-galactopyranoside solution was placed in a 50 mL beaker, placed in a thermostatic water bath at 37 ± 0.2 °C, and left for at least 15 minutes. 1 mL of sample solution was weighed into a test tube, placed in a thermostatic water bath at 37 ± 0.2 °C, and left for 5 minutes. Then, 2 mL of p-nitrophenyl-α-D-galactopyranoside solution was added and immediately shaken well. This solution was left in a thermostatic water bath at 37 ± 0.2 °C for exactly 15 minutes, after which 5 mL of borate buffer (pH 9.7) was added and shaken well. The absorbance (A1) of this solution at a wavelength of 405 nm was measured using water as a control. Separately, 1 mL of the sample dilution or sample solution was measured as a blank, and 5 mL of borate buffer (pH 9.7) was added and the mixture was shaken well. 2 mL of p-nitrophenyl-α-D-galactopyranoside solution was then added and the mixture was shaken well. The absorbance (A2) of this solution at a wavelength of 405 nm was measured using water as a control. Under these conditions, the amount of enzyme that produces 1 μmol of p-nitrophenol per minute is defined as one unit, and α-galactosidase activity was calculated using the following formula:

[0103]

[0104] (d) Peptidase: An appropriate amount of enzyme was weighed and dissolved or uniformly dispersed in pH 7.0 phosphate buffer (0.01 mol / L) to make a 50 mL solution, or this was further diluted 10-fold, 100-fold, or 1000-fold with the buffer to make a sample solution. 40 mg of L-leucyl-p-nitroanilide hydrochloride was weighed and dissolved in pH 7.0 phosphate buffer (0.01 mol / L, containing 12.5 μmol / L zinc sulfate) to make a 100 mL solution to make a substrate solution.

[0105] 2 mL of substrate solution was measured and heated at 37°C for 5 minutes, after which 0.5 mL of sample solution was added, shaken, and heated at the same temperature for 15 minutes. 2.5 mL of 0.2 mol / L hydrochloric acid was added and mixed to stop the reaction, creating the test solution. Separately, a comparison solution was prepared using a pH 7.0 phosphate buffer (0.01 mol / L) instead of the sample solution, following the same procedure as for the test solution. When measuring the absorbance at a wavelength of 405 nm for the test solution and comparison solution, the absorbance of the test solution was found to be greater than that of the comparison solution. If the test solution or comparison solution was turbid, the solution was centrifuged and the supernatant was measured. The activity required to produce 1 μmol of p-nitroaniline per minute was defined as 1 unit (1 U).

[0106]

[0107] (e) Lipase. Lipase activity can be measured using a method based on the lipase activity test method in the 9th edition of the Japanese Standards for Food Additives. 75 mL of olive oil and 225 mL of emulsion (20 g / L polyvinyl alcohol I (saponification 98.0-99.0 mol%)) were mixed and emulsified using a homogenizer to prepare a substrate solution. 1 mL of lipase solution was added to 5 mL of substrate solution and 4 mL of 0.1 mol / L phosphate buffer (pH 6.0), and the reaction was carried out at 37°C. After 20 minutes, the enzyme reaction was stopped by adding 10 mL of an ethanol-acetone mixture. Next, 10 mL of 0.05 mol / L sodium hydroxide solution and 10 mL of an ethanol-acetone mixture were added, and the mixture was titrated with 0.05 mol / L hydrochloric acid to a pH of 10. The amount of enzyme that increases the amount of fatty acids by 1 μmol per minute was defined as 1 unit (U).

[0108]

[0109] (f) Preparation of α-amylase substrate solution: Approximately 1 g of potato starch was precisely weighed out and dried at 105°C for 2 hours, and the weight loss was measured. Potato starch equivalent to 1.000 g of the dried material was accurately weighed and placed in a beaker. 20 mL of water was added. While mixing well, 5 mL of sodium hydroxide solution (2 → 25) was gradually added to form a paste. After heating in a water bath for 3 minutes while mixing, 25 mL of water was added. After cooling, the solution was accurately neutralized with 2 mol / L hydrochloric acid TS and 0.1 mol / L hydrochloric acid TS. 10 mL of one of the following buffer solutions was added, and water was added to make the solution exactly 100 mL.

[0110] Sample solution preparation method: When testing using the procedure, an appropriate amount of sample dilution solution was added to the sample to dissolve or suspend it in a sample concentration within a range in which the increase in reducing power was proportional to the sample concentration. The concentration was usually 0.4 to 0.8 u / mL. Filtered if necessary.

[0111] Measurement Procedure: 10 mL of substrate solution was accurately measured and heated at 37±0.5°C for 10 minutes. Then, 1 mL of sample solution was added and immediately shaken. This solution was then left at 37±0.5°C for exactly 10 minutes, after which 2 mL of alkaline tartrate solution for preparing Fehling's TS for starch digestion tests was added and immediately shaken. Next, 2 mL of copper solution for preparing Fehling's TS for starch digestion tests was added and gently shaken. The test tube was then sealed with an aluminum cap or stopper, heated in a boiling water bath (above 98°C) for exactly 15 minutes, and immediately cooled to below 25°C. Next, 2 mL of concentrated potassium iodide TS and 2 mL of diluted sulfuric acid (1→6) were added, and the liberated iodine was titrated with 0.05 mol / L sodium thiosulfate solution (for quantitative determination) (a mL). The endpoint of the titration was determined when 1-2 drops of soluble starch test solution was added as the titration approached the end point and the resulting blue color disappeared. Separately, 10 mL of water was accurately measured in place of the substrate solution, and after heating at 37±0.5°C for 10 minutes, 1 mL of the sample solution or water was added and mixed by shaking. Subsequent titrations were carried out in the same manner (b mL).

[0112] Activity Calculation Method: Under these conditions, the amount of enzyme that increases the reducing power equivalent to 1 mg of glucose per minute was defined as 1 unit.

[0113]

[0114] (g) Deaminase First, 1 ml of an enzyme sample solution containing AMP-deaminase was added to 3 ml of a solution prepared by mixing 0.017 M 5'AMP-2Na and 1 / 15 M phosphate buffer (pH 5.6) at a volume ratio of 1:2 to prepare a reaction solution, which was then allowed to react at 37°C for 15 minutes. After 15 minutes, 4 mL of a 2 v / v% aqueous perchloric acid solution was added to stop the reaction, and 100 μL was then measured. Water was added to the measured 100 μL to make a total of 5 mL, and the OD265 was measured. A blank was prepared by measuring the same amount except that the reaction time was set to 0 minutes. Under the above conditions, the amount of enzyme required to reduce the absorbance difference by 0.001 per minute of reaction was defined as 1 U.

[0115]

[0116] (h) β-Glucosidase: 0.50 g of D(-)-salicin was weighed out and dissolved in water to make a 50 mL solution, which was used as the substrate solution. 3 mL of pH 4.0 acetate buffer (0.1 mol / L) was weighed into a 50 mL Nessler tube, 1 mL of the substrate solution was added, and the mixture was heated at 40°C for 10 minutes. 1 mL of the sample solution was then added and immediately shaken, and the mixture was heated at 40°C for 30 minutes. 2 mL of Somogyi RT (I) was added to this solution, shaken, and the Nessler tube was loosely capped and heated in a water bath for 20 minutes. After cooling, 1 mL of Nelson RT was added to the solution, shaken well until the red precipitate of cuprous oxide was completely dissolved, and the mixture was left at room temperature for approximately 20 minutes. Water was added to make a 25 mL solution, which was used as the test solution. Separately, 3 mL of pH 4.0 acetate buffer (0.1 mol / L) was weighed into a 50 mL Nessler tube, 1 mL of substrate solution was added, and 2 mL of Somogyi test solution (I) was added. After shaking, 1 mL of sample solution was added. The Nessler tube was loosely capped and heated in a water bath for 20 minutes. This was then used as a control solution. The absorbance at 500 nm was measured for the test solution and the control solution. A calibration curve was created using 0.10, 0.15, 0.20, 0.25, and 0.30 mg / 5 mL glucose solutions. Under these conditions, the amount of enzyme required to produce reducing sugar equivalent to 1 mg of glucose per minute was defined as 100 units, and this was calculated using the following formula:

[0117]

[0118] Test Example 1 (Preparation of Dried Coffee Grounds) Coffee was extracted from ground coffee beans using an automatic coffee maker. Distilled water was added to the extracted ground coffee beans (coffee grounds) and stirred at 60°C and 100 rpm for 4 hours. The coffee grounds were separated using cheesecloth (bleached cloth), washed three times with distilled water, and then dried overnight at 60°C.

[0119] (Preparation of Coffee Ground Extract) 2.5 g of dried coffee grounds were placed in a 50 mL tube and immersed in 15 mL of distilled water at 4°C for 1 hour. The enzymes listed in Table 3 were added in the indicated amounts (the pH (25°C) of the enzyme reaction mixture containing coffee grounds, enzymes, and water was 6.0), and the mixture was stirred (200 rpm) at 50°C for 1.5 hours. After the enzymatic reaction, the mixture was incubated at 90°C for 3 minutes to terminate the enzymatic reaction. After collecting the supernatant, the coffee grounds were washed with an additional 10 mL of distilled water, and the washings were added to the supernatant. The supernatant was filtered through a 0.22 μm membrane filter and freeze-dried. The freeze-dried product was redissolved in 1 mL of distilled water and filtered through a 0.22 μm membrane filter, yielding an enzyme-treated coffee ground extract.

[0120] (Growth of Lactic Acid Bacteria) Lactobacillus gasseri (ATCC 33323) was used in the lactic acid bacteria growth experiment. A glycerol stock of this strain was subjected to static culture overnight at 37°C using MRS medium of the following composition. The cells were then recovered by centrifugation (3000 rpm, 5 minutes) and washed twice with 0.9% physiological saline. Lactobacillus gasseri was added to 2.25 mL of glucose-free MRS medium of the following composition so that the initial OD660 was 0.002. Furthermore, 0.25 mL of enzyme-treated coffee grounds extract was added, and the cells were subjected to static culture overnight at 37°C. After the culture, the OD660 value was measured. The culture was performed in duplicate.

[0121]

[0122] To eliminate the effect on the growth of lactic acid bacteria of components other than the enzymes (excipients, etc.) contained in the enzyme preparation used, a coffee grounds extract treated with a heat-inactivated enzyme was prepared by carrying out the same procedure as in "(Preparation of coffee grounds extract)" above, except that the enzymes shown in Table 3 were replaced with the enzymes that had been heat-inactivated (90°C, 10 minutes). Lactic acid bacteria were then cultured using this extract in the same manner as above, and the OD660 value was obtained.

[0123] The growth amount (ΔOD660) of the lactic acid bacteria was calculated based on the following formula.

[0124] (Results) The growth amount (ΔOD660) of lactic acid bacteria and the relative amount of ΔOD660 of each Example when the ΔOD660 of Comparative Example 1 was set to 1 are shown in the table below.

[0125]

[0126] As shown in Table 3, when coffee grounds were treated with a combination of mannanase and other specific enzymes (Examples 1 to 7), a higher prebiotic effect was observed compared to when the coffee grounds were treated with mannanase alone (Comparative Example 1).

[0127] Test Example 2 (Preparation of Dried Coffee Grounds) Dried coffee grounds were prepared in the same manner as in Test Example 1 (Preparation of Dried Coffee Grounds).

[0128] (Preparation of Coffee Grounds Extract) 2.5 g of dried coffee grounds were placed in a 50 mL tube and immersed in 14 mL of distilled water at 4°C for 1 hour. The enzymes listed in Table 4 were added in the indicated amounts, and the mixture was stirred (200 rpm) at 50°C for 1.5 hours. After the enzymatic reaction, the mixture was incubated at 90°C for 3 minutes to terminate the enzymatic reaction. After collecting the supernatant, the coffee grounds were washed with an additional 10 mL of distilled water, and the washings were added to the supernatant. This was filtered through a 0.22 μm membrane filter and freeze-dried. The freeze-dried product was redissolved in 1 mL of distilled water and filtered through a 0.22 μm membrane filter, yielding an enzyme-treated coffee grounds extract.

[0129] (Growth of lactic acid bacteria) A lactic acid bacteria growth test was carried out in the same manner as in "(Growth of lactic acid bacteria)" in Test Example 1. A coffee grounds extract treated with a heat-inactivated enzyme was prepared by the same procedure as in "(Preparation of coffee grounds extract)" above, except that the enzymes shown in Table 4 were replaced with the enzymes that had been heat-inactivated (90°C, 10 minutes) in advance, and the OD660 value of the coffee grounds extract treated with the heat-inactivated enzyme was used as the B value in calculating the growth amount (ΔOD660) of lactic acid bacteria.

[0130] (Results) The growth amount (ΔOD660) of lactic acid bacteria and the relative amount of ΔOD660 of each Example or Comparative Example when the ΔOD660 of Comparative Example 2 is set to 1 are shown in the table below.

[0131]

[0132] As shown in Table 4, compared to when coffee grounds were enzymatically treated with mannanase alone (Comparative Example 2), when coffee grounds were treated with a combination of mannanase and other specific enzymes (Examples 8 to 10, 12) and when coffee grounds were treated with a combination containing glucoamylase and α-galactosidase (Examples 11, 12), a high prebiotic effect was observed. Furthermore, considering that the prebiotic effect was not improved when coffee grounds were treated with glucoamylase or α-galactosidase (Comparative Examples 3 and 4), the improvement in the prebiotic effect of each Example was found to be significantly greater.

[0133] Test Example 3 (Preparation of Dried Coffee Grounds) Comparative Example 1 and Example 13 (without alkali treatment) were prepared in the same manner as "(Preparation of Dried Coffee Grounds)" in Test Example 1. Reference Example 1 (with alkali treatment) was prepared in the same manner as "(Preparation of Dried Coffee Grounds)" in Test Example 1, except that a 1.25 wt % aqueous sodium hydroxide solution was used instead of distilled water.

[0134] (Preparation of coffee grounds extract) For Comparative Example 1 and Example 13 (without alkali treatment), an enzyme-treated coffee grounds extract was obtained in the same manner as "(Preparation of coffee grounds extract)" in Test Example 1, except that the enzymes shown in Table 5 were used in the amounts shown. For Reference Example 1 (with alkali treatment), an enzyme-treated coffee grounds extract was obtained in the same manner as "(Preparation of coffee grounds extract)" in Test Example 1, except that instead of soaking the dried coffee grounds in distilled water at 4°C for 1 hour, the dried coffee grounds were soaked in phosphate buffer (pH 6.5) at 4°C for 4 hours.

[0135] (Growth of Lactic Acid Bacteria) A lactic acid bacteria growth test was conducted in the same manner as in "(Growth of Lactic Acid Bacteria)" in Test Example 1. A coffee grounds extract treated with a heat-inactivated enzyme was prepared by the same procedure as in "(Preparation of Coffee Grounds Extract)" above, except that instead of the enzymes shown in Table 5, the same enzymes that had been previously heat-inactivated (90°C, 10 minutes) were used, and the OD660 value of the coffee grounds extract treated with the heat-inactivated enzyme was used as the B value in calculating the growth amount (ΔOD660) of lactic acid bacteria. However, for Reference Example 1, the OD660 value of the coffee grounds extract that had not been alkali-treated but had been treated with the heat-inactivated enzyme was used as the B value in calculating the growth amount (ΔOD660) of lactic acid bacteria.

[0136] (Brix Measurement) The Brix value (Brix_A) of each enzyme-treated coffee grounds extract was measured. Furthermore, coffee grounds extracts treated with heat-inactivated enzymes were prepared by the same procedure as in "(Preparation of Coffee Grounds Extract)" above, except that instead of the enzymes shown in Table 5, the same enzymes that had been heat-inactivated (90°C, 10 minutes) were used, and the Brix value (Brix_B) of the coffee grounds extracts treated with the heat-inactivated enzymes was measured. The value obtained by subtracting Brix_B from Brix_A was used as the evaluation value of solubility. However, for Reference Example 1, the Brix value of the coffee grounds extract that was not alkali-treated and was treated with the heat-inactivated enzyme was used as Brix_B in calculating the evaluation value of solubility.

[0137] (Results) The growth amount (ΔOD660) of lactic acid bacteria, the relative amount of ΔOD660 of each Example or Comparative Example when the ΔOD660 of Comparative Example 1 is set to 1, and the solubility are shown in the table below.

[0138]

[0139] As shown in Table 5, compared to when non-alkali-treated coffee grounds were enzymatically treated with mannanase alone (Comparative Example 1), when non-alkali-treated coffee grounds were enzymatically treated with a combination of mannanase and another specific enzyme (Example 13) and when alkali-treated coffee grounds were enzymatically treated with mannanase alone (Reference Example 1), the prebiotic effect was improved and solubility was also improved. Although the solubility improvement effect when non-alkali-treated coffee grounds were treated with a combination of mannanase and another specific enzyme (Example 13) was significantly lower than the solubility improvement effect when alkali-treated coffee grounds were enzymatically treated with mannanase alone (Reference Example 1), the prebiotic effect was equivalent to that of Reference Example 1. In other words, the improvement in prebiotic effect achieved by treating coffee grounds with a combination of mannanase and another specific enzyme is not simply due to improved solubility, but is a unique effect achieved by treating coffee grounds with a specific enzyme combination.

Claims

1. A coffee extract composition comprising enzyme-treated coffee grounds, wherein the enzyme comprises a combination of (a) mannanase and (b) glucoamylase.

2. A coffee extract composition for use as a prebiotic, comprising enzyme-treated coffee beans, wherein the enzyme comprises a combination of (a) mannanase and (b) glucoamylase.

3. A coffee extract composition according to claim 1 or 2, wherein the amount of (b) glucoamylase used per 1 U of (a) mannanase is 1.6 to 640 U.

4. A coffee extract composition comprising enzyme-treated coffee beans and / or coffee grounds, wherein the enzyme comprises a combination of (a) mannanase and an enzyme selected from the group consisting of (c) α-galactosidase, (d) peptidase, (e) lipase, (f) α-amylase, (g) deaminase, and (h) β-glucosidase, or a combination of (b) glucoamylase and (c) α-galactosidase.

5. The coffee extract composition according to claim 4, wherein the amount of (c) α-galactosidase used per 1 U of (a) mannanase is 30 to 3,000 U, the amount of (d) peptidase used per 1 U of (a) mannanase is 0.1 to 30 U, the amount of (e) lipase used per 1 U of (a) mannanase is 100 to 5,000 U, the amount of (f) α-amylase used per 1 U of (a) mannanase is 5 to 1,000 U, the amount of (g) deaminase used per 1 U of (a) mannanase is 0.1 to 30 U, the amount of (h) β-glucosidase used per 1 U of (a) mannanase is 0.5 to 100 U, and the amount of (c) α-galactosidase used per 1 U of (b) glucoamylase is 1.8 to 180 U.

6. A coffee extract composition according to claim 2 or 4, used as a prebiotic.

7. A method for producing a coffee extract composition, comprising the step of enzymatically treating coffee grounds, wherein the enzyme comprises a combination of (a) mannanase and (b) glucoamylase.

8. A method for producing a coffee extract composition, comprising a step of enzymatically treating coffee beans and / or coffee grounds, wherein the enzyme comprises a combination of (a) mannanase and an enzyme selected from the group consisting of (c) α-galactosidase, (d) peptidase, (e) lipase, (f) α-amylase, (g) deaminase, and (h) β-glucosidase, or a combination of (b) glucoamylase and (c) α-galactosidase.

9. A food or beverage comprising the coffee extract composition according to any one of claims 1, 2, and 4.

10. A method for promoting the growth of probiotics, comprising culturing probiotics using the coffee extract composition of any one of claims 1, 2 and 4.

11. The method of claim 10, wherein the probiotic is a lactic acid bacterium.

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