Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
  • C12G3/00—Preparation of other alcoholic beverages
  • C12G3/04—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs
  • C12G3/06—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs with flavouring ingredients
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
  • C12G3/00—Preparation of other alcoholic beverages
  • C12G3/02—Preparation of other alcoholic beverages by fermentation
  • C12G3/021—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
  • C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
  • C12H1/02—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material
  • C12H1/06—Precipitation by physical means, e.g. by irradiation, vibrations
  • C12H1/063—Separation by filtration
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
  • C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
  • C12H1/22—Ageing or ripening by storing, e.g. lagering of beer

Definitions

• the present invention relates to a method for producing a fermented malt beverage having a sufficiently reduced carbohydrate content and a fermented malt beverage produced by the method even when the proportion of malt used in the fermentation raw material is increased.
• the beer-taste beverage is an effervescent beverage having a flavor / taste and texture equivalent to or similar to beer.
• the sugar content in the beer-taste beverage that is the final product can be reduced by reducing the content of non-assimilable sugar in the fermentation liquid. it can.
• the sugar content can be reduced by increasing the use ratio of liquid sugar with a low content of non-assimilable sugar, but when the use ratio of the grain raw material is high While increasing the grain aroma and richness, the sugar content tends to be high.
• glucoamylase or pullulanase As a method for producing a beer-taste beverage having a low sugar content, for example, a method of adding glucoamylase or pullulanase during a preparation process or a fermentation process is known.
• these enzymes By the action of these enzymes, most of the starch that contributes to carbohydrates contained in the final product can be decomposed into sugars that can be assimilated by yeast.
• non-assimilable branched-chain sugars such as isomaltose are not converted into assimitable sugars by these enzymes and remain in the final product. For this reason, saccharide
• glucose is produced as an assimilating sugar to increase the degree of intrinsic fermentation, thereby containing non-assimilating sugar in the final product.
• a method for reducing the amount is known (for example, see Patent Document 1).
• the content of non-assimilable sugar in the final product could not be reduced sufficiently by simply adding transglucosidase during fermentation.
• Transglucosidase generates glucose by a hydrolysis reaction, but performs a transglycosylation reaction when the substrate concentration is high. Therefore, for example, in beer-taste beverages made from malt and processed through a fermentation process, high concentrations of isomaltoligosaccharides such as isomaltose and panose are added to the product by adding transglucosidase before heat treatment in the preparation process. As a result, the richness is enhanced (see, for example, Patent Document 2).
• the object of the present invention is to provide a fermented malt beverage having a remarkably low sugar content regardless of the proportion of malt used in the fermentation raw material, and a method for producing the fermented malt beverage.
• the present inventors have added glucoamylase in at least one of the preparation step and the fermentation step in the production of a fermented malt beverage, and by adding transglucosidase in the fermentation step, The present inventors have found that a fermented malt beverage having a significantly reduced sugar content can be produced even when the malt use ratio in the fermentation raw material is increased, and the present invention has been completed.
• the present invention has the following configuration: (1) A method for producing a fermented malt beverage, comprising using malt as a raw material, adding glucoamylase in at least one of a charging step and a fermentation step, and adding transglucosidase in the fermentation step, (2) The method for producing a fermented malt beverage according to (1) above, wherein 3 ug / g or more of transglucosidase is added to the extract concentration, (3) The method for producing a fermented malt beverage according to (1) or (2), wherein the fermentation step is performed at 5 to 15 ° C., (4) The method for producing a fermented malt beverage according to any one of (1) to (3), wherein the amount of malt used is 65 to 100% by mass in the fermentation raw material, (5) The method for producing a fermented malt beverage according to any one of (1) to (4) above, wherein the sugar in the fermented malt beverage is less than 0.5 g / 100 mL, and (6) Use of malt A fermented
• the method for producing a fermented malt beverage according to the present invention makes it possible to produce a fermented malt beverage having a significantly low carbohydrate content regardless of the malt use ratio in the fermentation raw material.
• a fermented malt beverage is a beverage produced using a malt as a raw material and undergoing a fermentation process.
• the fermented malt beverage produced by the method for producing a fermented malt beverage according to the present invention (hereinafter sometimes referred to as “the production method according to the present invention”) may be an alcoholic beverage, and the alcohol content is 1% by volume. It may be a so-called non-alcoholic beverage or low-alcoholic beverage that is less than
• liqueurs obtained by mixing malt as a raw material and a beverage produced through a fermentation process with an alcohol-containing distillate may be used.
• the alcohol-containing distillate is a solution containing an alcohol obtained by a distillation operation, and those generally classified as spirits such as spirits can be used.
• the fermented malt beverage produced by the production method according to the present invention is preferably a beer-taste beverage. Specific examples include liqueurs obtained by mixing beer, happoshu, beer or happoshu with an alcohol-containing distillate.
• the production method according to the present invention includes a preparation step and a fermentation step, using malt as a raw material, adding glucoamylase in at least one of the preparation step and the fermentation step, and adding transglucosidase in the fermentation step. It is characterized by that. More specifically, yeast can assimilate saccharides in the raw material by adding glucoamylase to a charging tank and / or fermentation tank for saccharifying the raw material and adding transglucosidase to the fermentation tank. A fermented malt beverage is produced by promoting the reaction of decomposing into fresh sugar. As a result, a non-assimilable sugar in the raw material is reduced, and a fermented malt beverage having a low carbohydrate content can be produced.
• the remaining amount of non-assimilable isomaltoligosaccharides such as isomaltose and nigerose in the final product can be greatly reduced. Since isomaltoligosaccharide is not normally assimilated by yeast, it remains as a carbohydrate component in the final product. On the other hand, in the production method according to the present invention, glucoamylase and transglucosidase are added at an appropriate timing to be decomposed into glucose and further assimilated to yeast by fermentation, resulting in the sugar remaining in the final product. Quality decreases.
• the glucoamylase used in the present invention is not particularly limited as long as it is an enzyme having a catalytic activity of cleaving the non-reducing end of starch and producing glucose, and glucoamylases derived from various organisms are used. be able to.
• any commercially available glucoamylase may be used, or these may be used in combination.
• Glucoamylase works to decompose starch in both the charging process and the fermentation process. For this reason, in the production method according to the present invention, glucoamylase may be added in at least one of the charging step and the fermentation step, and may be added in both steps.
• the addition time of glucoamylase is not particularly limited as long as the enzyme reaction by the glucoamylase added until the completion of the preparation step is sufficiently performed.
• glucoamylase may be added together with fermentation raw materials such as malt at the time of the preparation of mash, or may be added during the saccharification reaction.
• the addition time of glucoamylase is not particularly limited as long as the enzyme reaction by the glucoamylase added until the end of the fermentation process is sufficiently performed, but the fermentation start It is preferably added by the time, and more preferably added to the cold wort at the start of fermentation or before the start of fermentation.
• the transglucosidase used in the present invention is not particularly limited as long as it is an enzyme having a catalytic activity for degrading a saccharide by a hydrolysis reaction, and various organism-derived transglucosidases can be used.
• any commercially available transglucosidase may be used, and these may be used in combination.
• transglucosidase catalyzes a transglycosylation reaction or a hydrolysis reaction depends on the substrate concentration. Since the substrate concentration is high in the preparation process, the equilibrium tends to transglycosylation. For this reason, in order to decompose non-assimilable sugars such as isomaltoligosaccharides, it is necessary to add transglucosidase in the fermentation process.
• the addition time of the transglucosidase is not particularly limited as long as the enzyme reaction by the transglucosidase added by the end of the fermentation process is sufficiently performed, and may be added by the start of the fermentation. You may add to a fermentation liquid in the middle of a process. Moreover, the whole amount of transglucosidase to be used at one time may be added, or it may be added in a plurality of times.
• transglucosidase When transglucosidase is added in the early stage of fermentation where the substrate concentration is high, isomaltoligosaccharides are produced by the transglycosylation reaction, so that fermentation delay is likely to occur, resulting in a large residual amount of carbohydrate components that are not consumed in the fermentation. It is difficult to sufficiently reduce the carbohydrate content remaining in the product.
• glucoamylase since glucoamylase is used in combination, fermentation delay is less likely to occur than when transglucosidase is used alone, and the residual amount of saccharide components is significantly higher than before. Can be reduced.
• the addition amount of glucoamylase and transglucosidase is not particularly limited as long as each enzyme reaction is sufficiently performed, taking into consideration the type and titer of the enzyme used, reaction temperature, reaction time, etc. It can be determined as appropriate. For example, it is possible to reduce the amount of enzyme necessary for reducing the sugar content in the final product to a desired range by extending the fermentation time and the subsequent storage period. For example, when the fermentation / storage period is a period that is normally performed in the production of beer, happoshu, etc., transglucosidase is 3 U / g of transglucosidase with respect to the concentration of cereal-derived extract (soluble evaporation residue).
• the amount of transglucosidase added can be less than 3 U / g.
• the production method according to the present invention is for producing alcohol-fermented malt beverages such as beer and sparkling liquor, except that glucoamylase is added in at least one of the charging step and the fermentation step, and transglucosidase is added in the fermentation step.
• a general method can be adopted.
• the production method according to the present invention can be carried out in the steps of preparation, fermentation, storage, filtration, and filling.
• wort is prepared from fermentation raw materials containing malt as a preparation step. Specifically, first, malt or a crushed product thereof, fermentation raw materials other than malt as required, and raw water are added to a charging tank and mixed to prepare a miche. Miche can be prepared by a conventional method such as holding the miche at 35 to 70 ° C. for 20 to 90 minutes. Thereafter, the starch is gradually saccharified using the malt-derived enzyme or the enzyme added to the cheese, by gradually raising the temperature of the meat and holding it at a predetermined temperature for a certain period of time. After the saccharification treatment, the mixture is kept at 76 to 78 ° C. for about 10 minutes, and then the mash is filtered in a wort filtration tank to obtain transparent wort.
• malt those obtained by germinating barley or the like by a general malting process can be used.
• malt can be produced by immersing harvested barley, wheat, buckwheat, etc. in water and allowing them to germinate appropriately, followed by drying with hot air.
• the malt may be crushed by a conventional method.
• fermentation raw materials other than malt examples include starchy raw materials such as barley, wheat, corn starch, corn grits, rice and corn, and sugar raw materials such as liquid sugar and sugar.
• the liquid sugar is produced by decomposing and saccharifying starchy substances with an acid or a saccharifying enzyme, and mainly contains glucose, maltose, maltotriose and the like.
• the carbohydrate content in the final product can be significantly reduced even when the use ratio of the starch raw material to the fermentation raw material is high. For this reason, by using the production method according to the present invention, it is possible to increase the ratio of starchy raw materials, particularly malt, to fermentation raw materials without increasing the sugar content. Since the effects of the production method according to the present invention can be more fully exhibited, it is preferable to use only starchy raw materials as fermentation raw materials, and the malt use ratio relative to the fermentation raw materials is preferably 50% or more, and is 65 to 100%. More preferably.
• an enzyme agent such as a saccharifying enzyme such as ⁇ -amylase or prunalase, or a protease can be added to the mash.
• a saccharifying enzyme such as ⁇ -amylase or prunalase
• a protease can be added to the mash.
• spices, herbs, fruits and the like may be added as long as the effects according to the present invention are not impaired.
• the temperature and time during the saccharification treatment can be appropriately determined in consideration of the type of enzyme added such as glucoamylase, the amount of miche, the quality of the target fermented malt beverage, and the like. For example, it can be performed by holding at 60 to 72 ° C. for 30 to 90 minutes.
• a portion of malt, a part or all of barley, and a miche prepared by adding hot water to a charging kettle and mixing, after saccharification treatment, mixed with mash saccharified in the above-mentioned charging tank, You may obtain wort by filtering in a wort filtration tank.
• the obtained wort is boiled.
• the boiling method and its conditions can be determined as appropriate.
• a fermented malt beverage having a desired flavor can be produced by appropriately adding herbs, fragrances and the like before or during the boiling process.
• hops before or during the boiling process.
• the flavor and aroma of hops can be boiled out.
• the addition amount of hops, the addition mode (for example, adding in several steps) and the boiling conditions can be determined as appropriate.
• a precipitation tank called a whirlpool to remove hop koji or coagulated protein produced by boiling. Then, it cools to an appropriate fermentation temperature with a plate cooler.
• Cold wort may be used for the fermentation process as it is, or may be used for the fermentation process after adjusting to a desired extract concentration.
• yeast is inoculated into cold wort and transferred to a fermentation tank for fermentation.
• the yeast used for fermentation is not particularly limited, and can be appropriately selected from yeasts used for producing alcoholic beverages. Although it may be a top fermentation yeast or a bottom fermentation yeast, it is preferably a bottom fermentation yeast because it can be easily applied to large-scale brewing facilities.
• transglucosidase When transglucosidase is added before the start of fermentation, transglucosidase may be added to cold wort before yeast inoculation, or after adjusting the extract concentration, it may be added to cold wort before yeast inoculation. You may add with yeast.
• glycerol is a component remaining as a carbohydrate in fermented malt beverages.
• Glycerol is produced by yeast during fermentation and is generally contained in a beer-taste beverage of about 1000 to 2000 ppm.
• the fermentation temperature is preferably 5 to 15 ° C. from the viewpoint of the glycerol content and flavor, and is 5 to 13 ° C. More preferably.
• the obtained fermented liquor is aged in a storage tank, stored and stabilized under a low temperature condition of about 0 ° C., and then filtered as a filtration process.
• the target fermented malt beverage can be obtained by removing yeast and protein.
• the liqueur in liquor tax law can be manufactured in the process after the fermentation process by yeast, for example by mixing with spirits.
• the obtained fermented malt beverage is usually bottled by a filling process and shipped as a product.
• the content of saccharides in the wort or fermented malt beverage (mg / L) is the peak area obtained by detecting what was separated into disaccharides, trisaccharides and tetrasaccharides by high performance liquid chromatography with a mass spectrometer.
• a pump L-2100, an autosampler L-2200, a column oven L-2300 (manufactured by Hitachi High-Technologies), and a mass spectrometer API 3000 manufactured by AB SCIEX
• Detection Electrospray Ionization (ESI) positive, Multiple Reaction Monitoring (MRM) m / z 360.0 ⁇ 163.1, Column: Hypercarb (2.1 mm ⁇ 150 mm, 3 ⁇ m, flow rate: Thermo Scientific 60 ° C., column temperature: 60 ° C.) 0.2 mL / min, mobile phase A: 10 mmol / L aqueous ammonium acetate (0.1% by volume acetic acid), mobile phase B: methanol, gradient condition: 0 to 10 minutes (mobile phase B concentration: 0% by volume) ⁇ 10 25 minutes (mobile phase B concentration: 3% by volume) ⁇ 25-40 minutes (mobile phase B concentration: 13% by volume) ⁇ 40-50 minutes (mobile phase B concentration: 18% by volume) ⁇ 50-60 minutes (mobile) Phase B concentration: 40% by volume) ⁇ 60 to 70 minutes (mobile phase B concentration:
• the analysis conditions were set to detection: ESI positive, MRM m / z 522.5 ⁇ 325.1.
• the analysis conditions were set to detection: ESI positive, MRM m / z 684.5 ⁇ 325.1.
• Example 1 A beer-taste beverage was produced using a 200 L scale preparation facility. First, in a charging tank, 28 kg of malt pulverized product, 196 L of charged water, and 20 g / g glucoamylase (manufactured by Amano Enzyme, product name: Gluczyme NLP) are added to the malt pulverized product. A saccharified solution was produced. The obtained saccharified solution was filtered using a wort filtration tank, and hops were added to the obtained wort, followed by boiling. Next, the wort was transferred to a settling tank to separate and remove the precipitate, and then cooled to about 10 ° C.
• transglucosidase L transglucosidase L
• the fermented liquors of both samples are introduced into different fermenters, inoculated with beer yeast, fermented at about 10 ° C. for 7 days, and then aged in a storage tank for 8 days to beer-taste beverages (alcohol content: 3. 8% by volume) was obtained.
• the content of isomaltose, kojibiose, and nigerose was measured.
• the measurement results are shown in Table 1.
• the content of any saccharide was less than 5 mg / L, and the saccharide content was significantly reduced compared to the control sample to which no transglucosidase was added.
• the carbohydrate content of the test sample was 0.4 g / 100 mL. From the results, it is clear that the production method according to the present invention can produce a low calorie beer-taste beverage even when the malt use ratio is 100%.
• Example 2 A beer-taste beverage was prepared in the same manner as in Example 1 except that the content of 2 to 4 sugars in the fermentation liquid every other day from the start of fermentation to the end of fermentation and in the beer-taste beverage after aging was measured. Obtained.
• Table 2 shows the measurement results of the control samples
• Table 3 shows the measurement results of the test samples.
• “wort” indicates cold wort adjusted to 9.4% by mass of the extract before yeast inoculation.
• trehalose to cellobiose is a disaccharide
• meletitol to maltotriose is a trisaccharide
• maltotetraose is a tetrasaccharide.
• Example 3 Example except that transglucosidase (Amano Enzyme, product name: transglucosidase L) was added to the cold wort adjusted to 9.4% by mass of extract so as to be 0, 3, or 300 U / mL
• a beer-taste beverage was obtained in the same manner as in 1.
• the contents of isomaltose, kojibiose, and nigerose were measured.
• Table 4 shows the measurement results.
• the isomaltose content was reduced to less than 5 mg / L.
• the content of all three sugars was reduced to less than 5 mg / L.
• Example 4 A beer-taste beverage was produced using a 200 L scale preparation facility. First, 40 kg of malt pulverized product and 160 L of charged water were put into a charging tank, and a saccharified solution was produced according to a conventional method. The obtained saccharified solution was filtered using a wort filtration tank, and hops were added to the obtained wort, followed by boiling. Next, the wort was transferred to a settling tank to separate and remove the precipitate, and then cooled to about 10 ° C.
• glucoamylase manufactured by Amano Enzyme, product name: Gluczyme NLP
• transglucosidase manufactured by Amano Enzyme, product name
• the appearance extract (mass%) of each beer-taste beverage was measured. Table 5 shows the measurement results.
• Appearance extracts also include extracts other than carbohydrates such as dietary fiber and ash. However, in the present Example, since conditions other than the used enzyme are prepared, components other than carbohydrates in each beer-taste beverage are considered to be equivalent.
• a sensory test on the grain aroma and richness of the 2 samples of the test was conducted by a panel of 4 specialists.
• the evaluation is based on 2 points of commercially available happoshu with a malt use ratio of less than 25% and a saccharide content of less than 0.5 g / 100 mL.
• the case where it was hardly felt was set as 1, and the case where it felt very strong was set as 5.).
• the evaluation of the test 2 samples was 3.75 for the grain aroma and 3.75 for the richness, both of which were higher than the commercially available happoshu as a comparison target.
• the flavor of the test 2 sample had no unpleasant aroma derived from the addition of enzymes or raw materials such as auxiliary materials, and had a good flavor quality like beer. From this result, it is clear that the production method according to the present invention can produce a beer-taste beverage that has an improved malt use ratio and an excellent grain flavor and richness without increasing the sugar content.
• a beer-taste beverage was produced using a 200 L scale preparation facility. First, 40 kg of malt pulverized product and 160 L of charged water were put into a charging tank, and a saccharified solution was produced according to a conventional method. The obtained saccharified solution was filtered using a wort filtration tank, and hops were added to the obtained wort, followed by boiling. The wort was then transferred to a settling tank to separate and remove the precipitate and then cooled to about 5, 10 or 15 ° C. After adjusting these cold worts to 7.3% by mass of extract, each is introduced into different fermenters, inoculated with brewer's yeast, fermented at about 5, 10 or 15 ° C. for 7 days, and then stored for 7 days.
• the beer-taste beverage was obtained by aging in the tank.
• the glycerol concentration of each beer-taste beverage obtained was quantified using a commercially available glycerol quantification kit (manufactured by Cayman Chemical, product name: Glycerol Colorimetric Assay Kit). Furthermore, sensory evaluation of each beer-taste beverage was performed by a special panel of five people. Table 6 shows the results of quantitative determination and sensory evaluation of glycerol. During sensory evaluation, the number at the end of the comment indicates the number of panels that made the comment. It was observed that the higher the fermentation temperature, the higher the glycerol concentration in the beer-taste beverage.
• the method for producing a fermented malt beverage according to the present invention can produce a fermented malt beverage with a sufficiently reduced sugar content even when the proportion of malt used in the fermentation raw material is high.
• the said manufacturing method and the fermented malt drink manufactured by it can be utilized in the manufacture field

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• 2014
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