WO2010032510A1 - Novel branched glucan, method for producing the same, and use of the same - Google Patents

Novel branched glucan, method for producing the same, and use of the same Download PDF

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WO2010032510A1
WO2010032510A1 PCT/JP2009/057609 JP2009057609W WO2010032510A1 WO 2010032510 A1 WO2010032510 A1 WO 2010032510A1 JP 2009057609 W JP2009057609 W JP 2009057609W WO 2010032510 A1 WO2010032510 A1 WO 2010032510A1
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branched
sugar
megalosaccharide
glucan
food
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French (fr)
Japanese (ja)
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亘 佐分利
由香里 上村
祐一郎 大畑
山本　健
正保 高田
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日本食品化工株式会社
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    • 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
    • A23L2/52—Adding ingredients
    • A23L2/66—Proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/156—Flavoured milk preparations ; Addition of fruits, vegetables, sugars, sugar alcohols or sweeteners
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G9/00—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor
    • A23G9/32—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds
    • A23G9/34—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds characterised by carbohydrates used, e.g. polysaccharides
    • 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
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/50—Poultry products, e.g. poultry sausages
    • A23L13/55—Treatment of original pieces or parts
    • A23L13/57—Coating with a layer or stuffing
    • 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
    • A23L2/02—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation containing fruit or vegetable juices
    • 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
    • A23L2/52—Adding ingredients
    • 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
    • A23L2/52—Adding ingredients
    • A23L2/54—Mixing with gases
    • 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
    • A23L2/52—Adding ingredients
    • A23L2/60—Sweeteners
    • 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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
    • A23L29/35—Degradation products of starch, e.g. hydrolysates, dextrins; Enzymatically modified starches
    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
    • 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
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin

Definitions

  • the present invention relates to a glucan having a branched structure at least at a non-reducing end and a method for producing the glucan.
  • the present invention also relates to uses of the branched glucan and foods and pharmaceuticals containing the same.
  • Starch is a natural polysaccharide composed of glucose as the only saccharide, and a branched structure is formed by binding a number of similar glucan chains to the main chain consisting of ⁇ -1,4-linkages through ⁇ -1,6-linkages. It is composed of amylopectin having a tufted structure and linear amylose having no branched structure. Starch is generally insoluble in water, but becomes water-soluble by denaturation by heat or hydrolysis to some extent by acid or enzyme to lower the molecular weight. Thereafter, by proceeding with decomposition, dextrin, maltooligosaccharide and glucose are obtained, and these are generally called starch sugar. The degree of starch degradation is expressed in terms of glucose equivalent (DE). Dextrins are obtained by relatively mildly degrading starch, but those with a DE of 10 or less are called dextrins and those with 10-20 are called maltodextrins.
  • DE glucose equivalent
  • the starch degradation products with different DEs have different physical properties depending on the DE. That is, the higher the degree of degradation, the higher the degree of sweetness and the maximum for glucose with the highest DE.
  • the lower the DE the stronger the aging property, which is the original property of starch as a raw material, and in an aqueous solution, it aggregates due to aging and produces a white precipitate. Further, this aging property becomes more prominent as the branched structure is smaller, and amylose not having a branched structure is significantly aged and cannot be stored in an aqueous solution state. Also, the viscosity of the solution strongly depends on DE.
  • starch when starch is used as a raw material for foods and drinks, it can be said that it is impractical to use a liquid product from the viewpoints of aging, viscosity, and storage stability. For this reason, starch and dextrin are generally distributed in powder form.
  • starch When starch is used in foods and drinks, the aging properties of starch and dextrin often affect the quality. That is, white turbidity due to insolubilization due to aging in beverages, and deterioration of food texture in foods. For this reason, starch and dextrin excellent in aging resistance have been demanded, and various studies have been made. In order to produce starch sugar having excellent aging resistance and excellent storage stability, starch is decomposed to such an extent that it does not exhibit aging property, that is, the degree of polymerization is 10 or less, or the starch is treated so as to have a dense branched structure. There was a need.
  • ⁇ -limit dextrin obtained by degrading starch to the limit with ⁇ -amylase and then removing the resulting maltose has many short-chain branched structures, and has excellent aging resistance even at low DE. Have.
  • ⁇ -limit dextrin it was necessary to remove maltose by fractionation.
  • a water-soluble macrocyclic glucan is formed by allowing a branching enzyme (branching enzyme), 4- ⁇ -glucanotransferase or cyclodextrin-forming enzyme to synthesize starch branch chains to act on the starch liquefaction solution.
  • Patent Document 3 Japanese Patent Laid-Open No. 8-134104 (Patent Document 3)). Furthermore, a branching starch having a branching structure with a glucose polymerization degree of 6 or 7 is formed using glucose-1-phosphate and maltooligosaccharide as substrates using a branching enzyme (SBE-II) derived from barley and phosphorylase. A method of causing this is proposed (Japanese Patent Laid-Open No. 2002-78497 (Patent Document 4)).
  • SBE-II branching enzyme
  • polyphenols are plant components having a plurality of phenolic hydroxyl groups in the molecule, and catechins, anthocyanins, chlorogenic acids and the like are known as typical polyphenols.
  • Polyphenols have been reported to have many physiological effects beneficial to health promotion.
  • catechin has an antioxidant action, an antibacterial action, an action to improve lipid metabolism, an action to suppress an increase in blood pressure, and an action to suppress an increase in blood glucose level. , Deodorant action, antiallergic action, etc. Utilizing such physiological activity, green tea beverages and near water containing catechins at high concentrations are widely sold as health beverages.
  • polyphenols exhibit strong bitterness and astringency. That is, due to the strong bitterness and astringency of polyphenols, foods and drinks containing polyphenols at a high concentration are difficult to eat and drink due to the bitterness and astringency. For this reason, although polyphenols have various beneficial physiological functions, it is a problem that it is difficult to ingest high concentrations.
  • ⁇ -cyclodextrin has an ADI value (daily acceptable intake) of 5 mg / kg / day, and there is a problem that the amount of use is limited. There was a problem that even the taste was reduced.
  • Patent Document 6 discloses a method for suppressing the bitterness and / or astringency of catechins by using a flavor regulator containing a 3-4 sugar having a branched structure as an active ingredient. .
  • a flavor regulator containing a 3-4 sugar having a branched structure as an active ingredient.
  • the lower the sugar the stronger the sweetness, and the higher the sugar content of 3-4 sugars, the more sweetness is unavoidable, which limits the use of low-sweet foods such as green tea beverages. .
  • JP-A-2008-61593 discloses a method for reducing the bitterness and astringency of polyphenols by blending maltohexaose and / or maltoheptaose in a range of 1.5 to 6% by mass. Is disclosed. However, the maltohexaose and maltoheptaose-containing syrups used in this document also contain low-molecular sugars and are easy to feel sweetness, so the amount used is limited in low-sweet drinks such as green tea drinks for the same reason as above. There was a problem of being.
  • the inventors of the present invention when an enzyme having a sugar transfer action is allowed to act on a starch liquefaction solution together with a cyclodextrin-forming enzyme, hardly produces cyclodextrin and has a degree of polymerization of about 11 to 35 having a branched structure at the non-reducing end.
  • glucan can be produced.
  • the present inventors have also shown that a glucan having a branched structure at the non-reducing end and having a polymerization degree of about 11 to 35 has extremely high aging resistance as compared with a linear maltodextrin, and improves flavor and texture. It was found that it is extremely effective.
  • the present invention is based on these findings.
  • a glucan having a degree of polymerization of 11 to 35 and having a branched structure at least at the non-reducing end, or a reduced product thereof (hereinafter referred to as “megalosaccharide according to the present invention”).
  • the glucan or reduced product thereof according to any one of (1) to (3) which comprises a step of allowing a cyclodextrin-forming enzyme and an enzyme having a glycosyltransferase action to act on a starch raw material.
  • (6) The production method according to (5), wherein a debranching enzyme is further acted on in addition to the cyclodextrin-forming enzyme and the enzyme having a glycosyltransferase effect.
  • cyclodextrins synthase Paenibacillus sp (Paenibacillus sp.), Bacillus coagulans (Bacillus coagulans), Bacillus stearothermophilus (Bacillus stearothermophilus), or Bacillus Mazeransu (Bacillus Macelans) is derived from, (5) Or the manufacturing method as described in (6).
  • the debranching enzyme is selected from the group consisting of isoamylase, pullulanase, and combinations thereof.
  • the debranching enzyme is selected from the group consisting of an isoamylase derived from Myroides odoratus, an isoamylase derived from Pseudomonas amyloderamosa, a Klebsiella pneumoniae- derived pullulanase, and a combination thereof.
  • a food additive or a pharmaceutical additive comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
  • the flavor improvement is a reduction in bitterness and / or astringency, a reduction in sourness, a reduction in taste and / or an unpleasant taste, an enhanced richness, a reduction in unpleasant odor, or a reduction in alcoholic feeling.
  • a texture improving agent comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
  • a homogeneous ice freezing agent comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
  • a shine and / or gloss improver for food comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
  • a method for improving flavor comprising adding the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4) to food. Or a method for producing a food with improved flavor.
  • a method for reducing bitterness and / or astringency, or a method for producing a food with reduced bitterness and / or astringency comprising: (22) The method for reducing bitterness and / or astringency according to (21) or the food having reduced bitterness and / or astringency, wherein the food having a bitterness and / or astringency is a polyphenol-containing food. Production method.
  • Polyphenol-containing foods are tea-based beverages, fruit beverages, carbonated beverages, vegetable beverages, sports beverages, dairy beverages, alcoholic beverages, other beverages, ice creams, jellies, mousses, sweets, gums, fillings,
  • An unpleasant taste comprising adding the glucan according to any one of (1) to (3) or a reduced product thereof or the liquid sugar or powdered sugar according to (4) to a pharmaceutical product Masking method or a method for producing a pharmaceutical product with an unpleasant taste masked.
  • Improvement of texture comprising adding the glucan or reduced product thereof according to any one of (1) to (3) or the liquid sugar or powdered sugar according to (4) to food.
  • Glucan according to any one of (1) to (3) or a reduced product thereof or liquid sugar or powdered sugar according to (4) is added to water in which the additive is uniformly dissolved or dispersed.
  • Irradiation of food comprising adding the glucan according to any one of (1) to (3) or a reduced product thereof or the liquid sugar or powdered sugar according to (4) to the food, and A method for improving the gloss or / or a method for producing food with improved shine and / or gloss.
  • a food comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
  • a polyphenol-containing food comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
  • the glucan according to any one of (1) to (3) or the reduced product thereof is contained in an amount of 1.6 to 50% by mass with respect to 0.13% by mass of the polyphenols. Foods containing polyphenols.
  • Polyphenol-containing foods are tea-based beverages, fruit beverages, carbonated beverages, vegetable beverages, sports beverages, dairy beverages, alcoholic beverages, other beverages, ice creams, jellies, mousses, sweets, gums, fillings,
  • a pharmaceutical comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
  • a milk protein-containing beverage comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
  • the branched megalosaccharide according to the present invention has excellent aging resistance, as well as excellent storage stability and operability.
  • the branched megalosaccharide according to the present invention also has a taste improving action such as masking unpleasant taste.
  • the branched megalosaccharide according to the present invention further has an action of improving and promoting ice uniformity when added to water containing a mixed component such as saccharide.
  • the branched megalosaccharide according to the present invention also has an effect of improving the shine and gloss of food.
  • the branched megalosaccharide according to the present invention has a low sweetness and does not affect the original flavor of food even when added to food.
  • the branched megalosaccharide according to the present invention can also prevent milk protein aggregation and precipitation and allow milk protein to exist stably. Therefore, the branched megalosaccharide and its reduced product according to the present invention, and liquid sugar and powdered sugar containing the same can be widely used as food additives and pharmaceutical additives.
  • the branched megalosaccharide according to the present invention can be produced simply and with high yield. Therefore, the production method according to the present invention opens up the industrial production of branched megalosaccharide according to the present invention.
  • the branched megalosaccharide according to the present invention is a glucan having a polymerization degree of 11 to 35 composed of a linear glucan and a branched structure, and a branched structure is introduced at least at the non-reducing end of the linear glucan. It is a glucan.
  • the “linear glucan” means a linear glucan composed of glucose molecules bonded by a single glucoside bond.
  • branched megalosaccharide according to the present invention include a structure comprising a linear glucan composed of ⁇ -1,4-glucoside bonds and a branched structure introduced at least at the non-reducing end of the linear glucan. And branched megalosaccharide having a polymerization degree of 11 to 35. Further specific examples of the branched megalosaccharide according to the present invention include a linear glucan constituted by ⁇ -1,4-glucoside bonds and a branched structure introduced only at the non-reducing end of the linear glucan. And a branched megalosaccharide having a polymerization degree of 11 to 35.
  • the “branched structure” means a glucan residue composed of one or more glucose residues bonded to a linear glucan by a glucoside bond other than an ⁇ -1,4-glucoside bond.
  • glucoside bonds other than ⁇ -1,4-glucoside bonds include ⁇ -1,6-glucoside bonds, ⁇ -1,3-glucoside bonds, and ⁇ -1,2-glucoside bonds.
  • the branch structure introduced into the non-reducing end can be changed by selecting an enzyme having a glycosyltransferase effect used in the production method according to the present invention.
  • the number of glucose residues constituting the branched glucan residue is not particularly limited as long as the degree of polymerization of the branched megalosaccharide according to the present invention is satisfied, but preferably 1 to several, more preferably 1 to 6, 1 There can be ⁇ 4, 1-3, or 1-2.
  • the branched structure is a glucan residue composed of disaccharide units
  • examples of the branched structure include cordobiose, nigerose, maltose, isomaltose structure, and more specifically, the non-reducing end of a linear glucan.
  • cordobiose, nigerose, maltose, or isomaltose is bonded in a bonding mode other than ⁇ -1,4-bond.
  • the branched structure may be a glucan composed only of a single glucoside bond such as isomaltotriose or nigerotriose, or a plurality of glucoside bonds such as panose. More specifically, a single glucoside bond such as isomaltotriose or nigerotriose is used at the non-reducing end of a linear glucan with a binding mode other than ⁇ -1,4-linkage. And a structure in which a glucan composed of a plurality of glucoside bonds such as panose is bound.
  • the “reducing terminal” means a sugar residue that exhibits reducing properties.
  • the “non-reducing terminal” means a sugar residue that does not exhibit reducing properties, that is, a terminal sugar residue other than the “reducing terminal”.
  • the “degree of polymerization” refers to the number of glucose residues constituting a glucan and includes not only the number of glucose residues constituting a linear glucan but also the number of glucose residues constituting a branched structure. .
  • the degree of polymerization of the branched saccharide can be measured by high performance liquid chromatography (HPLC) method.
  • the degree of polymerization of megalosaccharide according to the present invention is 11 to 35, but the degree of polymerization may be in the range of 15 to 35.
  • a glucan having a degree of polymerization of 11 to 35 is sometimes simply referred to as “megalosugar”.
  • reduced product refers to a product in which the aldehyde group of the glucosyl group at the reducing end of the sugar is reduced to form a hydroxyl group.
  • Methods for obtaining a reduced product of sugar are well known to those skilled in the art.
  • Examples of usable reduction methods include a method using a hydride reducing agent, a method using a metal in a protic solvent, an electrolytic reduction method, and a catalytic hydrogenation reaction. Methods and the like.
  • the method using a hydride reducing agent is convenient and convenient without requiring a special apparatus, while on the other hand, when carried out on an industrial scale, A method using a catalytic hydrogenation reaction is preferred because it is economical and has few by-products.
  • the catalytic hydrogenation reaction is a reaction in which hydrogen is added to the double bond portion of an unsaturated organic compound in the presence of a catalyst, and is generally also referred to as a hydrogenation reaction.
  • the method for producing the reduced product according to the present invention will be specifically described.
  • the branched megalosaccharide according to the present invention is dissolved in water, an appropriate amount of Raney nickel catalyst is added thereto, hydrogen gas is added, and reduction is performed under high temperature conditions. Next, decolorization / deionization treatment is performed to obtain a branched megalosaccharide reducing sugar composition.
  • the catalyst that can be used in the catalytic hydrogenation reaction is not particularly limited as long as it is a known hydrogenation catalyst.
  • nickel-supported on various supports such as Raney nickel, reduced nickel, diatomaceous earth, alumina, pumice, silica gel, and acid clay.
  • Nickel catalysts such as supported catalysts; cobalt catalysts such as Raney cobalt, reduced cobalt, cobalt-supported catalysts; copper catalysts such as Raney copper, reduced copper, copper-supported catalysts; palladium black, palladium oxide, colloidal palladium, palladium-carbon, Palladium catalysts such as palladium-barium sulfate, palladium-magnesium oxide, palladium-alumina; platinum catalysts such as platinum-support catalysts such as platinum black, colloidal platinum, platinum oxide, platinum sulfide, platinum-carbon; colloidal rhodium, rhodium-carbon Rhodium catalysts such as rhodium oxide; Platinum group catalysts such as um catalyst; -; copper chromium oxide catalyst; molybdenum trioxide catalyst; vanadium oxide catalyst; tungsten oxide catalyst; dinitrogen rhenium, rhenium rhenium catalyst such as carbon such as silver catalyst.
  • Raney nickel such as Raney nickel
  • the hydrogen pressure is usually in the range of 10 to 250 kg / cm 2 , preferably 50 to 200 kg / cm 2 .
  • the reaction temperature varies depending on the amount of catalyst and the type of solvent, but is usually preferably in the range of 80 to 200 ° C, more preferably 90 to 160 ° C.
  • branched megalosaccharide according to the present invention is intended to include a reduced product.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can be produced by allowing a cyclodextrin-forming enzyme and an enzyme having a sugar transfer action to act on a starch raw material.
  • the starch raw material it is preferable to cause the starch raw material to act together with a cyclodextrin-producing enzyme and an enzyme having a sugar transfer action.
  • ⁇ -glucosidase acts on the non-reducing end of dextrin contained in starch raw material, or on the non-reducing end of dextrin that has been reduced in molecular weight in any of hydrolysis, coupling, or disproportionation of cyclodextrin-forming enzyme. Then, the ⁇ -1,4-bond is cleaved, and the glucosyl group is linked to another or the same non-reducing terminal glucosyl group by ⁇ -1,6-bond, ⁇ -1,2-bond, or ⁇ -1, 3- Add with a bond.
  • the cyclodextrin-producing reaction by the cyclodextrin-producing enzyme occurs only in the early stage of the reaction, and the cyclodextrin produced by the coupling reaction of the cyclodextrin-producing enzyme is opened and used as a substrate for the transglycosylation reaction by ⁇ -glucosidase. Supplied.
  • cyclodextrin produced in the early stage of the reaction is almost completely decomposed in the later stage of the reaction, and almost no cyclodextrin remains in the later stage of the reaction.
  • a debranching enzyme coexists in the reaction solution, the starch branch chain is cleaved and linear dextrin is supplied. Since linear dextrin also functions as an acceptor molecule in the coupling reaction, it is considered that the reaction can proceed efficiently.
  • the enzymatic reaction for producing branched megalosaccharide from starch raw material can be carried out at a temperature at which the enzymatic reaction proceeds, and can usually be carried out in a temperature range up to around 60 ° C.
  • the preferred reaction temperature is 30-55 ° C.
  • the enzymatic reaction for producing branched megalosaccharide from the starch raw material can be carried out at a pH at which the enzymatic reaction proceeds, and can usually be carried out in the range of pH 5-9.
  • a suitable reaction pH is in the range of pH 5.5-7.
  • the amount of enzyme used and the reaction time are closely related, and the reaction time can be appropriately adjusted according to the progress of the target enzyme reaction.
  • a starch liquefied liquid can be used as a starch raw material.
  • the starch concentration of the starch liquefaction solution used as a raw material can be set to 10 to 45% by mass from the viewpoint of the efficiency of enzyme reaction, the solubility of starch, and the like.
  • the reaction can proceed well even if the substrate concentration exceeds 45% by mass.
  • the DE of the starch liquefaction liquid is usually preferably in the range of DE 2 to 25, more preferably in the range of DE 3 to 10.
  • starch gelatinized material may be used as a starch raw material.
  • the amount of cyclodextrin-producing enzyme used in the enzyme reaction in the production method according to the present invention can be 0.2 to 10 units per gram of substrate (solid) from the viewpoint of reaction efficiency and production cost.
  • 1 unit of cyclodextrin-producing enzyme means the amount of enzyme necessary to produce 1 mg of ⁇ -cyclodextrin per minute under the conditions of the method for measuring the activity of cyclodextrin-producing enzyme described later.
  • the amount of ⁇ -glucosidase added is 0.01 to 30 units per gram of substrate (solid) from the viewpoint of reaction efficiency and production cost. can do.
  • one unit of ⁇ -glucosidase refers to the amount of enzyme required to hydrolyze 1 ⁇ mol of maltose per minute under the conditions of the ⁇ -glucosidase activity measurement method described later.
  • a debranching enzyme can be further acted in addition to the cyclodextrin-forming enzyme and the enzyme having a sugar transfer action.
  • a debranching enzyme is allowed to act, since the residual amount of dextrin components exceeding the degree of polymerization of 35 remaining after the branching megalosaccharide production reaction is small and the amount of branched megalosaccharide production is increased, the branched megalosaccharide is efficiently produced.
  • the debranching enzyme is preferably allowed to act on the starch raw material together with the cyclodextrin-forming enzyme and the enzyme having a sugar transfer action.
  • the amount of isoamylase added can be 10 to 1000 units per gram of substrate (solid) from the viewpoint of reaction efficiency and production cost.
  • the amount of pullulanase added may be 0.001 to 0.1% by mass per substrate (solid) from the viewpoint of reactivity and production cost. it can.
  • 1 unit of isoamylase is an enzyme titer that increases the absorbance at 610 nm by 0.01 under the conditions of the method for measuring the activity of isoamylase described later.
  • addition amount of the debranching enzyme other than isoamylase and pullulanase those skilled in the art can determine the addition amount according to the addition amount of isoamylase or pullulanase.
  • the origin of the starch used as the raw material for the production method according to the present invention is not particularly limited.
  • Subsurface starches such as starch starch and waste starch can be used.
  • amylose, amylopectin, starch partial decomposition product, etc. obtained from starch can be used as a raw material.
  • These starches can be liquefied or gelatinized and used in the production method according to the present invention.
  • a starch liquefied liquid obtained by allowing a liquefying enzyme such as amylase to act on starch can be used as a starch raw material in the production method according to the present invention.
  • Methods for liquefying starch and gelatinizing are well known to those skilled in the art, and any method can be used.
  • the enzyme used in the production method according to the present invention may be a purified enzyme or a crude enzyme as long as a branched megalosaccharide can be prepared, and it may be a free enzyme or an immobilized enzyme. Also good.
  • the reaction format may be any of batch, semi-continuous and continuous.
  • a known method such as a carrier bonding method (for example, a covalent bonding method, an ionic bonding method, or a physical adsorption method), a crosslinking method, or a comprehensive method (lattice type or microcapsule type) should be used. Can do.
  • cyclodextrin-producing enzyme used in the production method according to the present invention, a commercially available product or a product isolated from a microorganism may be used.
  • the microorganism as an isolation source may be a recombinant microorganism having the ability to produce a cyclodextrin-producing enzyme, or a mutant strain obtained by mutating a naturally-occurring microorganism, in addition to a naturally-occurring microorganism.
  • Microbial origin of "cyclodextrin forming enzyme” is not particularly limited, for example, Paenibacillus sp (Paenibacillus sp.), Bacillus coagulans (Bacillus coagulans), Bacillus stearothermophilus (Bacillus stearothermophilus), and Bacillus Mazeransu (Bacillus macelans) from Can be used.
  • Examples of the “enzyme having glycosyltransferase effect” used in the production method according to the present invention include ⁇ -glucosidase, 6- ⁇ -glucosyltransferase, dextrin dextranase, and cyclic maltosyl maltose producing enzyme.
  • ⁇ -glucosidase a commercially available product or a product isolated from a microorganism may be used.
  • the microorganism as an isolation source may be a recombinant microorganism having the ability to produce an ⁇ -glucosidase-producing enzyme, or a mutant strain obtained by mutating a naturally-occurring microorganism, in addition to a naturally-occurring microorganism.
  • the microbial origin of ⁇ -glucosidase is not particularly limited. For example, those derived from Aspergillus niger and Acremonium sp. can be used.
  • Examples of the “branching enzyme” used in the production method according to the present invention include isoamylase and pullulanase, and it is preferable to use an isoamylase that can easily cleave a long branched chain.
  • the isoamylase a commercially available product or a product isolated from a microorganism may be used.
  • the microorganism as an isolation source may be a recombinant microorganism having isoamylase-producing ability or a mutant strain obtained by mutating a naturally-occurring microorganism in addition to a naturally-occurring microorganism.
  • microbial origin isoamylase is not particularly limited, for example, Mairoidesu Odoratasu (Myroides odoratus) and Pseudomonas Amiroderamosa (Pseudomonas amyloderamosa) can be used those derived from.
  • pullulanase a commercially available product or a product isolated from a microorganism may be used.
  • the microorganism as an isolation source may be a recombinant microorganism having pullulanase-producing ability or a mutant strain obtained by mutating a naturally-occurring microorganism in addition to a naturally-occurring microorganism.
  • microbial origin of pullulanase is not particularly limited, for example, Klebsiella pneumoniae (Klebseilla pneumoniae) and brevis (Bacillus brevis) can be used those derived from.
  • pullulanase “Amano” 3 manufactured by Amano Enzyme can be preferably used.
  • the branched megalosaccharide according to the present invention can be obtained in the reaction product. Whether a branched structure is introduced at the non-reducing end of the megalo sugar present in the reaction product is determined by ⁇ -amylase that hydrolyzes ⁇ -1,4-bonds in units of two sugars from the non-reducing end of maltodextrin or starch Can be confirmed by not undergoing hydrolysis.
  • the ⁇ -amylase that can be used is not particularly limited. For example, soybean-derived ⁇ -amylase can be used.
  • ⁇ -amylase As with ⁇ -amylase, it is possible to use maltodextrin or glucoamylase that hydrolyzes ⁇ -1,4-linkage from the non-reducing end of starch, but this enzyme is ⁇ -1,6- Since it also has a binding resolution, it is preferable to use ⁇ -amylase.
  • branched megalosaccharide in which a glucose residue is bonded to the non-reducing end by an ⁇ -1,3-glucoside bond can be produced.
  • the branched structure of the branched megalosaccharide includes a structure in which glucose is branched by ⁇ -1,3-bonds, a structure in which maltose is branched by ⁇ -1,3-bonds, and nigerose is formed by ⁇ -1,3-bonds.
  • a branched structure a structure in which maltotriose is branched by an ⁇ -1,3-bond, a structure in which maltosyl- ⁇ -1,3-glucose is branched by an ⁇ -1,3-bond, nigerosyl- ⁇ -1,4- Examples include a structure in which glucose is branched by an ⁇ -1,3-bond, and a structure in which nigerotriose is branched by an ⁇ -1,3-bond.
  • the branched structure is a glucan that is bonded to the non-reducing end of the linear glucan of the substrate by an ⁇ -1,3-linkage, and is a glucoside constituting the branched structure
  • the bond may be a glucan composed of ⁇ -1,4-bond and / or ⁇ -1,3-bond.
  • the branched megalosaccharide according to the present invention can be produced in a high yield, and in particular, a relatively high degree of polymerization with a degree of polymerization of 15 to 35. Therefore, it is possible to produce a highly branched megalosaccharide with high efficiency.
  • the branched structure of the branched megalosaccharide includes a structure in which glucose is branched by an ⁇ -1,6-bond, a structure in which maltose is branched by an ⁇ -1,6-bond, and an isomaltose in an ⁇ -1,6-bond.
  • Branched structure maltotriose branched by ⁇ -1,6-bond, isopanose branched by ⁇ -1,6-bond, panose branched by ⁇ -1,6-bond
  • An example is a structure in which maltotriose is branched by ⁇ -1,6-bonds.
  • the branched structure is a glucan that is bonded to the non-reducing end of the linear glucan of the substrate by an ⁇ -1,6-bond, and the glucoside constituting the branched structure
  • the bond may be a glucan having an ⁇ -1,4-bond and / or an ⁇ -1,6-bond.
  • ⁇ -glucosidase derived from Aspergillus niger is used, ⁇ -1,2-bonds and ⁇ -1,3-bonds may be included in the branched structure, although in very small amounts.
  • 6- ⁇ -glucosyltransferase When 6- ⁇ -glucosyltransferase is used as a glycosyltransferase, a branched megalosaccharide in which one molecule of glucose is bonded to the non-reducing end of a linear glucan by ⁇ -1,6-linkage is obtained.
  • dextrin dextranase When dextrin dextranase is used as a glycosyltransferase, a branched megalosaccharide in which a branched structure consisting of 1 to 6 glucose residues is bonded to the non-reducing end of a linear glucan by an ⁇ -1,6-bond is obtained. It is done.
  • This branched glucan is mainly composed of ⁇ -1,6-bonds, but may contain a few ⁇ -1,4-linkages.
  • a cyclic maltosyl maltose-producing enzyme is used as a glycosyltransferase
  • a branched megalosaccharide in which maltose or maltosyl- ⁇ -1,6-maltose is bonded to the non-reducing end of a linear glucan by an ⁇ -1,6-linkage Is obtained.
  • the product obtained by the enzymatic reaction can be directly used as a branched megalosaccharide product. Further, if necessary, the product obtained by the enzyme reaction is centrifuged or filtered to remove insoluble matters, and the water-soluble fraction is concentrated, whereby the target branched megalosaccharide solution according to the present invention is obtained. It can also be obtained. Or what was decolored with activated carbon as needed, and what removed the ionic component with the appropriate ion exchange resin may be concentrated. In the storage and subsequent use, it is preferable to concentrate the decolorized and ion-removed product until the water activity is such that the propagation of microorganisms does not become a problem. Or it can also be dried and obtained as a powder so that it may be easy to use depending on a use. For drying, a method such as freeze drying, spray drying or drum drying can be used. The dried product is desirably pulverized as necessary.
  • liquid sugars and powdered sugars containing 10 to 60% by mass, preferably 15 to 55% by mass, of the branched megalosaccharide according to the present invention per solid content.
  • powdered sugar shall be used with the meaning containing powdered rice cake.
  • the product obtained by the enzymatic reaction of the present invention usually contains a small amount of cyclodextrin, dextrin, oligosaccharide having a polymerization degree of 10 or less, or a mixture thereof, together with the branched megalosaccharide according to the present invention.
  • This product can be used as it is for foods and the like as described later, but if necessary, these components may be removed to obtain a purified branched megalosaccharide.
  • oligosaccharides having a polymerization degree of less than 5 may be removed from these products.
  • the product obtained by the enzyme reaction of the present invention has a branched structure derived from the starch raw material in the middle part of the linear glucan in addition to the non-reducing end, that is, the sugar residue other than the terminal residue.
  • the branched megalosaccharide is slightly contained, and such a branched megalosaccharide is also within the scope of the present invention.
  • the branched megalosaccharide according to the present invention thus obtained does not show turbidity due to aging compared to ordinary dextrin even when the solution is left at a low temperature or as a high-concentration solution having a solid content of about 75% by mass. It has the characteristic of having remarkable aging resistance. Moreover, it has the characteristics that the viscosity in the solution state of about 75 mass% of solid content is low and it is excellent in operativity. In addition, the content of low-molecular-weight oligosaccharide components such as glucose after saccharification reaction is small, and the varicella is also characterized by extremely low sweetness.
  • liquid sugars and powdered sugars containing branched oligosaccharides according to the present invention from which oligosaccharides having a polymerization degree of 4 or less have been removed have not only aging resistance but also low sweetness. And is advantageous in that it can be used for flavor improvement.
  • branched megalo sugar [ use for food]
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can be added to foods. Therefore, according to the present invention, a food additive comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same is provided.
  • the branched megalosaccharide according to the present invention and the food to which the liquid sugar and powdered sugar containing it can be added are not particularly limited.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same have a taste improving effect and a texture improving effect, and are characterized by low sweetness. Therefore, by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to the food, it is advantageous in that the flavor and texture of the food can be improved without impairing the original flavor of the food.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same also have an effect of uniformly freezing water in which the additive is dissolved or dispersed. Therefore, it is advantageous in that ice having a uniformly dispersed additive can be produced by adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same to water in which the additive is dissolved or dispersed. .
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it can also improve the shine and gloss of food. Therefore, by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it to the food, it is advantageous in that a product with improved shine and gloss can be provided.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can also suppress milk protein aggregation and precipitation. Therefore, by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to the milk protein-containing beverage, milk protein aggregation and precipitation are suppressed, and the milk protein-containing beverage is stabilized. Is advantageous in that it can be provided.
  • a person skilled in the art can appropriately select a branched megalosaccharide and liquid sugar and powdered sugar containing the same according to the present invention according to the method for producing the food. Even if it manufactures, it may mix
  • the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same can also be used by adding to pharmaceuticals. Therefore, according to the present invention, there are provided a pharmaceutical additive comprising the branched megalosaccharide according to the present invention, and liquid sugar and powdered sugar containing the same.
  • a pharmaceutical additive comprising the branched megalosaccharide according to the present invention, and liquid sugar and powdered sugar containing the same.
  • the medicinal product to which the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing it can be added and examples thereof include preparations for oral administration, preferably tablets, powders, granules, fine powders, and the like. Granules, liquids, syrups (including dry syrups), capsules, lozenges and the like.
  • the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same have a flavor improving effect. Therefore, by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it to a pharmaceutical product, the taste of the active ingredient and other additive ingredients can be corrected or masked, and can also be used as an excipient. Is advantageous.
  • composition methods and pharmaceutical additives as raw materials for pharmaceuticals are well known to those skilled in the art, and branched megalosaccharides according to the present invention and liquid sugars and powdered sugars containing them are used as active ingredients and other pharmaceutical additives. Can be produced in accordance with a conventional method.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can improve the flavor of foods and the like as shown in the formulation examples described later. Therefore, according to the present invention, there are provided a flavor-improving agent and a masking agent for preparations comprising the branched megalosaccharide according to the present invention, and liquid sugar and powdered sugar containing the same.
  • examples of flavor improvement include improvement of unpleasant taste and odor peculiar to foods and food additives, for example, reduction of bitterness and / or astringency (for example, reduction of bitterness and astringency of polyphenols).
  • Reduction of acidity e.g., mitigation of acidity derived from vinegar and acidulants
  • reduction of egg taste and / or taste e.g., reduction of unpleasant taste of preservatives such as gluconic acid
  • reduction of alcoholic feeling e.g., Reduction of alcohol feeling such as vodka and whiskey
  • reduction of acid odor and other unpleasant odor for example, reduction of glucono delta lactone odor (gluconic acid odor) and dextrin odor
  • Examples of the flavor improvement include enhancement of richness (for example, enhancement of fruit juice feeling and milky feeling).
  • Examples of the product whose flavor is improved by the flavor improving agent according to the present invention include the foods described above.
  • the taste corrected or masked in the pharmaceutical preparation includes an unpleasant taste (for example, bitter taste, astringency, sour taste, and savory taste) peculiar to active ingredients and other additives for pharmaceutical preparations.
  • an unpleasant taste for example, bitter taste, astringency, sour taste, and savory taste
  • the flavor improving agent according to the present invention can be preferably used as a bitterness and / or astringency reducing agent.
  • the flavor improving agent according to the present invention is preferably a bitterness and / or astringency of polyphenols. It can be used as a reducing agent.
  • Polyphenols mean components having a plurality of phenolic hydroxyl groups in the molecule, and examples thereof include catechin, anthocyanin, chlorogenic acid and the like. Polyphenols are known to have various physiological functions favorable to living organisms, but most polyphenols exhibit strong bitterness and astringency, making it difficult to ingest high concentrations. Since the branched megalosaccharide according to the present invention effectively reduces the bitterness and / or astringency caused by polyphenols, the flavor improving agent according to the present invention is advantageous in that it allows high-concentration intake of polyphenols.
  • the branched megalosaccharide according to the present invention is also low in sweetness, which is advantageous in that the flavor can be improved without impairing the original flavor of the food even when the flavor improver according to the present invention is added to the food.
  • a method for improving flavor and a method for producing food with improved flavor comprising adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same to food.
  • the bitter and / or astringent taste comprising the addition of the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to foods having a bitter and / or astringent taste.
  • a reduction method and a method for producing a food with reduced bitterness and / or astringency As described above, the branched megalosaccharide according to the present invention effectively reduces the bitterness and / or astringency derived from polyphenols. Therefore, as a food having a bitterness and / or astringency, a polyphenols-containing food is used. Can be mentioned.
  • the polyphenol-containing food is not particularly limited as long as it contains polyphenols, and foods to which polyphenols are added as well as foods originally containing polyphenols are also applicable.
  • the form of the food is not particularly limited, and includes not only solids but also semisolids and liquids.
  • Specific examples of polyphenol-containing foods include beverages such as tea beverages (green tea beverages, tea beverages, oolong tea beverages, etc.), fruit beverages, carbonated beverages, vegetable beverages, sports beverages, milk beverages, alcoholic beverages, and ice creams. , Jelly, mousse, candy, gum, filling, health food, supplements and the like.
  • the term “food product” is used to include beverages.
  • an unpleasant taste masking method or an unpleasant taste masked pharmaceutical comprising adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to the pharmaceutical.
  • a manufacturing method is provided.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can also improve the texture of foods and the like. Therefore, according to the present invention, there is provided a texture improving agent comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same.
  • examples of the texture improvement include suppression of a feeling of dryness, imparting a moist feeling, and maintaining a soft texture.
  • examples of the food whose texture is improved by the flavor improving agent according to the present invention include foods whose food texture is required to be improved, and examples of such foods include breads, cookies, and cooked rice. , Cakes, noodles, frozen foods.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it can be added to foods for the purpose of improving the texture, but it goes without saying that they may also be added for the purpose of improving the flavor.
  • the texture improving effect of preventing the puffiness is obtained, but the shelf life improver added to the cooked rice (for example, , Glucono delta lactone and gluconic acid) and the like can be reduced.
  • a method for improving texture and a method for producing food with improved texture comprising adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same to food. Provided.
  • the branched megalosaccharide and liquid sugar and powdered sugar containing the same according to the present invention can uniformly freeze water in which the additive is dissolved or dispersed. Therefore, according to the present invention, there is provided a uniform ice freezing agent comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same.
  • a uniform ice freezing agent comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same.
  • uniform freezing of ice means freezing water containing the additive in a state where the additive is uniformly dissolved or dispersed.
  • water in which an additive is dissolved or dispersed includes a solution in which components other than water, such as fruit juice itself and plant extract itself, are originally mixed.
  • the homogeneous ice is obtained, which comprises freezing the water in which the additive is uniformly dissolved or dispersed by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same.
  • a freezing method or a method for producing uniformly frozen ice is provided.
  • branched megalosaccharide according to the present invention By adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same, water can be frozen in a state where the additive is uniformly dissolved or dispersed. Therefore, for example, when the obtained ice is used as a frozen dessert, an iced dessert having a uniform color, texture, taste, etc. without unevenness and unevenness can be provided.
  • the ice uniform freezing agent according to the present invention is added to a beverage such as a fruit juice beverage or a sports beverage and frozen, a beverage that becomes a uniform solution even when partly thawed can be provided. .
  • the ice produced by the method for producing ice according to the present invention is used as it is or after processing, for example, ice confectionery such as sherbet, shaved ice, flake ice, ice candy, shake, sorbet; fruit juice drink, sports drink, amino acid drink Beverages such as jelly and yogurt.
  • ice confectionery such as sherbet, shaved ice, flake ice, ice candy, shake, sorbet
  • fruit juice drink, sports drink amino acid drink Beverages such as jelly and yogurt.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it can also improve the shine and gloss of food. Therefore, according to the present invention, there is provided a food shine and / or luster improving agent comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same.
  • meat is mentioned as a food in which the shimmering and / or improvement in gloss of the food is expected by the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same, and preferably chicken.
  • the method for improving the shine and / or shine of the food comprising adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to the food is improved.
  • a method for producing a prepared food for example, the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same may be applied to the food in advance, and the food may be baked. After the food is baked, the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same may be applied again and baked.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the teriyaki sauce may be added, and the food may be immersed in the sauce and then baked.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it may be directly sprayed on the surface of the food before baking.
  • the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same can be used by adding to a milk protein-containing beverage. Even when the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it are added to a milk protein-containing beverage, the milk protein does not aggregate and precipitate, and the milk protein can be stabilized in the beverage. That is, when the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it are used in a milk protein-containing beverage, aggregation and precipitation of milk protein can be avoided.
  • the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it have a lower osmotic pressure in the solution than monosaccharide, disaccharide, oligosaccharide and the like. Therefore, the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can be preferably used by being added to an enteral nutrition or oral nutrition containing milk protein.
  • milk protein in which aggregation and / or precipitation of milk protein is suppressed comprising adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same to a milk protein-containing beverage.
  • a method for producing a beverage containing beverage is provided.
  • the addition timing of the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same in these methods is not particularly limited, and may be added together with milk protein or separately from milk protein.
  • the food additive according to the present invention, the flavor improving agent, the texture improving agent, the ice uniform freezing agent, the shine and / or gloss improving agent, and the milk protein aggregation and / or precipitation inhibitor are added to the food.
  • a food product comprising an agglomeration and / or precipitation inhibitor is provided.
  • a food comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same.
  • the food provided may be a polyphenol-containing food, and specific examples and preferred examples are as described above.
  • branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same are used for polyphenols-containing foods, for example, when targeting foods and drinks that require sweetness, A composition having a strong sweetness such as sugar, isomerized sugar, and a high-intensity sweetener can be mixed and used.
  • a composition having a strong sweetness such as sugar, isomerized sugar, and a high-intensity sweetener can be mixed and used.
  • branched megalosaccharides alone or a mixture of branched megalosaccharides and polysaccharides, proteins, fats and the like.
  • a polyphenol-containing food containing 1.6 to 50% by mass of the branched megalosaccharide according to the present invention with respect to 0.13% by mass of the polyphenols.
  • the food to be provided may be a milk protein-containing beverage, more preferably an enteral nutrient or an oral nutrient containing milk protein.
  • the formulation additive and the formulation masking agent according to the present invention can be used in combination with a pharmaceutical product. Therefore, according to another aspect of the present invention, there is provided a pharmaceutical comprising the pharmaceutical additive according to the present invention and the pharmaceutical masking agent according to the present invention.
  • a medicinal product comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same.
  • the present invention will be specifically described based on the following examples, but the present invention is not limited to these examples.
  • the ratio per “solid content” or the content ratio of “solid content” means the ratio determined based on the mass of the solid component.
  • Test Example 1 Measurement of saccharifying enzyme activity 1-1: Activity measurement of cyclodextrin-forming enzyme Cyclodextrin-forming enzyme (CGTase) used in the saccharification reaction is shown below.
  • -CGTase derived from Paenibacillus sp . Alkaline CD amylase manufactured by Nagase ChemteX Corp.
  • -CGTase derived from Bacillus coagulans Neo CD amylase manufactured by Nagase ChemteX Corp.
  • CGTase derived from Bacillus mazelans Contizyme manufactured by Amano Enzyme
  • the enzyme reaction was performed by appropriately diluting water with 0.9 ml of 1% soluble starch (Nacalai Tesque) dissolved in 50 mM potassium phosphate buffer (pH 6.0). 0.1 ml of solution was added and held at 40 ° C. for 10 minutes. The reaction was stopped by adding 2.5 ml of 40 mM sodium hydroxide aqueous solution thereto. The produced ⁇ -cyclodextrin was measured by the phenolphthalein method.
  • ⁇ -glucosidase activity The ⁇ -glucosidase used in the saccharification reaction is shown below.
  • ⁇ -glucosidase derived from Aspergillus niger transglucosidase manufactured by Amano Enzyme, Amano Acremonium sp-derived ⁇ -glucosidase: manufactured by Kirin Foodtech Co., Ltd.
  • the enzyme reaction was dissolved in 50 mM sodium acetate buffer (pH 4.2). 20 ⁇ l of an enzyme solution appropriately diluted with 10 mM sodium acetate buffer (pH 4.2) containing 0.05% Triton X-100 was added to 80 ⁇ l of 25% maltose, and kept at 37 ° C.
  • the isoamylase used in the saccharification reaction is shown below.
  • -Isoamylase derived from Myroides odoratus GODO-FIA manufactured by Godo Shusei Co., Ltd.
  • Pseudomonas amyloderamosa-derived isoamylase Isoamylase produced by Hayashibara Bioscience Laboratories was prepared by adding 350 mg of 5 mg / ml waxy corn starch (Nippon Food Chemical Co., Ltd.) to 100 ⁇ l of 50 mM sodium acetate buffer solution (pH 6.0) containing 20 mM calcium chloride.
  • reaction stop solution 100 ⁇ l of an enzyme solution appropriately added with the same buffer was added to what was added and kept at 45 ° C. for 5 minutes, and kept at 45 ° C. for 15 minutes.
  • the reaction was stopped by adding 500 ⁇ l of a reaction lost iodine solution (a mixture of 2 ml of a solution consisting of 6.35 mg / ml iodine and 83 mg / ml potassium iodide and 8 ml of 0.1N hydrochloric acid).
  • the reaction stop solution was kept at room temperature for 15 minutes, and 10 ml of pure water was added thereto, and the absorbance at 610 nm was measured.
  • Test Example 2 Preparation of branched megalosaccharide The structure and physical properties of a reaction product obtained by allowing cyclodextrin-forming enzyme, ⁇ -glucosidase and debranching enzyme to act on starch liquefaction liquid were examined.
  • branched megalosaccharide was confirmed by the following method using the reaction solution. That is, 50 ⁇ L of 10 mg / mL ⁇ -amylase # 1500 (Nagase ChemteX) dissolved in 1 M sodium acetate buffer (pH 5.5) was added to 1 mL of sugar solution adjusted to 5% by mass, and the mixture was added at 55 ° C. for 1 hour. It was allowed to act to a certain extent and was inactivated by boiling. This was desalted with Amberlite MB4 (organo) and then filtered through a 0.45 ⁇ m filter for high performance liquid chromatography (HPLC). As HPLC conditions, MCI GEL CK02AS was used in the column.
  • the mobile phase was ultrapure water
  • the column temperature was 85 ° C.
  • the flow rate was 1.0 ml / min.
  • about 20 ⁇ L was subjected to chromatography. Under any condition, the content of each polymerization degree component can be determined from the peak area of the obtained chromatogram, and the content of carbohydrates having a polymerization degree of 11 or more was calculated as the branched megalosaccharide content.
  • a branched megalosaccharide having a branched structure at the non-reducing end can be obtained by allowing a cyclodextrin-forming enzyme to act on a starch liquefaction solution in the presence of ⁇ -glucosidase.
  • components other than the components in the table are low-molecular oligosaccharides having a degree of polymerization of 10 or less, and dextrin refers to a polymer component having a degree of polymerization of more than 35.
  • Test Example 3 Confirmation of Aging Resistance of Branched Megalosaccharide
  • a syrup containing a branched megalosaccharide was prepared and checked for turbidity due to aging. was used to evaluate aging resistance.
  • the resulting saccharification reaction solution (equivalent to syrup in Production Example 5 described later) was heated at 80 ° C. for 30 minutes, and ⁇ -amylase preparation Christase L1 manufactured by Daiwa Kasei Co., Ltd. was 0.005% per solid content. The reaction was continued until the iodine reaction disappeared. The enzyme was inactivated by adjusting the pH to 4.0, and then neutral sugar was purified by filtration, ion exchange purification, and activated carbon treatment. Amberlite MB3 was used for ion exchange purification, and purified white birch (Kirin Foodtech) was used for activated carbon treatment. The liquid sugar obtained by concentrating the purified sugar liquid to Bx75 was kept at room temperature for 1 month. Table 3 shows the sugar composition of the resulting sugar solution.
  • An ⁇ -amylase degradation product (corn syrup) of starch was used as a comparative control of storage stability with branched megalosaccharide-containing syrup. That is, 22.5% by mass of DE 6.5 corn starch liquefied liquid was obtained by adding 0.005% of the Christase L1 to the solid content, 0.04% of the pullulanase “Amano” 3 to the solid content, and Myloides odolatas isoamylase to the solid. 400 units were added per gram per minute and saccharified for 36 hours at pH 6.0 and 53 ° C. After heating this to 80 ° C., Christase L1 was additionally added to the solid content by 0.005% and held until the iodine reaction disappeared.
  • Neutral sugars were purified and concentrated in the same manner as above from the resulting reaction solution.
  • Table 3 shows the sugar composition of the resulting sugar solution.
  • components other than the components in the table are low-molecular oligosaccharides having a polymerization degree of 10 or less.
  • Megalosaccharide refers to a saccharide having a polymerization degree of 11 to 35, and the content in parentheses indicates branched megalosaccharide content. Refers to a polymer component having a degree exceeding 35.
  • Test Example 4 Measurement of Viscosity of Branched Megalosaccharide-Containing Syrup
  • the viscosity of the branched megalosaccharide-containing syrup produced in Test Example 3 was measured and compared with the existing oligosaccharide syrup. That is, the viscosity at 10 ° C. to 60 ° C. of the branched megalosaccharide-containing syrup prepared in Test Example 3 was measured.
  • the concentration of syrup was Bx70.
  • the viscosity was measured using VISCOMETER TVB-10 manufactured by Toki Sangyo Co., Ltd., the rotation speed of the rotor was 60 rpm, and the measurement time was 40 seconds.
  • a 300 mL sample was placed in a 300 mL tall beaker and used as a measurement sample.
  • the viscosity of the branched megalosaccharide-containing syrup at each temperature was higher than that of Fuji Oligo G67 manufactured by Nippon Shokuhin Kako Co., Ltd., which is an existing polymer maltooligosaccharide-containing syrup, the viscosity is such that there is no problem in operability even at a solid content of 70%. Met.
  • Test Example 5 Measurement of Sweetness of Branched Megalosaccharide-Containing Syrup
  • the sweetness of the branched megalosaccharide-containing syrup produced in Test Example 3 was measured by Pauli's whole series method (starch sugar related industrial analysis method) Measured according to Starch Sugar Technical Committee, pages 61-62). That is, the sweetness of the sugar aqueous solution prepared at intervals of 0.2% in the range of 0.6 to 1.4% and the branched megalosaccharide-containing syrup having a solid content of 10% is “strong”, “equal”, “weak”. Ten panelists evaluated. Based on the obtained test results, the sweetness degree was calculated by the following formula.
  • S Em / N ⁇ 100
  • Do lowest concentration of standard sugar solution
  • Du highest concentration of standard sugar solution
  • i concentration interval of standard sugar solution
  • ⁇ gr number of people judged as “strong”
  • ⁇ kl number of people judged as “weak”
  • n Number of panelists
  • S sweetness
  • N sample concentration (%)
  • the sweetness of the branched megalosaccharide-containing syrup was 10.7, and it was revealed that the sweetness was extremely low compared with the existing liquid sugar.
  • Production Example 1 Production of branched megalosaccharide (1)
  • Production of branched megalosaccharide 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Saccharification for 72 hours by adding 100 units per gram of odoratus isoamylase, 0.01% of pullulanase “Amano” 3 per solid, and 3.75 units of ⁇ -glucosidase of Aspergillus niger per gram of solid did. This was heated to 80 ° C., and Christase L1 was added in an amount of 0.005% to the solid content until the iodine reaction disappeared.
  • Activated charcoal was added to the inactivated enzyme at pH 4.0 and decolorized according to a conventional method, followed by filtration through diatomaceous earth to obtain a filtrate. Although this was made into a usual method, the ionic component was removed with the ion exchange resin, and the neutral sugar was obtained. This was decolorized by performing the activated carbon treatment again in the same manner as above, and filtered through diatomaceous earth to obtain a filtrate. This was concentrated under reduced pressure to 75% solids. By the above operation, a branched megalosaccharide-containing syrup was obtained at a yield of about 90% based on the solid content.
  • This product contains 20.3% of megalosaccharide per solid, and when the content of branched megalosaccharide was measured by ⁇ -amylase treatment, branched megalosaccharide having a degree of polymerization of 11 to 35 per solid was measured. It contained 17.9%.
  • the aqueous layer was removed, and 1 mL of water was newly added and stirred for 5 times, and the chloroform layer was recovered.
  • the obtained sample was subjected to gas chromatography to analyze the partially methylated monosaccharide alditol acetate.
  • TC-17 (0.25 mm ⁇ 30 m; manufactured by GL Science) was used for the column, the column temperature was kept at 50 ° C. for 1 minute, and then the column temperature was increased to 280 ° C. by 10 ° C. per minute.
  • the sample injection temperature was 300 ° C.
  • Detection was performed by FID, and the detection temperature was 300 ° C.
  • Helium was used as the carrier gas, and the flow rate was 2.5 mL / min.
  • 1,5-di-O-acetyl-2,3,4,6-tetra-O was obtained as a partially methylated derivative derived from branched megalosaccharide obtained using ⁇ -glucosidase derived from Aspergillus niger.
  • -Methylhexitol, 1,5,6-tri-O-acetyl-2,3,4-tri-O-methylhexitol and 1,4,5-tri-O-acetyl-2,3,6 , -Tri-O-methylhexitol was detected. This revealed that the branched structure of the branched megalosaccharide obtained in Production Example 1 (1) is an ⁇ -1,6-bond.
  • the reaction solution obtained by filtration through a 0.45 ⁇ m filter was concentrated and used as a fraction source. This was subjected to carbon-celite column chromatography. In this chromatography, sugar was eluted with a linear concentration gradient of 0-3% n-butanol.
  • the obtained pentasaccharide fraction was desalted with an ion exchange resin, and then filtered through a 0.45 ⁇ m filter and freeze-dried. Of this, 5 mg was subjected to methylation analysis as described above.
  • Production Example 2 Production of branched megalosaccharide (2) A 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus coagulans cyclodextrin-producing enzyme was paired with 1 unit per gram of solid content and Myloides odoratus isoamylase. Saccharification was performed for 60 hours by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 3.75 units of ⁇ -glucosidase of Aspergillus niger per gram of solid content.
  • Production Example 3 Production of branched megalosaccharide (3) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus stearothermophilus cyclodextrin-forming enzyme was added to 1 unit per gram of solid content, and Myloides odoratus isoform. Saccharification was performed by adding 100 units per gram of amylase, 0.01% of pullulanase “Amano” 3 per solid, and 3.75 units of ⁇ -glucosidase of Aspergillus niger per gram of solid for 60 hours.
  • Production Example 4 Production of branched megalosaccharide (4) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus mazellans cyclodextrin-forming enzyme was paired with 1 unit per gram of solids and Myloides odoratus isoamylase. Saccharification was performed for 60 hours by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 3.75 units of ⁇ -glucosidase of Aspergillus niger per gram of solid content.
  • Production Example 5 Production of branched megalosaccharide (5)
  • Manufacture of branched megalosaccharide 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. 100 mg / g of Myloides odoratus isoamylase, 0.01% of pullulanase “Amano” 3 per g of solid, and 0.65 unit of ⁇ -glucosidase of Acremonium sp. Saccharified for 60 hours.
  • the subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content.
  • the product contained 81.3% of megalosaccharide per solid and 56.1% of branched megalosaccharide per solid.
  • 1,5-di-O-acetyl-2,3,4,6-tetra- was obtained as a partially methylated derivative derived from branched megalosaccharide obtained by using ⁇ -glucosidase derived from Acremonium sp.
  • O-methylhexitol, 1,3,5-tri-O-acetyl-2,4,6-tri-O-methylhexitol and 1,4,5-tri-O-acetyl-2,3 6-Tri-O-methylhexitol was detected.
  • This revealed that the branched structure of the branched megalosaccharide obtained in Production Example 5 (1) is an ⁇ -1,3-linkage.
  • the reaction solution obtained by filtration through a 0.45 ⁇ m filter was concentrated and used as a fraction source. This was subjected to carbon-celite column chromatography. In this chromatography, sugar was eluted with a linear concentration gradient of 0-3% n-butanol.
  • the obtained pentasaccharide fraction was desalted with an ion exchange resin, and then filtered through a 0.45 ⁇ m filter and freeze-dried. Of this, 5 mg was subjected to methylation analysis as described above.
  • Production Example 6 Production of branched megalosaccharide (6) A 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus coagulans cyclodextrin-producing enzyme was paired with 1 unit per gram of solid content and Myloides odoratus isoamylase. Saccharification was carried out by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 0.65 units of Acremonium sp. ⁇ -glucosidase per gram of solid content for 60 hours.
  • Production Example 7 Production of branched megalosaccharide (7) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus stearothermophilus cyclodextrin-forming enzyme was added to 1 unit per gram of solid content, and Myloides odoratus isoform. Saccharification was performed by adding 100 units per gram of amylase, 0.01% of pullulanase “Amano” 3 per solid, and 0.65 units of Acremonium sp. ⁇ -glucosidase per gram of solid for 60 hours.
  • Production Example 8 Production of branched megalosaccharide (8) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus mazellans cyclodextrin-forming enzyme was paired with 1 unit per gram of solids and Myloides odoratus isoamylase. Saccharification was carried out by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 0.65 units of Acremonium sp. ⁇ -glucosidase per gram of solid content for 60 hours.
  • Production Example 9 Production of branched megalosaccharide (9) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and pH 6.0, to which Paenibacillus sp. Cyclodextrin-producing enzyme was paired with 2 units per gram of solid content and Pseudomonas amylodelamosa isoamylase. Saccharification was carried out for 60 hours by adding 200 units per gram of solid content, 0.02% of pullulanase “Amano” 3 per solid content, and 3.75 units of ⁇ -glucosidase of Aspergillus niger per gram of solid content.
  • Production Example 10 Production of branched megalosaccharide (10) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-producing enzyme was paired with 1 unit per gram of solid content and Pseudomonas amylodelamosa isoamylase was paired. Saccharification was carried out by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 0.65 units of Acremonium sp. ⁇ -glucosidase per gram of solid content for 60 hours.
  • Production Example 11 Production of Branched Megalosaccharide (11) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-forming enzyme was paired with 1 unit per 1 g of solid content and pullulanase “Amano” 3 was paired. 0.01% per solid, Aspergillus niger ⁇ -glucosidase was added to 3.75 units per gram of solid and saccharified for 60 hours.
  • Production Example 12 Production of branched megalosaccharide (12) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-producing enzyme was paired with 1 unit per 1 g of solid content, and pullulanase “Amano” 3 was paired. Saccharification was carried out for 60 hours by adding 0.65 unit of ⁇ -glucosidase of 0.01% per solid and 0.01% Acremonium sp.
  • Production Example 13 Production of branched megalosaccharide (13) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-forming enzyme was added to 1 unit per gram of solid and Aspergillus niger ⁇ -glucosidase. Saccharification was carried out for 60 hours by adding 3.75 units per gram of the solid content. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. The product contained 17.3% of megalosaccharide per solid and 17.0% of branched megalosaccharide per solid.
  • Production Example 14 Production of Branched Megalosaccharide (14) 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-producing enzyme was 1 unit per gram of solid content, Acremonium sp ⁇ -glucosidase Was saccharified for 60 hours by adding 0.65 units per gram of the solid content. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. In addition, this product contained 32.6% of megalo sugar per solid content, and contained 29.2% of branched megalo sugar per solid content.
  • Production Example 15 Production of powdered koji containing branched megalo sugar (1)
  • the sugar solution obtained in Production Example 1 was pulverized using an L-12 type spray dryer (atomizer type: Okawara Chemical).
  • the spray dryer was operated at an inlet set temperature of 160 ° C., an outlet set temperature of 120 ° C., an atomizer rotational speed of 10,000 rpm, a cyclone differential pressure of about 70 mmHg, and a liquid supply amount of 5 L / h.
  • a powdered branched megalosaccharide-containing powder cake with a recovery rate of about 95% was obtained.
  • Production Example 16 Production of branched megalosaccharide-containing flour (2) The sugar solution obtained in Production Example 5 was spray-dried in the same manner as in Production Example 15 using an L-12 type spray dryer. By this operation, a powdered branched megalosaccharide-containing powder cake with a recovery rate of about 95% was obtained.
  • Production Example 17 Production of Branched Megalosaccharide Fraction Product
  • a gel filtration column (TOYOPEARL HW-40S, ⁇ 5.0 cm ⁇ 87 cm, prepared by preparing the branched megalosaccharide-containing syrup of Production Example 1 to about Brix 50 and heating to 60 ° C. Elution was carried out using purified water at a flow rate of 34 to 48 ml / h.
  • the saccharide composition of the eluate was analyzed by high performance liquid chromatography, and the fraction of branched sugars below tetrasaccharide (high molecular fraction) and the fraction of 0.5% or higher (low molecular content fraction). Min) was collected.
  • the collected fraction was concentrated to about Brix 30 by drying under reduced pressure. By repeating the above operation 11 times, 23 g of a polymer fraction and 39 g of a low-molecule-containing fraction were obtained in terms of solid content.
  • a branched megalosaccharide reduced product was prepared using NaBH 4 . That is, after cooling 10 minutes branched cytomegalovirus sugars adjusted to Bx10 (Preparation Example 5) solution 5ml in ice, and dissolved by the NaBH 4 by about 20mg added 10 times. Two drops of aqueous ammonia were added to this solution to adjust the pH to 10 or more, and then a reduction reaction was performed at room temperature for 3 hours. After the reaction, the remaining NaBH 4 was decomposed by adding Amberlite MB-4 (organo) until hydrogen generation and the color change of the resin disappeared. The liquid layer was collected while washing the resin with water and dried using a rotary evaporator. 1 ml of methanol was added to this and dissolved, and again dried. This operation was repeated 5 times. Finally, the product dissolved in purified water was used as a reduced product solution.
  • the DE of the obtained reduced product was measured according to a modified somogy method (starch sugar related industrial analysis method (Food Chemical Newspaper Co., Ltd.) (issued on November 1, 1991), pages 11 to 13). Met. The DE before the reduction was 10.2, and it was confirmed that the reducing end was completely reduced by the reduction reaction. In addition, the composition of the degree of polymerization did not change before and after the reduction reaction, and no glucoside bond breakage was observed (data not shown).
  • Formulation Example 1 Green tea beverage A green tea beverage was produced according to the formulation shown in Table 4 (mass ratio based on 100 as a whole).
  • the syrup of Production Example 5 was used as the branched megalosaccharide syrup.
  • Celldex SL-20, Celldex B-100, and Fujioligo G67 were manufactured by Nippon Shokuhin Kako Co., Ltd. (hereinafter the same).
  • Celdex SL-20 is a liquid or powder product containing 20% cyclodextrin
  • Celdex B-100 is crystalline ⁇ -cyclodextrin.
  • Fuji oligo G67 is syrup mainly composed of maltohexaose and maltoheptaose.
  • the addition ratio of the syrup-like raw material was calculated based on the mass of the solid content (hereinafter the same).
  • Table 5 shows the sugar composition and sweetness of the blended megalosaccharide syrup and Fujioligo G67.
  • the sugar composition was analyzed using HPLC. The analysis conditions were the same as in Test Example 2. The sweetness level was measured by the method described in Test Example 5.
  • the bitterness and astringency reduction effects in the table are shown as evaluation results of very effective ( ⁇ ), effective ( ⁇ ), almost no effect ( ⁇ ), no effect ( ⁇ ), and sweetness is strong. ( ⁇ ), slightly sweet ( ⁇ ), not sweet ( ⁇ ), umami is umami ( ⁇ ), umami is a little ( ⁇ ), umami is low ( ⁇ ), umami is weak (x), overall evaluation The evaluation results were very good ( ⁇ ⁇ ), good ( ⁇ ⁇ ), not very good ( ⁇ ), and bad ( ⁇ ).
  • Formulation Example 2 High-concentration polyphenols-containing green tea beverage A high-concentration polyphenol-containing green tea beverage was produced according to the formulation shown in Table 7 (mass ratio when the whole was 100). As the branched megalosaccharide syrup, the syrup of Production Example 5 was used. Polyphenon CH manufactured by Mitsui Norin Co., Ltd. was used (hereinafter the same).
  • Ten panelists performed sensory evaluation of the produced tea beverage and evaluated the balance of bitterness / astringency and taste. Table 8 shows the evaluation results.
  • the bitterness and astringency reduction effects in the table are shown as evaluation results of very effective ( ⁇ ), effective ( ⁇ ), almost no effect ( ⁇ ), no effect ( ⁇ ), and sweetness is strong. ( ⁇ ), slightly sweet ( ⁇ ), not sweet ( ⁇ ), umami is umami ( ⁇ ), umami is a little ( ⁇ ), umami is low ( ⁇ ), umami is weak (x), overall evaluation The evaluation results were very good ( ⁇ ⁇ ), good ( ⁇ ⁇ ), not very good ( ⁇ ), and bad ( ⁇ ).
  • the bitterness and astringency of polyphenols can be reduced, and umami can be imparted. Furthermore, these effects were obtained without breaking the balance of tea taste.
  • Formulation Example 3 Black Tea Beverage A black tea beverage was produced according to the formulation shown in Table 9 (mass ratio based on 100 as a whole).
  • the black tea used was a clear black tea extract obtained by adding 200 ml of hot water (95 ° C.) to 2 g of black tea leaves and performing extraction for 2 minutes, followed by suction filtration after extraction.
  • the syrup of Production Example 5 was used as the branched megalosaccharide syrup.
  • bitterness and astringency reduction effects in the table are shown as evaluation results of very effective ( ⁇ ), effective ( ⁇ ), almost no effect ( ⁇ ), no effect ( ⁇ ).
  • the evaluation results were very good ( ⁇ ), good ( ⁇ ), not very good ( ⁇ ), and bad ( ⁇ ).
  • Formulation Example 4 Green tea jelly Green tea jelly was prepared in a conventional manner with the formulation shown in Table 11 (mass ratio based on 100 as a whole). As the branched megalosaccharide syrup, the syrup of Production Example 5 was used.
  • bitterness and astringency reduction effects in the table are shown as evaluation results of very effective ( ⁇ ), effective ( ⁇ ), almost no effect ( ⁇ ), no effect ( ⁇ ). About ((circle)), there is no richness (x), and the balance of the taste was shown by the evaluation result of very good ((double-circle)), good ((circle)), not so good ((triangle
  • Formulation Example 5 Green tea beverage A green tea beverage was produced according to the formulation shown in Table 13 (mass ratio based on 100 as a whole). As the branched megalosaccharide syrup, the branched megalosaccharide syrup fraction produced in Production Example 17 was used.
  • the analysis results of the sugar composition of the blended megalosaccharide syrup fraction and Fujioligo G67 were as follows.
  • the sugar composition was analyzed using HPLC.
  • the analysis conditions were the same as in Test Example 2.
  • the sensory evaluation of the prepared green tea beverage was performed by 23 panelists, and the balance of bitterness / astringency and taste was evaluated. The evaluation results are shown in Table 15.
  • the bitterness and astringency reduction effects in the table are shown as evaluation results of very effective ( ⁇ ), effective ( ⁇ ), almost no effect ( ⁇ ), no effect ( ⁇ ), and sweetness is strong. ( ⁇ ), slightly sweet ( ⁇ ), not sweet ( ⁇ ), umami is umami ( ⁇ ), umami is slightly ( ⁇ ), umami is low ( ⁇ ), umami is not ( ⁇ ), balance of taste The evaluation results were very good ( ⁇ ), good ( ⁇ ), not very good ( ⁇ ), and bad ( ⁇ ).
  • Japanese Patent Application Laid-Open No. 2006-280254 discloses a method for suppressing the bitterness and astringency of catechin using a 3-4 saccharide having a branched structure as an active ingredient. Even when the sugar syrup fraction was used, the bitterness / astringency reduction effect was recognized. Further, the use of the branched megalosaccharide syrup according to the present invention, which contains almost no 3-4 saccharides, gave a high umami imparting effect.
  • Formulation Example 6 High concentration polyphenols-containing green tea beverage A high concentration polyphenols-containing green tea beverage was produced according to the formulation shown in Table 16 (mass ratio when the whole was 100). As the branched megalosaccharide syrup, the branched megalosaccharide syrup fraction produced in Production Example 17 was used.
  • bitterness and astringency reduction effects in the table are shown as evaluation results of very effective ( ⁇ ), effective ( ⁇ ), almost no effect ( ⁇ ), no effect ( ⁇ ).
  • the evaluation results were very good ( ⁇ ), good ( ⁇ ), not very good ( ⁇ ), and bad ( ⁇ ).
  • Formulation Example 7 Black tea beverage A black tea beverage was produced according to the formulation shown in Table 18 (mass ratio based on 100 as a whole).
  • the black tea used was a clear black tea extract obtained by adding 200 ml of hot water (95 ° C.) to 2 g of black tea leaves and performing extraction for 2 minutes, followed by suction filtration after extraction.
  • As the branched megalosaccharide syrup the branched megalosaccharide syrup fraction produced in Production Example 17 was used.
  • bitterness and astringency reduction effects in the table are shown as evaluation results of very effective ( ⁇ ), effective ( ⁇ ), almost no effect ( ⁇ ), no effect ( ⁇ ).
  • the evaluation results were very good ( ⁇ ), good ( ⁇ ), not very good ( ⁇ ), and bad ( ⁇ ).
  • Formulation Example 8 Green tea jelly Green tea jelly was prepared in a conventional manner with the formulation shown in Table 20 (mass ratio based on 100 as a whole). As the branched megalosaccharide syrup, the branched megalosaccharide syrup fraction produced in Production Example 17 was used.
  • the sensory evaluation of the produced green tea jelly was performed by 10 panelists, and the balance of bitterness / astringency and taste was evaluated. The evaluation results are shown in Table 21.
  • bitterness and astringency reduction effects in the table are shown as evaluation results of very effective ( ⁇ ), effective ( ⁇ ), almost no effect ( ⁇ ), no effect ( ⁇ ), and richness is effective
  • evaluation results of ( ⁇ ), no effect ( ⁇ ), and balance of taste were shown as evaluation results of very good ( ⁇ ), good ( ⁇ ), not very good ( ⁇ ), and bad ( ⁇ ).
  • the bitterness and astringency of polyphenols were reduced by using Fujioligo G67, Celdex SL-20, or Celdex B-100.
  • Fujioligo G67 was weaker than that of branched sugars.
  • Celdex SL-20 and Celdex B-100 reduced bitterness and astringency as much as or more than the branched megalosaccharide syrup, but the green tea flavor became difficult to feel.
  • the branched megalosaccharide syrup fraction according to the present invention when used, it was possible to suppress the bitterness and astringency of polyphenols. In addition, richness can be imparted, and the use of the branched megalosaccharide syrup fraction according to the present invention did not impair the overall taste balance.
  • Formulation Example 9 Fruit juice beverage A fruit juice beverage was prepared by blending 1% (w / w) (in terms of solid content) of the branched megalosaccharide syrup prepared in Production Example 1 with respect to a commercially available low fruit juice beverage. As a result, compared to the non-added group, the richness increased in the branched megalosaccharide added group, and it was possible to give depth to the fruit juice drink. Branched megalosaccharide can be advantageously used as a rich taste enhancer even in a low fruit juice beverage.
  • Formulation Example 10 High-sweetness sweetener-containing beverage Acesulfame K 0.05% (w / w) or aspartame 0.03% (w / w) -containing beverage 1% of the branched megalosaccharide syrup prepared in Production Example 1 A blended beverage (w / w) (solid content conversion) was prepared. The subtle sweetness of the aftertaste of branched megalo sugar worked effectively, and the bitterness of the aftertaste of high-intensity sweeteners was improved, and it was possible to make a drink that was comfortable to drink. Thus, the branched megalosaccharide according to the present invention can be advantageously used as a taste improving agent for high-intensity sweeteners.
  • Formulation Example 11 With respect to 100 parts by mass of the frozen fried egg whole eggs, 7 parts by mass of the branched megalosugar syrup prepared in Production Example 1 (in terms of solid content), 1.5 parts by mass of the modified starch, and 30 parts by mass of the soup stock are mixed.
  • Fried eggs were prepared by a conventional method. It was frozen with a quick freezer and thawed at room temperature after 2 weeks. Since this product contains a branched megalosaccharide having excellent aging resistance, it was kept fresh even after freezing and thawing. That is, it was confirmed that the branched megalosaccharide according to the present invention has an effect of maintaining freshness after freezing and thawing.
  • Formulation Example 12 2.2 parts by mass of each anti-powder yeast, 0.1 parts by mass of yeast food, 2 parts by mass of sugar, 3 parts by mass of the branched megalosaccharide syrup prepared in Production Example 1 (100 parts by mass of bread crumbs ) Solid content conversion), 2 parts by mass of sodium chloride, 2 parts by mass of skim milk powder, 6 parts by mass of fats and oils, and 67 parts by mass of water were used to prepare bread using a medium seed method. Since this product contains a branched megalosaccharide with excellent aging resistance, a bread with a soft texture was obtained. That is, it was confirmed that the branched megalosaccharide according to the present invention has an effect of giving a soft texture to food.
  • Formulation Example 13 17 parts by mass of soft cookie oil / fat, 50 parts by mass of branched megalosugar syrup (solid equivalent) prepared in Production Example 1 vs. powder, 60 parts by mass of sugar vs. powder, 50 parts by mass of whole egg Were sequentially added and mixed. After adding 0.2 parts by weight of vanilla oil to powder, 40 parts by weight of skim milk powder and powder, 100 sieved wheat flour and 0.9 parts by weight of baking powder were mixed. Firing was carried out for 13 minutes at 195 ° C. on the top and 170 ° C. on the bottom. Since this product contains a branched megalosaccharide with excellent aging resistance, it can retain a moist texture for a long period of time. That is, it was confirmed that the branched megalosaccharide according to the present invention has an effect of imparting a moist texture to food.
  • Formulation Example 14 Ice uniformity improvement / promotion effect 4% (w / w) of branched megalosaccharide syrup prepared in Production Example 5 (in terms of solid content), 5% of sugar (w / w), and 0 of pigment .4% (w / w) (Sun Red No. 5F (manufactured by San-Ei Gen FFI Co., Ltd.)) was added to prepare a mixed aqueous solution. This was frozen overnight in a ⁇ 30 ° C. freezer. Sampling was carried out over time at the time of melting at room temperature, and the elution state of branched megalosaccharide and sugar was confirmed by high performance liquid chromatography (HPLC).
  • HPLC high performance liquid chromatography
  • Formulation Example 15 Vinegar Beverage Apple Vinegar 8% (w / w), Fructose Glucose Liquid Sugar (manufactured by Nippon Shokuhin Kako Co., Ltd.) 8% (w / w) (in terms of solid content) and Branched Megalosaccharide Syrup
  • blended 1% (w / w) (solid content conversion) was evaluated. Since this product contains a branched megalosaccharide that has an excellent acidity-reducing effect, the acidity is softened and the drinking comfort can be improved. That is, it was confirmed that the branched megalosaccharide according to the present invention has a sour taste reducing effect.
  • Formulation Example 16 Mixing 154.5 parts by mass of water, 0.75 parts by mass of gluconodeltalactone and 2 parts by mass of the branched megalosaccharide syrup prepared in Production Example 1 (in terms of solid content) with 100 parts by mass of aseptic cooked rice, I cooked rice. Since it contains a branched megalosaccharide that has an excellent acid odor suppressing effect, the acid odor derived from gluconodeltalactone after cooking or during reheating was reduced. That is, the branched megalosaccharide according to the present invention has an effect of reducing the acid odor, and can be used as an acid odor reducing agent for a preservative.
  • Formulation Example 17 Fructose-glucose liquid sugar 4% (w / v) (in terms of solid content) and Production Example 5 so that the alcohol concentration is 6% (v / v) using Nikolai vodka (Kirin Seagram Co., Ltd.)
  • the alcoholic beverage blended with 1% (w / v) branched megalosaccharide syrup (converted to solid content) prepared in the above was evaluated. Since this product contains branched megalosaccharide, which is excellent in reducing alcohol feeling, the taste of alcohol is reduced and the drinking comfort is improved. That is, the branched megalosaccharide according to the present invention can be advantageously used as a taste improving agent for alcoholic beverages.
  • Formulation Example 18 Milk Beverage Milk 30% (w / w), Fructose Glucose Liquid Sugar 4% (w / w) (Solid Content) and Branched Megalosaccharide Syrup 1% (w / w) Prepared in Production Example 5 ( A solid beverage) was mixed to prepare a milk beverage. Compared to the additive-free zone, this product has improved milky taste and has a rich beverage. That is, the branched megalosaccharide by this invention can be used for the food-drinks containing milk for the purpose of the improvement of a milky taste.
  • Formulation Example 19 Jelly dextrin (TK-16 (manufactured by Matsutani Chemical Industry)) 20% (w / w), fructose-glucose liquid sugar 8% (in terms of solid content) (manufactured by Nippon Shokuhin Kako), cool agar (manufactured by Nitta Gelatin) 2% (w / w), tri-Na tricitrate 0.1% (w / w), citric acid 0.17% (w / w) and 5% of branched megalosaccharide syrup prepared in Production Example 5 (w / w) ) (In terms of solid content) was used to prepare a jelly.
  • TK-16 manufactured by Matsutani Chemical Industry
  • TK-16 is a powdery dextrin having a DE of 16-19. Since this product contains a branched megalosaccharide that is excellent in reducing the powdery odor of dextrin, the dextrin odor was alleviated even in jelly using a large amount of dextrin as in this test section. That is, the branched megalosaccharide according to the present invention can be advantageously used even when the food or drink contains a large amount of dextrin.
  • Blending Example 20 Teriyaki Sauce A Teriyaki sauce was prepared according to the blending shown in Table 22 (mass ratio when water was 200). As the branched megalosaccharide syrup, the syrup of Production Example 1 was used. Fuji oligo G67 is syrup mainly composed of maltohexaose and maltoheptaose. Chicken thighs were used and soaked overnight in a sauce that was 30% (w / w) of the weight of the meat. The chicken was steam-heated until it reached a central temperature of 70 ° C., and then poured again into the sauce, and then baked on a hot plate at 180 ° C. for 1.5 minutes on each side.
  • the teriyaki chicken produced by 10 panelists was subjected to sensory evaluation, and shine and gloss were evaluated. The evaluation results are shown in Table 23.
  • Formulation Example 21 Beverage containing milk protein (1)
  • a milk protein-containing beverage was produced according to the following procedure with the formulation shown in Table 24 (mass ratio when the whole was 40). That is, sugar or dextrin and skim milk powder shown in Table 24 were dissolved and mixed and stirred. Next, the pH was adjusted to 6.4 using citric acid, tri-Na citrate, and ascorbic acid. This aqueous solution was filled into a 50 ml Falcon tube, sealed and subjected to retort sterilization at 121 ° C. and 1.2 kgf / cm 2 (using HIGH-PRESSURE STEAM STERILIZER / BS-325 manufactured by Tommy Seiko Co., Ltd.). .
  • the syrup obtained in Production Example 5 was used as the branched megalosaccharide syrup.
  • Dextrin is Paindex # 2 (DE approx. 11 manufactured by Matsutani Chemical Industry Co., Ltd.), Paindex # 100 (DE approx. 2-5 manufactured by Matsutani Chemical Industry Co., Ltd.), Fujistar 5V (DE approx. 5 manufactured by Nippon Shokuhin Kako), Fujister 12V (DE About 12 (made by Nippon Food Chemicals), Paindex # 4 (DE about 19; made by Matsutani Chemical Industry), TK-16 (DE about 16-19 made by Matsutani Chemical Industry), cluster dextrin (DE about 2 made by Nippon Food Chemicals) did.
  • the aggregation state of milk protein in the beverage produced as described above was visually confirmed, and a stability test was performed.
  • the results are shown in Table 25.
  • the protein stability in the table is indicated by the evaluation results of stability ( ⁇ ), slightly stable ( ⁇ ), and unstable ( ⁇ ).
  • Formulation Example 22 Beverage containing milk protein (2) A milk protein-containing beverage was produced according to the following procedure with the formulation shown in Table 26 (mass ratio when the whole was 200). That is, sugar or dextrin and skim milk powder shown in Table 26 were dissolved and mixed and stirred. Next, the pH was adjusted to 6.4 using citric acid, tri-Na citrate, and ascorbic acid. This aqueous solution was filled into a 100 ml shot bottle, sealed, and subjected to retort sterilization at a temperature of 121 ° C. and 1.2 kgf / cm 2 (using a HIGH-PRESS STEAM STERIZIZER / BS-325 manufactured by Tommy Seiko Co., Ltd.). .
  • Fujioligo G67 is syrup mainly composed of maltohexaose and maltoheptaose.
  • the aggregation state of milk protein in the beverage produced as described above was visually confirmed, and a stability test was performed.
  • a 10% (w / w) aqueous solution of sugar or dextrin used was prepared, and the osmotic pressure was measured with an Osmometer (Fiske Mark 3).
  • the results are shown in Table 27.
  • the protein stability in the table is indicated by the evaluation results of stability ( ⁇ ) and instability ( ⁇ ).
  • the branched megalosaccharide according to the present invention can be easily adjusted to a low osmotic pressure as compared with monosaccharides, disaccharides, and oligosaccharides, and has less burden on the gastrointestinal tract. Moreover, since the milk protein is not destabilized as when dextrin is used, it can be advantageously used for enteral and oral nutrients that require adjustment of osmotic pressure.

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Abstract

Disclosed is a branched megalo-saccharide having aging resistance, flavor-improving effect and the like. Specifically disclosed is a glucan having a polymerization degree of 11-35. The glucan has a branched structure at least at a non-reducing terminal.

Description

新規分岐グルカン並びにその製造方法および用途Novel branched glucan and production method and use thereof 発明の背景Background of the Invention
発明の分野
　本発明は、少なくとも非還元末端に分岐構造を有するグルカンおよびその製造方法に関する。本発明はまた、前記分岐グルカンの用途並びにそれを含有する食品および医薬品に関する。 The present invention relates to a glucan having a branched structure at least at a non-reducing end and a method for producing the glucan. The present invention also relates to uses of the branched glucan and foods and pharmaceuticals containing the same.
背景技術
　デンプンはグルコースを唯一の構成糖とする天然多糖であり、α－１，４－結合からなる主鎖に同様なグルカン鎖が多数α－１，６－結合により結合し、分岐構造が形成された房状構造を有するアミロペクチンと分岐構造を持たない直鎖状のアミロースからなる。デンプンは一般的に水に不溶であるが、熱による変性、あるいは酸や酵素によりある程度加水分解して低分子化することにより水溶性となる。その後、分解を進めることにより、デキストリン、マルトオリゴ糖、ブドウ糖が得られ、これらは一般にデンプン糖と呼ばれる。デンプンの分解度合いはグルコース等量（ＤＥ）により表現される。デキストリンはデンプンを比較的軽度に分解して得られるが、ＤＥが１０以下のものはデキストリン、１０～２０のものはマルトデキストリンと呼称される。 Background Art Starch is a natural polysaccharide composed of glucose as the only saccharide, and a branched structure is formed by binding a number of similar glucan chains to the main chain consisting of α-1,4-linkages through α-1,6-linkages. It is composed of amylopectin having a tufted structure and linear amylose having no branched structure. Starch is generally insoluble in water, but becomes water-soluble by denaturation by heat or hydrolysis to some extent by acid or enzyme to lower the molecular weight. Thereafter, by proceeding with decomposition, dextrin, maltooligosaccharide and glucose are obtained, and these are generally called starch sugar. The degree of starch degradation is expressed in terms of glucose equivalent (DE). Dextrins are obtained by relatively mildly degrading starch, but those with a DE of 10 or less are called dextrins and those with 10-20 are called maltodextrins.
　ＤＥが異なるデンプン分解物はそのＤＥに依存して物性が異なる。すなわち、分解度が高いほど甘味度が高く、最もＤＥが高いブドウ糖において最大となる。一方、ＤＥが低いものほど原料であるデンプン本来の性質である老化性が強く、水溶液中では老化により凝集し、白色沈澱を生じる。また、この老化性は分岐構造が少ないほど顕著であり、分岐構造を持たないアミロースは著しく老化するため、水溶液状態では保存することはできない。また、溶液の粘性についてもＤＥに強く依存する。すなわち、ＤＥが高いものほど水溶液の粘度が低く、ＤＥが低いものほど粘性が高い。このことから、低ＤＥのデキストリンを水分活性が低く、微生物による汚染が問題とならない程度まで濃縮すると溶液の粘度が高すぎるため、操作性が低下する。反対に、操作性に問題ない程度に低粘度な溶液とした場合は、水分活性が高く、微生物汚染により長期保存には適さない。すなわち、デンプンを飲食物の原料として利用する場合、液状品とすることは老化性、粘度、および保存安定性の観点から非現実的であるといえる。このため、デンプンやデキストリンは一般に粉末状の形態で流通している。 The starch degradation products with different DEs have different physical properties depending on the DE. That is, the higher the degree of degradation, the higher the degree of sweetness and the maximum for glucose with the highest DE. On the other hand, the lower the DE, the stronger the aging property, which is the original property of starch as a raw material, and in an aqueous solution, it aggregates due to aging and produces a white precipitate. Further, this aging property becomes more prominent as the branched structure is smaller, and amylose not having a branched structure is significantly aged and cannot be stored in an aqueous solution state. Also, the viscosity of the solution strongly depends on DE. That is, the higher the DE, the lower the viscosity of the aqueous solution, and the lower the DE, the higher the viscosity. For this reason, if the low DE dextrin is concentrated to such an extent that the water activity is low and contamination by microorganisms does not become a problem, the viscosity of the solution is too high and the operability is lowered. On the other hand, when the solution has a viscosity low enough to cause no problem in operability, the water activity is high and it is not suitable for long-term storage due to microbial contamination. That is, when starch is used as a raw material for foods and drinks, it can be said that it is impractical to use a liquid product from the viewpoints of aging, viscosity, and storage stability. For this reason, starch and dextrin are generally distributed in powder form.
　飲食物でデンプン質を利用する場合、デンプンやデキストリンの老化性が品質に影響を与えることが多い。すなわち、飲料では老化による不溶化による白濁、食品では食感の劣化などである。このため、耐老化性に優れたデンプンやデキストリンが求められ、様々な検討がされてきた。耐老化性に優れ、保存安定性が優れたデンプン糖を製造するには老化性を示さない程度、すなわち重合度１０以下までデンプンを分解する、あるいは密な分岐構造を有するようにデンプンを処理する必要があった。例えば、デンプンをβ－アミラーゼにより限界まで分解させた後、生じるマルトースを除去して得られるβ－リミットデキストリンは短鎖の分岐構造を多数有しており、低ＤＥでも耐老化性に優れる特性を有する。しかし、β－リミットデキストリンを工業的に製造するにはマルトースを分画により除去する必要があった。 When starch is used in foods and drinks, the aging properties of starch and dextrin often affect the quality. That is, white turbidity due to insolubilization due to aging in beverages, and deterioration of food texture in foods. For this reason, starch and dextrin excellent in aging resistance have been demanded, and various studies have been made. In order to produce starch sugar having excellent aging resistance and excellent storage stability, starch is decomposed to such an extent that it does not exhibit aging property, that is, the degree of polymerization is 10 or less, or the starch is treated so as to have a dense branched structure. There was a need. For example, β-limit dextrin obtained by degrading starch to the limit with β-amylase and then removing the resulting maltose has many short-chain branched structures, and has excellent aging resistance even at low DE. Have. However, in order to industrially produce β-limit dextrin, it was necessary to remove maltose by fractionation.
　このため、分画などの操作を必要とせず、より簡素にデンプンを改良するためにデンプン液化液に分岐構造を付加する酵素を作用させ、デンプンの非還元末端に分岐構造を付加することにより、耐老化性を高める技術が開発されている（特開２００６－３１２７０５号公報（特許文献１）および特開２００１－２９４６０１号公報（特許文献２））。あるいは、デンプン液化液に、デンプンの分岐鎖を合成する枝作り酵素（ブランチング酵素）、４－α－グルカノトランスフェラーゼまたはシクロデキストリン生成酵素を作用させることにより、水溶性の大環状グルカンを形成させる方法が提案されている (特開平８－１３４１０４号公報（特許文献３）)。さらに、大麦由来の枝作り酵素（ＳＢＥ－ＩＩ）とホスホリラーゼを利用し、グルコース－１－リン酸とマルトオリゴ糖を基質として、グルコース重合度６または７を中心とする分岐構造を有する分岐澱粉を形成させる方法が提案されている (特開２００２－７８４９７号公報（特許文献４）)。 For this reason, an operation for adding a branched structure to the starch liquefaction solution is allowed to act in order to improve starch more simply without requiring operations such as fractionation, and by adding a branched structure to the non-reducing end of starch, Techniques for improving aging resistance have been developed (Japanese Patent Laid-Open No. 2006-312705 (Patent Document 1) and Japanese Patent Laid-Open No. 2001-294601 (Patent Document 2)). Alternatively, a water-soluble macrocyclic glucan is formed by allowing a branching enzyme (branching enzyme), 4-α-glucanotransferase or cyclodextrin-forming enzyme to synthesize starch branch chains to act on the starch liquefaction solution. A method has been proposed (Japanese Patent Laid-Open No. 8-134104 (Patent Document 3)). Furthermore, a branching starch having a branching structure with a glucose polymerization degree of 6 or 7 is formed using glucose-1-phosphate and maltooligosaccharide as substrates using a branching enzyme (SBE-II) derived from barley and phosphorylase. A method of causing this is proposed (Japanese Patent Laid-Open No. 2002-78497 (Patent Document 4)).
　近年、飲料の多様化から味質改善やエネルギー補給などの目的でデキストリン類が添加される場合が多く見受けられる。このような飲料を製造する際、作業性の観点から原料として液状品が求められるケースがある。しかし、上記のようにデンプン質は分子量が大きく、水分活性が微生物の繁殖がない程度に低く、保存安定性に問題ない程度まで濃縮した液糖とした場合は粘度が高すぎるため、操作性が悪いという問題があった。老化に伴う白濁も問題となるが、上記のように耐老化性を高めたデンプンであっても同様に粘度が高すぎるため、液状品の原料として供給することはできない。一方、老化性および操作性が問題とならない程度まで分解度を高めたデンプン糖では、甘味度が高く、茶飲料など低甘味飲料では使用が制限されてしまう。このため、粉末状態だけでなく液状品としても操作性に優れ、また低甘味な糖質の提供が望まれていた。 In recent years, there are many cases where dextrins are added for the purpose of improving taste quality and replenishing energy due to diversification of beverages. When manufacturing such a beverage, there is a case where a liquid product is required as a raw material from the viewpoint of workability. However, as mentioned above, starch has a high molecular weight, water activity is low enough to prevent the growth of microorganisms, and liquid sugar concentrated to such an extent that storage stability does not pose a problem, the viscosity is too high, so the operability is high. There was a problem of being bad. White turbidity associated with aging is also a problem, but even a starch with enhanced aging resistance as described above cannot be supplied as a raw material for a liquid product because the viscosity is also too high. On the other hand, starch sugar having a high degree of degradation to such an extent that aging and operability do not become a problem has a high sweetness, and its use is limited in low-sweet drinks such as tea drinks. Therefore, it has been desired to provide a saccharide that is excellent in operability not only in a powder state but also as a liquid product and has a low sweetness.
　ところで、ポリフェノール類は分子内に複数のフェノール性ヒドロキシル基を持つ植物成分であり、カテキンやアントシアニン、クロロゲン酸などが代表的なポリフェノール類として知られている。ポリフェノール類には、多くの健康増進に有利な生理作用があることが報告されており、例えば、カテキンには抗酸化作用、抗菌作用、脂質代謝改善作用、血圧上昇抑制作用、血糖値上昇抑制作用、消臭作用、抗アレルギー作用などを有する。このような生理活性を利用し、カテキンを高濃度含む緑茶飲料やニアウォーターが健康飲料として広く販売されている。 By the way, polyphenols are plant components having a plurality of phenolic hydroxyl groups in the molecule, and catechins, anthocyanins, chlorogenic acids and the like are known as typical polyphenols. Polyphenols have been reported to have many physiological effects beneficial to health promotion. For example, catechin has an antioxidant action, an antibacterial action, an action to improve lipid metabolism, an action to suppress an increase in blood pressure, and an action to suppress an increase in blood glucose level. , Deodorant action, antiallergic action, etc. Utilizing such physiological activity, green tea beverages and near water containing catechins at high concentrations are widely sold as health beverages.
　しかし、様々な生理機能が知られているがほとんどのポリフェノール類は強い苦味・渋味を呈する。すなわち、ポリフェノール類の強い苦味・渋味のため、高濃度にポリフェノール類を含有せしめた飲食物では、その苦味・渋味により飲食しづらいものとなる。このため、ポリフェノール類には様々な有益な生理機能があるにも関わらず、高濃度摂取することが困難であることが問題である。 However, although various physiological functions are known, most polyphenols exhibit strong bitterness and astringency. That is, due to the strong bitterness and astringency of polyphenols, foods and drinks containing polyphenols at a high concentration are difficult to eat and drink due to the bitterness and astringency. For this reason, although polyphenols have various beneficial physiological functions, it is a problem that it is difficult to ingest high concentrations.
　そこで、ポリフェノール類の味質改善方法として従来から種々の改善策が提案されてきた。例えば、ポリフェノール類の一種であるカテキン類を含有する容器詰飲料に関して、特開２００４－２５４５１１号公報（特許文献５）においてβ－シクロデキストリンを使用する方法が開示されている。しかしながら、β－シクロデキストリンはＡＤＩ値（一日許容摂取量）が５ｍｇ／ｋｇ／日と定められており、使用量が制限されるという課題があり、更にシクロデキストリンを配合することにより飲食物の旨味までも低減してしまうという問題があった。 Therefore, various improvement measures have been conventionally proposed as methods for improving the taste quality of polyphenols. For example, a method of using β-cyclodextrin is disclosed in Japanese Patent Application Laid-Open No. 2004-254511 (Patent Document 5) for a packaged beverage containing catechins which are a kind of polyphenols. However, β-cyclodextrin has an ADI value (daily acceptable intake) of 5 mg / kg / day, and there is a problem that the amount of use is limited. There was a problem that even the taste was reduced.
　特開２００６－２８０２５４号公報（特許文献６）では分岐構造を有する３～４糖を有効成分とする風味調整剤を使用しカテキン類の苦味および／または渋味を抑制する方法が開示されている。しかしながら、糖は低分子であるほど甘味が強く、低分子である３～４糖を多く含有することで甘味の付与が避けられず、緑茶飲料といった低甘味飲食物には使用が制限されていた。 Japanese Patent Application Laid-Open No. 2006-280254 (Patent Document 6) discloses a method for suppressing the bitterness and / or astringency of catechins by using a flavor regulator containing a 3-4 sugar having a branched structure as an active ingredient. . However, the lower the sugar, the stronger the sweetness, and the higher the sugar content of 3-4 sugars, the more sweetness is unavoidable, which limits the use of low-sweet foods such as green tea beverages. .
　また、特開２００８－６１５９３号公報（特許文献７）ではマルトヘキサオースおよび／またはマルトヘプタオースを１．５～６質量％の範囲で配合することによりポリフェノール類の苦味や渋味を低減させる方法が開示されている。しかしながら、この文献で使用されるマルトヘキサオースおよびマルトヘプタオース含有シラップは低分子糖も含有し、甘味を感じやすいために上記と同様な理由で緑茶飲料などの低甘味飲料ではその使用量が制限されるという問題があった。 JP-A-2008-61593 (Patent Document 7) discloses a method for reducing the bitterness and astringency of polyphenols by blending maltohexaose and / or maltoheptaose in a range of 1.5 to 6% by mass. Is disclosed. However, the maltohexaose and maltoheptaose-containing syrups used in this document also contain low-molecular sugars and are easy to feel sweetness, so the amount used is limited in low-sweet drinks such as green tea drinks for the same reason as above. There was a problem of being.
特開２００６－３１２７０５号公報JP 2006-312705 A 特開２００１－２９４６０１号公報JP 2001-294601 A 特開平８－１３４１０４号公報JP-A-8-134104 特開２００２－７８４９７号公報JP 2002-78497 A 特開２００４－２５４５１１号公報JP 2004-254511 A 特開２００６－２８０２５４号公報JP 2006-280254 A 特開２００８－６１５９３号公報JP 2008-61593 A
　本発明者らは、糖転移作用を有する酵素をシクロデキストリン生成酵素と共にデンプン液化液に作用させると、シクロデキストリンをほとんど生成させずに、非還元末端に分岐構造を有する重合度１１～３５程度のグルカンを製造できることを見いだした。本発明者らは、また、非還元末端に分岐構造を有する重合度１１～３５程度のグルカンが、直鎖状マルトデキストリンと比べて極めて高い耐老化性を有するとともに、風味改善や食感の改善等に極めて有効であることを見いだした。本発明はこれらの知見に基づくものである。 The inventors of the present invention, when an enzyme having a sugar transfer action is allowed to act on a starch liquefaction solution together with a cyclodextrin-forming enzyme, hardly produces cyclodextrin and has a degree of polymerization of about 11 to 35 having a branched structure at the non-reducing end. We found that glucan can be produced. The present inventors have also shown that a glucan having a branched structure at the non-reducing end and having a polymerization degree of about 11 to 35 has extremely high aging resistance as compared with a linear maltodextrin, and improves flavor and texture. It was found that it is extremely effective. The present invention is based on these findings.
　すなわち、本発明によれば、以下の発明が提供される。
（１）重合度１１～３５のグルカンであって、少なくとも非還元末端に分岐構造を有するグルカン、またはその還元物（以下「本発明によるメガロ糖」という）。
（２）α－１，４－グルコシド結合により構成されたグルカンと、その非還元末端に結合した分岐構造とからなる、（１）に記載のグルカンまたはその還元物。
（３）分岐構造が、α－１，４－グルコシド結合以外のグルコシド結合により非還元末端に結合した１個以上のグルコース残基により構成される、（２）に記載のグルカンまたはその還元物。
（４）（１）～（３）のいずれか一項に記載のグルカンまたはその還元物を固形分当たり１０～６０質量％含有する液糖または粉糖。
（５）シクロデキストリン生成酵素と糖転移作用を有する酵素とを、デンプン原料に作用させる工程を含んでなる、（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖の製造法。
（６）シクロデキストリン生成酵素と糖転移作用を有する酵素に加えて、枝切り酵素を更に作用させる、（５）に記載の製造法。
（７）シクロデキストリン生成酵素が、パエニバチルス エスピー (Paenibacillus sp.)、バチルス コアギュランス (Bacillus coagulans)、バチルス ステアロサーモフィルス (Bacillus stearothermophilus)、またはバチルス マゼランス (Bacillus macelans) 由来のものである、（５）または（６）に記載の製造法。
（８）糖転移作用を有する酵素が、α－グルコシダーゼ、６－α－グルコシルトランスフェラーゼ、デキストリンデキストラナーゼ、または環状マルトシルマルトース生成酵素である、（５）または（６）に記載の製造法。
（９）α-グルコシダーゼが、アスペルギルス ニガー (Aspergillus niger) またはアクレモニウム エスピー (Acremonium sp.) 由来のものである（８）に記載の製造法。
（１０）枝切り酵素が、イソアミラーゼ、プルラナーゼ、およびこれらの組み合わせからなる群から選択される、（６）に記載の製造法。
（１１）枝切り酵素が、マイロイデス オドラータス (Myroides odoratus)由来イソアミラーゼ、シュードモナス アミロデラモサ (Pseudomonas amyloderamosa) 由来イソアミラーゼ、およびクレブシエラ プネウモニアエ（Klebsiella pneumoniae）由来プルラナーゼ、並びにこれらの組み合わせからなる群から選択される、（６）に記載の製造法。
（１２）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる、食品添加剤または製剤用添加剤。
（１３）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる、食品用風味改善剤または製剤用マスキング剤。
（１４）風味改善が、苦味および／または渋味の低減、酸味の低減、エグ味および／または嫌味の低減、コク味の増強、不快臭の低減、またはアルコール感の低減である、（１３）に記載の風味改善剤。
（１５）風味改善が、苦味および／または渋味の低減である、（１４）に記載の風味改善剤。
（１６）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる、食感改善剤。
（１７）食感改善が、パサパサ感の抑制、しっとり感の付与、またはふんわりした食感の維持である、（１６）に記載の食感改善剤。
（１８）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる、氷の均一的凍結剤。
（１９）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる、食品の照りおよび／またはつや向上剤。
（２０）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を食品に添加することを含んでなる、風味の改善方法または風味が改善された食品の製造方法。
（２１）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を、苦味および／または渋味を有する食品に添加することを含んでなる、苦味および／または渋味の低減化方法または苦味および／または渋味が低減された食品の製造方法。
（２２）苦味および／または渋味を有する食品が、ポリフェノール類含有食品である、（２１）に記載の苦味および／または渋味の低減化方法または苦味および／または渋味が低減された食品の製造方法。
（２３）ポリフェノール類含有食品が、茶系飲料、果実飲料、炭酸飲料、野菜飲料、スポーツ飲料、乳性飲料、アルコール飲料、その他の飲料、アイスクリーム、ゼリー、ムース、飴菓子、ガム、フィリング、健康食品、またはサプリメントである、（２２）に記載の苦味および／または渋味の低減化方法または苦味および／または渋味が低減された食品の製造方法。
（２４）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を、医薬品に添加することを含んでなる、不快な味のマスキング方法または不快な味がマスキングされた医薬品の製造方法。
（２５）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を食品に添加することを含んでなる、食感の改善方法または食感が改善された食品の製造方法。
（２６）添加物が均一に溶解または分散した水に、（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を添加して凍結させることを含んでなる、氷の均一的凍結方法または均一的に凍結した氷の製造方法。
（２７）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を食品に添加することを含んでなる、食品の照りおよび／またはつやの向上方法または照りおよび／またはつやが向上した食品の製造方法。
（２８）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を、乳タンパク質含有飲料に添加することを含んでなる、乳タンパク質の凝集および／または沈殿の抑制方法または乳タンパク質の凝集および／または沈殿が抑制された乳タンパク質含有飲料の製造方法。
（２９）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる、食品。
（３０）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる、ポリフェノール類含有食品。
（３１）ポリフェノール類０．１３質量％に対して、（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物を１．６～５０質量％含有する、（３０）に記載のポリフェノール類含有食品。
（３２）ポリフェノール類含有食品が、茶系飲料、果実飲料、炭酸飲料、野菜飲料、スポーツ飲料、乳性飲料、アルコール飲料、その他の飲料、アイスクリーム、ゼリー、ムース、飴菓子、ガム、フィリング、健康食品、またはサプリメントである、（３１）に記載のポリフェノール類含有食品。
（３３）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる氷、またはそれを含有してなる食品。
（３４）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる、医薬品。
（３５）（１）～（３）のいずれか一項に記載のグルカンもしくはその還元物または（４）に記載の液糖もしくは粉糖を含んでなる、乳タンパク質含有飲料。
（３６）経腸栄養剤または経口栄養剤である、（３５）に記載の乳タンパク質含有飲料。 That is, according to the present invention, the following inventions are provided.
(1) A glucan having a degree of polymerization of 11 to 35 and having a branched structure at least at the non-reducing end, or a reduced product thereof (hereinafter referred to as “megalosaccharide according to the present invention”).
(2) The glucan according to (1) or a reduced product thereof, comprising a glucan composed of α-1,4-glucoside bonds and a branched structure bonded to a non-reducing end thereof.
(3) The glucan or the reduced product thereof according to (2), wherein the branched structure is composed of one or more glucose residues bonded to the non-reducing end by a glucoside bond other than an α-1,4-glucoside bond.
(4) Liquid sugar or powdered sugar containing 10 to 60% by mass of the glucan or reduced product thereof according to any one of (1) to (3) per solid content.
(5) The glucan or reduced product thereof according to any one of (1) to (3), which comprises a step of allowing a cyclodextrin-forming enzyme and an enzyme having a glycosyltransferase action to act on a starch raw material. The method for producing liquid sugar or powdered sugar according to 4).
(6) The production method according to (5), wherein a debranching enzyme is further acted on in addition to the cyclodextrin-forming enzyme and the enzyme having a glycosyltransferase effect.
(7) cyclodextrins synthase, Paenibacillus sp (Paenibacillus sp.), Bacillus coagulans (Bacillus coagulans), Bacillus stearothermophilus (Bacillus stearothermophilus), or Bacillus Mazeransu (Bacillus Macelans) is derived from, (5) Or the manufacturing method as described in (6).
(8) The production method according to (5) or (6), wherein the enzyme having a sugar transfer action is α-glucosidase, 6-α-glucosyltransferase, dextrin dextranase, or cyclic maltosyl maltose producing enzyme.
(9) The production method according to (8), wherein the α-glucosidase is derived from Aspergillus niger or Acremonium sp.
(10) The production method according to (6), wherein the debranching enzyme is selected from the group consisting of isoamylase, pullulanase, and combinations thereof.
(11) The debranching enzyme is selected from the group consisting of an isoamylase derived from Myroides odoratus, an isoamylase derived from Pseudomonas amyloderamosa, a Klebsiella pneumoniae- derived pullulanase, and a combination thereof. The manufacturing method as described in (6).
(12) A food additive or a pharmaceutical additive comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
(13) The glucan according to any one of (1) to (3) or a reduced product thereof, or the flavor improving agent for foods or the masking agent for pharmaceutical preparations comprising the liquid sugar or powdered sugar according to (4) .
(14) The flavor improvement is a reduction in bitterness and / or astringency, a reduction in sourness, a reduction in taste and / or an unpleasant taste, an enhanced richness, a reduction in unpleasant odor, or a reduction in alcoholic feeling. (13) The flavor improving agent as described in 1 ..
(15) The flavor improving agent according to (14), wherein the flavor improvement is a reduction in bitterness and / or astringency.
(16) A texture improving agent comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
(17) The texture-improving agent according to (16), wherein the texture improvement is suppression of a crispy texture, imparting a moist feeling, or maintaining a soft texture.
(18) A homogeneous ice freezing agent comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
(19) A shine and / or gloss improver for food, comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
(20) A method for improving flavor, comprising adding the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4) to food. Or a method for producing a food with improved flavor.
(21) Adding the glucan or reduced product thereof according to any one of (1) to (3) or the liquid sugar or powdered sugar according to (4) to a food having a bitter and / or astringent taste. A method for reducing bitterness and / or astringency, or a method for producing a food with reduced bitterness and / or astringency, comprising:
(22) The method for reducing bitterness and / or astringency according to (21) or the food having reduced bitterness and / or astringency, wherein the food having a bitterness and / or astringency is a polyphenol-containing food. Production method.
(23) Polyphenol-containing foods are tea-based beverages, fruit beverages, carbonated beverages, vegetable beverages, sports beverages, dairy beverages, alcoholic beverages, other beverages, ice creams, jellies, mousses, sweets, gums, fillings, The method for reducing bitterness and / or astringency according to (22) or the method for producing a food with reduced bitterness and / or astringency, which is a health food or a supplement.
(24) An unpleasant taste comprising adding the glucan according to any one of (1) to (3) or a reduced product thereof or the liquid sugar or powdered sugar according to (4) to a pharmaceutical product Masking method or a method for producing a pharmaceutical product with an unpleasant taste masked.
(25) Improvement of texture, comprising adding the glucan or reduced product thereof according to any one of (1) to (3) or the liquid sugar or powdered sugar according to (4) to food. A method or a method for producing a food with improved texture.
(26) Glucan according to any one of (1) to (3) or a reduced product thereof or liquid sugar or powdered sugar according to (4) is added to water in which the additive is uniformly dissolved or dispersed. A method for uniformly freezing ice or a method for producing uniformly frozen ice, which comprises freezing.
(27) Irradiation of food comprising adding the glucan according to any one of (1) to (3) or a reduced product thereof or the liquid sugar or powdered sugar according to (4) to the food, and A method for improving the gloss or / or a method for producing food with improved shine and / or gloss.
(28) adding the glucan or reduced product thereof according to any one of (1) to (3) or the liquid sugar or powdered sugar according to (4) to a milk protein-containing beverage, A method for suppressing aggregation and / or precipitation of milk protein or a method for producing a milk protein-containing beverage in which aggregation and / or precipitation of milk protein is suppressed.
(29) A food comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
(30) A polyphenol-containing food comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
(31) The glucan according to any one of (1) to (3) or the reduced product thereof is contained in an amount of 1.6 to 50% by mass with respect to 0.13% by mass of the polyphenols. Foods containing polyphenols.
(32) Polyphenol-containing foods are tea-based beverages, fruit beverages, carbonated beverages, vegetable beverages, sports beverages, dairy beverages, alcoholic beverages, other beverages, ice creams, jellies, mousses, sweets, gums, fillings, The polyphenol-containing food according to (31), which is a health food or a supplement.
(33) Glucan according to any one of (1) to (3) or a reduced product thereof, or ice containing the liquid sugar or powdered sugar according to (4), or a food containing the same.
(34) A pharmaceutical comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
(35) A milk protein-containing beverage comprising the glucan according to any one of (1) to (3) or a reduced product thereof, or the liquid sugar or powdered sugar according to (4).
(36) The milk protein-containing beverage according to (35), which is an enteral nutrient or an oral nutrient.
　本発明による分岐メガロ糖は、優れた耐老化性を有するとともに、保存安定性や操作性にも優れている。本発明による分岐メガロ糖は、また、不快な味をマスキングするなど風味改善作用を有する。本発明による分岐メガロ糖は更に、糖類などの混合成分を含有する水に添加した場合に氷の均一性を向上・促進させる作用を有する。本発明による分岐メガロ糖はまた、食品の照りやつやを向上させる作用を有する。本発明による分岐メガロ糖は、更にまた、低甘味であるとともに、食品に添加しても食品本来の風味に影響を与えない。本発明による分岐メガロ糖は、また、乳タンパク質の凝集や沈殿を防止し、乳タンパク質を安定して存在させることができる。従って、本発明による分岐メガロ糖およびその還元物並びにそれを含有する液糖および粉糖は、食品添加物や製剤用添加剤として幅広く実用可能である。 The branched megalosaccharide according to the present invention has excellent aging resistance, as well as excellent storage stability and operability. The branched megalosaccharide according to the present invention also has a taste improving action such as masking unpleasant taste. The branched megalosaccharide according to the present invention further has an action of improving and promoting ice uniformity when added to water containing a mixed component such as saccharide. The branched megalosaccharide according to the present invention also has an effect of improving the shine and gloss of food. Furthermore, the branched megalosaccharide according to the present invention has a low sweetness and does not affect the original flavor of food even when added to food. The branched megalosaccharide according to the present invention can also prevent milk protein aggregation and precipitation and allow milk protein to exist stably. Therefore, the branched megalosaccharide and its reduced product according to the present invention, and liquid sugar and powdered sugar containing the same can be widely used as food additives and pharmaceutical additives.
　本発明による製造方法によれば、本発明による分岐メガロ糖を簡便かつ高収率で製造できる。従って、本発明による製造方法は、本発明による分岐メガロ糖の工業生産に途を拓くものである。 According to the production method of the present invention, the branched megalosaccharide according to the present invention can be produced simply and with high yield. Therefore, the production method according to the present invention opens up the industrial production of branched megalosaccharide according to the present invention.
本発明による分岐メガロ糖の耐老化性を示した図である。１：α－グルコシダーゼ未添加区、２：α－グルコシダーゼ添加区。It is the figure which showed the aging resistance of the branched megalo sugar by this invention. 1: no α-glucosidase added group, 2: α-glucosidase added group. 本発明による分岐メガロ糖の耐老化性をコーンシラップとの対比で示した図である。１：分岐メガロ糖含有シラップ、２：コーンシラップ。It is the figure which showed the aging resistance of the branched megalo sugar by this invention by contrast with corn syrup. 1: Branched megalosaccharide-containing syrup, 2: corn syrup.
発明の具体的説明DETAILED DESCRIPTION OF THE INVENTION
分岐メガロ糖およびその製造
　本発明による分岐メガロ糖は、直鎖状グルカンと分岐構造とからなる重合度１１～３５のグルカンであって、少なくとも直鎖状グルカンの非還元末端に分岐構造が導入されたグルカンである。ここで、「直鎖状グルカン」とは、単一のグルコシド結合によりグルコース分子が結合して構成された直鎖状のグルカンを意味する。 Branched megalosaccharide and production thereof The branched megalosaccharide according to the present invention is a glucan having a polymerization degree of 11 to 35 composed of a linear glucan and a branched structure, and a branched structure is introduced at least at the non-reducing end of the linear glucan. It is a glucan. Here, the “linear glucan” means a linear glucan composed of glucose molecules bonded by a single glucoside bond.
　本発明による分岐メガロ糖の具体例としては、α－１，４－グルコシド結合により構成された直鎖状グルカンと、少なくともその直鎖状グルカンの非還元末端に導入された分岐構造とからなる構造を有する、重合度１１～３５の分岐メガロ糖が挙げられる。本発明による分岐メガロ糖の更なる具体例としては、α－１，４－グルコシド結合により構成された直鎖状グルカンと、その直鎖状グルカンの非還元末端のみに導入された分岐構造とからなる構造を有する、重合度１１～３５の分岐メガロ糖が挙げられる。 Specific examples of the branched megalosaccharide according to the present invention include a structure comprising a linear glucan composed of α-1,4-glucoside bonds and a branched structure introduced at least at the non-reducing end of the linear glucan. And branched megalosaccharide having a polymerization degree of 11 to 35. Further specific examples of the branched megalosaccharide according to the present invention include a linear glucan constituted by α-1,4-glucoside bonds and a branched structure introduced only at the non-reducing end of the linear glucan. And a branched megalosaccharide having a polymerization degree of 11 to 35.
　本発明において「分岐構造」とは、α－１，４－グルコシド結合以外のグルコシド結合により直鎖状グルカンに結合した１個以上のグルコース残基からなるグルカン残基を意味する。α－１，４－グルコシド結合以外のグルコシド結合としては、α－１，６－グルコシド結合、α－１，３－グルコシド結合、α－１，２－グルコシド結合が挙げられる。 In the present invention, the “branched structure” means a glucan residue composed of one or more glucose residues bonded to a linear glucan by a glucoside bond other than an α-1,4-glucoside bond. Examples of glucoside bonds other than α-1,4-glucoside bonds include α-1,6-glucoside bonds, α-1,3-glucoside bonds, and α-1,2-glucoside bonds.
　後述するように、本発明による製造方法で使用される糖転移作用を有する酵素を選択することによって、非還元末端に導入される分岐構造を変化させることができる。分岐構造のグルカン残基を構成するグルコース残基の個数は本発明による分岐メガロ糖の重合度を満たす限り特に限定されないが、好ましくは、１～数個、より好ましくは、１～６個、１～４個、１～３個、または１～２個とすることができる。 As will be described later, the branch structure introduced into the non-reducing end can be changed by selecting an enzyme having a glycosyltransferase effect used in the production method according to the present invention. The number of glucose residues constituting the branched glucan residue is not particularly limited as long as the degree of polymerization of the branched megalosaccharide according to the present invention is satisfied, but preferably 1 to several, more preferably 1 to 6, 1 There can be ˜4, 1-3, or 1-2.
　分岐構造が２糖単位で構成されたグルカン残基の場合には、その分岐構造としてはコージビオース、ニゲロース、マルトース、イソマルトース構造が挙げられ、より具体的には、直鎖状グルカンの非還元末端にα－１，４－結合以外の結合様式でコージビオース、ニゲロース、マルトース、またはイソマルトースが結合した構造が挙げられる。分岐構造が３糖以上のグルカン残基の場合には、その分岐構造としては、イソマルトトリオース、ニゲロトリオースなど単一なグルコシド結合のみから構成されるグルカンや、パノースなど複数のグルコシド結合により構成されたグルカンが挙げられ、より具体的には、直鎖状グルカンの非還元末端にα－１，４－結合以外の結合様式で、イソマルトトリオース、ニゲロトリオースなど単一なグルコシド結合のみから構成されるグルカンや、パノースなど複数のグルコシド結合により構成されたグルカンが結合した構造が挙げられる。 In the case where the branched structure is a glucan residue composed of disaccharide units, examples of the branched structure include cordobiose, nigerose, maltose, isomaltose structure, and more specifically, the non-reducing end of a linear glucan. In addition, a structure in which cordobiose, nigerose, maltose, or isomaltose is bonded in a bonding mode other than α-1,4-bond. In the case where the branched structure is a glucan residue having 3 or more sugars, the branched structure may be a glucan composed only of a single glucoside bond such as isomaltotriose or nigerotriose, or a plurality of glucoside bonds such as panose. More specifically, a single glucoside bond such as isomaltotriose or nigerotriose is used at the non-reducing end of a linear glucan with a binding mode other than α-1,4-linkage. And a structure in which a glucan composed of a plurality of glucoside bonds such as panose is bound.
　本発明において「還元末端」とは、還元性を示す糖残基を意味する。本発明において「非還元末端」とは、還元性を示さない糖残基、すなわち、「還元末端」以外の末端糖残基を意味する。 In the present invention, the “reducing terminal” means a sugar residue that exhibits reducing properties. In the present invention, the “non-reducing terminal” means a sugar residue that does not exhibit reducing properties, that is, a terminal sugar residue other than the “reducing terminal”.
　本発明において「重合度」とは、グルカンを構成するグルコース残基の個数を指し、直鎖状グルカンを構成するグルコース残基の個数のみならず、分岐構造を構成するグルコース残基の個数を含む。分岐糖類の重合度は、高速液体クロマトグラフィー (ＨＰＬＣ)法によって測定することができる。 In the present invention, the “degree of polymerization” refers to the number of glucose residues constituting a glucan and includes not only the number of glucose residues constituting a linear glucan but also the number of glucose residues constituting a branched structure. . The degree of polymerization of the branched saccharide can be measured by high performance liquid chromatography (HPLC) method.
　本発明によるメガロ糖の重合度は１１～３５であるが、重合度を１５～３５の範囲とすることもできる。なお、本明細書では、重合度１１～３５のグルカンを単に「メガロ糖」ということがある。 The degree of polymerization of megalosaccharide according to the present invention is 11 to 35, but the degree of polymerization may be in the range of 15 to 35. In the present specification, a glucan having a degree of polymerization of 11 to 35 is sometimes simply referred to as “megalosugar”.
　本発明において「還元物」とは、糖の還元末端のグルコシル基のアルデヒド基が還元され、水酸基となっているものを言う。 In the present invention, “reduced product” refers to a product in which the aldehyde group of the glucosyl group at the reducing end of the sugar is reduced to form a hydroxyl group.
　糖の還元物を得る方法は当業者に周知であり、使用可能な還元方法を例示すれば、ヒドリド還元剤を用いる方法、プロトン性溶媒中の金属を用いる方法、電解還元方法、接触水素化反応方法等が挙げられる。本発明においては、少量の還元物を調製する場合にはヒドリド還元剤を用いる方法が簡便且つ特殊な装置を必要とせず便利であり、一方で、工業的に大規模に実施する場合には、経済性優れ、副生成物も少ないという点から、接触水素化反応を用いる方法が好ましい。 Methods for obtaining a reduced product of sugar are well known to those skilled in the art. Examples of usable reduction methods include a method using a hydride reducing agent, a method using a metal in a protic solvent, an electrolytic reduction method, and a catalytic hydrogenation reaction. Methods and the like. In the present invention, when a small amount of reductant is prepared, the method using a hydride reducing agent is convenient and convenient without requiring a special apparatus, while on the other hand, when carried out on an industrial scale, A method using a catalytic hydrogenation reaction is preferred because it is economical and has few by-products.
　接触水素化反応とは、触媒の存在下、不飽和有機化合物の二重結合部に水素を添加する反応であり、一般に水添反応とも言われている。本発明による還元物の製造方法を具体的に説明すると、本発明による分岐メガロ糖を水に溶解し、そこにラネーニッケル触媒を適量加え、水素ガスを添加し、高温条件下で還元する。次に、脱色・脱イオン処理して、分岐メガロ糖還元糖組成物を得る。 The catalytic hydrogenation reaction is a reaction in which hydrogen is added to the double bond portion of an unsaturated organic compound in the presence of a catalyst, and is generally also referred to as a hydrogenation reaction. The method for producing the reduced product according to the present invention will be specifically described. The branched megalosaccharide according to the present invention is dissolved in water, an appropriate amount of Raney nickel catalyst is added thereto, hydrogen gas is added, and reduction is performed under high temperature conditions. Next, decolorization / deionization treatment is performed to obtain a branched megalosaccharide reducing sugar composition.
　接触水素化反応において使用し得る触媒としては、公知の水添触媒なら特に限定されないが、例えば、ラネーニッケル、還元ニッケル、珪藻土、アルミナ、軽石、シリカゲル、酸性白土などの種々の担体に担持したニッケル－担体触媒などのニッケル触媒；ラネーコバルト、還元コバルト、コバルト－担体触媒などのコバルト触媒；ラネー銅、還元銅、銅－担体触媒などの銅触媒；パラジウム黒、酸化パラジウム、コロイドパラジウム、パラジウム－炭素、パラジウム－硫酸バリウム、パラジウム－酸化マグネシウム、パラジウム－アルミナなどのパラジウム触媒；白金黒、コロイド白金、酸化白金、硫化白金、白金－炭素などの白金－担体触媒等の白金触媒；コロイドロジウム、ロジウム－炭素、酸化ロジウムなどのロジウム触媒；ルテニウム触媒などの白金族触媒；酸化二レニウム、レニウム－炭素などのレニウム触媒；銅クロム酸化物触媒；三酸化モリブデン触媒；酸化バナジウム触媒；酸化タングステン触媒；銀触媒などが挙げられる。これらの触媒の内では、ラネーニッケル、還元ニッケル、ニッケル珪藻土を用いることが好ましく、より好ましくは、ラネーニッケルである。 The catalyst that can be used in the catalytic hydrogenation reaction is not particularly limited as long as it is a known hydrogenation catalyst. For example, nickel-supported on various supports such as Raney nickel, reduced nickel, diatomaceous earth, alumina, pumice, silica gel, and acid clay. Nickel catalysts such as supported catalysts; cobalt catalysts such as Raney cobalt, reduced cobalt, cobalt-supported catalysts; copper catalysts such as Raney copper, reduced copper, copper-supported catalysts; palladium black, palladium oxide, colloidal palladium, palladium-carbon, Palladium catalysts such as palladium-barium sulfate, palladium-magnesium oxide, palladium-alumina; platinum catalysts such as platinum-support catalysts such as platinum black, colloidal platinum, platinum oxide, platinum sulfide, platinum-carbon; colloidal rhodium, rhodium-carbon Rhodium catalysts such as rhodium oxide; Platinum group catalysts such as um catalyst; -; copper chromium oxide catalyst; molybdenum trioxide catalyst; vanadium oxide catalyst; tungsten oxide catalyst; dinitrogen rhenium, rhenium rhenium catalyst such as carbon such as silver catalyst. Among these catalysts, it is preferable to use Raney nickel, reduced nickel, or nickel diatomaceous earth, and more preferably Raney nickel.
　また、水素の圧力は通常１０～２５０ｋｇ／ｃｍ２、好ましく５０～２００ｋｇ／ｃｍ２の範囲である。また、反応温度は触媒量、溶媒種別により異なるが、通常８０～２００℃の範囲であることが好ましく、９０～１６０℃がより好ましい。 The hydrogen pressure is usually in the range of 10 to 250 kg / cm 2 , preferably 50 to 200 kg / cm 2 . The reaction temperature varies depending on the amount of catalyst and the type of solvent, but is usually preferably in the range of 80 to 200 ° C, more preferably 90 to 160 ° C.
　なお、本明細書において「本発明による分岐メガロ糖」というときは、還元物をも含む意味で用いられるものとする。 In the present specification, the term “branched megalosaccharide according to the present invention” is intended to include a reduced product.
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、シクロデキストリン生成酵素および糖転移作用を有する酵素を、デンプン原料に作用させることにより製造することができる。 The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can be produced by allowing a cyclodextrin-forming enzyme and an enzyme having a sugar transfer action to act on a starch raw material.
　本発明による製造方法では、デンプン原料に、シクロデキストリン生成酵素および糖転移作用を有する酵素を一緒に作用させることが好ましい。 In the production method according to the present invention, it is preferable to cause the starch raw material to act together with a cyclodextrin-producing enzyme and an enzyme having a sugar transfer action.
　以下に拘束される訳ではないが、分岐メガロ糖の生成機構は次のようなものであると考えられる。すなわち、デンプン原料に含まれるデキストリンの非還元末端、あるいはシクロデキストリン生成酵素の加水分解、カップリング、不均化反応のいずれかにおいて低分子化されたデキストリンの非還元性末端にα－グルコシダーゼが作用してα－１，４－結合を切断し、グルコシル基を他のあるいは同一の非還元性末端のグルコシル基にα－１，６－結合、α－１，２－結合、あるいはα－１，３－結合で付加する。これにより非還元性末端に分岐構造を有するメガロ糖が生じる。反応初期はこのような分岐メガロ糖が反応系内に存在しないため、シクロデキストリン生成酵素は反応初期にはシクロデキストリンを生じる。しかし、反応後期では大半のマルトデキストリンの非還元性末端に分岐鎖が付加されるため、このような分岐構造を有する糖質はシクロデキストリン生成酵素の環状化反応の基質とはならない。このため、シクロデキストリン生成酵素によるシクロデキストリン生成反応は反応初期にしか起こらず、また、シクロデキストリン生成酵素のカップリング反応により生じたシクロデキストリンが開環され、α－グルコシダーゼによる糖転移反応の基質として供給される。その結果、反応初期に生じたシクロデキストリンは反応後期にはほぼ完全に分解し、反応後期にはシクロデキストリンはほとんど残存しない。枝切り酵素を反応液中に共存させた場合には、デンプン分岐鎖を切断し、直鎖状のデキストリンを供給するため、シクロデキストリン生成酵素によるデンプンの低分子化を促進する他、このような直鎖状のデキストリンはカップリング反応における受容体分子としても働くため、反応を効率的に進めることが可能となると考えられる。 Although not limited to the following, it is considered that the production mechanism of branched megalosaccharide is as follows. That is, α-glucosidase acts on the non-reducing end of dextrin contained in starch raw material, or on the non-reducing end of dextrin that has been reduced in molecular weight in any of hydrolysis, coupling, or disproportionation of cyclodextrin-forming enzyme. Then, the α-1,4-bond is cleaved, and the glucosyl group is linked to another or the same non-reducing terminal glucosyl group by α-1,6-bond, α-1,2-bond, or α-1, 3- Add with a bond. This produces a megalosaccharide having a branched structure at the non-reducing end. Since such a branched megalosaccharide does not exist in the reaction system at the beginning of the reaction, the cyclodextrin-producing enzyme produces cyclodextrin at the beginning of the reaction. However, since a branched chain is added to the non-reducing end of most maltodextrins in the late stage of the reaction, a saccharide having such a branched structure does not serve as a substrate for the cyclization reaction of cyclodextrin-producing enzyme. For this reason, the cyclodextrin-producing reaction by the cyclodextrin-producing enzyme occurs only in the early stage of the reaction, and the cyclodextrin produced by the coupling reaction of the cyclodextrin-producing enzyme is opened and used as a substrate for the transglycosylation reaction by α-glucosidase. Supplied. As a result, cyclodextrin produced in the early stage of the reaction is almost completely decomposed in the later stage of the reaction, and almost no cyclodextrin remains in the later stage of the reaction. When a debranching enzyme coexists in the reaction solution, the starch branch chain is cleaved and linear dextrin is supplied. Since linear dextrin also functions as an acceptor molecule in the coupling reaction, it is considered that the reaction can proceed efficiently.
　デンプン原料から分岐メガロ糖を生成させる酵素反応は、酵素反応が進行する温度で実施することができ、通常、６０℃付近までの温度範囲で実施することができる。好適な反応温度は、３０～５５℃である。デンプン原料から分岐メガロ糖を生成させる酵素反応は、酵素反応が進行するｐＨで実施することができ、通常、ｐＨ５～９の範囲で実施することができる。好適な反応ｐＨは、ｐＨ５．５～７の範囲である。酵素の使用量と反応時間とは密接に関係しており、目的とする酵素反応の進行により適宜反応時間を調節することができる。 The enzymatic reaction for producing branched megalosaccharide from starch raw material can be carried out at a temperature at which the enzymatic reaction proceeds, and can usually be carried out in a temperature range up to around 60 ° C. The preferred reaction temperature is 30-55 ° C. The enzymatic reaction for producing branched megalosaccharide from the starch raw material can be carried out at a pH at which the enzymatic reaction proceeds, and can usually be carried out in the range of pH 5-9. A suitable reaction pH is in the range of pH 5.5-7. The amount of enzyme used and the reaction time are closely related, and the reaction time can be appropriately adjusted according to the progress of the target enzyme reaction.
　本発明における製造方法ではデンプン原料としてデンプン液化液を用いることができる。原料として用いられるデンプン液化液のデンプン濃度は、酵素反応の効率やデンプンの溶解度等の観点から、１０～４５質量％とすることができる。但し、デンプン部分分解物を原料とする場合は基質濃度が４５質量％を越えても反応を良好に進行させることができる。デンプン液化液のＤＥは、通常ＤＥ２～２５の範囲とすることが好ましく、より好ましくはＤＥ３～１０の範囲である。本発明による製造方法での酵素反応に際しては、オリゴ糖の遊離を防ぐため、液化酵素を失活させることが好ましい。本発明における製造方法では、デンプン原料としてデンプン糊化物を用いてもよい。 In the production method of the present invention, a starch liquefied liquid can be used as a starch raw material. The starch concentration of the starch liquefaction solution used as a raw material can be set to 10 to 45% by mass from the viewpoint of the efficiency of enzyme reaction, the solubility of starch, and the like. However, when the starch partial decomposition product is used as a raw material, the reaction can proceed well even if the substrate concentration exceeds 45% by mass. The DE of the starch liquefaction liquid is usually preferably in the range of DE 2 to 25, more preferably in the range of DE 3 to 10. In the enzyme reaction in the production method according to the present invention, it is preferable to deactivate the liquefying enzyme in order to prevent the oligosaccharide from being released. In the production method of the present invention, starch gelatinized material may be used as a starch raw material.
　本発明による製造方法で酵素反応に用いられるシクロデキストリン生成酵素の添加量は、反応効率および製造コストの観点から、対基質（固形）１ｇ当たり０．２～１０単位とすることができる。ここで、シクロデキストリン生成酵素１単位とは後述するシクロデキストリン生成酵素の活性測定方法の条件下において、１分間に１ｍｇのβ－シクロデキストリンを生成するのに必要な酵素量をいう。 The amount of cyclodextrin-producing enzyme used in the enzyme reaction in the production method according to the present invention can be 0.2 to 10 units per gram of substrate (solid) from the viewpoint of reaction efficiency and production cost. Here, 1 unit of cyclodextrin-producing enzyme means the amount of enzyme necessary to produce 1 mg of β-cyclodextrin per minute under the conditions of the method for measuring the activity of cyclodextrin-producing enzyme described later.
　本発明による製造方法で酵素反応に用いられる糖転移作用を有する酵素のうちα－グルコシダーゼの添加量は、反応効率および製造コストの観点から、対基質（固形）１ｇ当たり０．０１～３０単位とすることができる。ここで、α－グルコシダーゼ１単位とは後述するα－グルコシダーゼの活性測定方法の条件下において、１分間に１μｍｏｌのマルトースを加水分解するのに必要な酵素量をいう。 Among the enzymes having a glycosyltransferase effect used in the enzyme reaction in the production method according to the present invention, the amount of α-glucosidase added is 0.01 to 30 units per gram of substrate (solid) from the viewpoint of reaction efficiency and production cost. can do. Here, one unit of α-glucosidase refers to the amount of enzyme required to hydrolyze 1 μmol of maltose per minute under the conditions of the α-glucosidase activity measurement method described later.
　α－グルコシダーゼ以外の糖転移作用を有する酵素の添加量については、酵素反応や反応条件は周知であることから、当業者であれば、α－グルコシダーゼの添加量に従ってその添加量を決定できる。 Since the enzyme reaction and reaction conditions for the addition amount of an enzyme having a glycosyltransferase other than α-glucosidase are well known, those skilled in the art can determine the addition amount according to the addition amount of α-glucosidase.
　本発明による製造方法では、シクロデキストリン生成酵素と糖転移作用を有する酵素に加えて、枝切り酵素を更に作用させることができる。枝切り酵素を作用させた場合は、分岐メガロ糖生成反応後に残存する重合度３５を超えるデキストリン成分の残存量が少なく、分岐メガロ糖生成量が高まることから、効率的に分岐メガロ糖を製造するためには枝切り酵素を使用することが好ましい。 In the production method according to the present invention, a debranching enzyme can be further acted in addition to the cyclodextrin-forming enzyme and the enzyme having a sugar transfer action. When a debranching enzyme is allowed to act, since the residual amount of dextrin components exceeding the degree of polymerization of 35 remaining after the branching megalosaccharide production reaction is small and the amount of branched megalosaccharide production is increased, the branched megalosaccharide is efficiently produced. For this purpose, it is preferable to use a debranching enzyme.
　枝切り酵素は、シクロデキストリン生成酵素および糖転移作用を有する酵素と一緒に、デンプン原料に作用させることが好ましい。 The debranching enzyme is preferably allowed to act on the starch raw material together with the cyclodextrin-forming enzyme and the enzyme having a sugar transfer action.
　本発明による製造方法で酵素反応に用いられる枝切り酵素のうちイソアミラーゼの添加量は、反応効率および製造コストの観点から、対基質（固形）１ｇ当たり１０～１０００単位とすることができる。本発明による製造方法で酵素反応に用いられる枝切り酵素のうちプルラナーゼの添加量は、反応性および製造コストの観点から、対基質（固形）当たり０．００１～０．１質量％とすることができる。ここで、イソアミラーゼ１単位とは、後述するイソアミラーゼの活性測定方法の条件下において６１０ｎｍの吸光度を０．０１増加させる酵素力価である。 In the production method according to the present invention, among the debranching enzymes used in the enzymatic reaction, the amount of isoamylase added can be 10 to 1000 units per gram of substrate (solid) from the viewpoint of reaction efficiency and production cost. Of the debranching enzyme used in the enzymatic reaction in the production method according to the present invention, the amount of pullulanase added may be 0.001 to 0.1% by mass per substrate (solid) from the viewpoint of reactivity and production cost. it can. Here, 1 unit of isoamylase is an enzyme titer that increases the absorbance at 610 nm by 0.01 under the conditions of the method for measuring the activity of isoamylase described later.
　イソアミラーゼおよびプルラナーゼ以外の枝切り酵素の添加量については、当業者であれば、イソアミラーゼやプルラナーゼの添加量に従ってその添加量を決定できる。 Regarding the addition amount of the debranching enzyme other than isoamylase and pullulanase, those skilled in the art can determine the addition amount according to the addition amount of isoamylase or pullulanase.
　本発明による製造方法の原料となるデンプンの由来は特に限定されないが、例えば、コーンスターチ、ワキシーコーンスターチ、米デンプン、餅米デンプン、小麦デンプン、サゴヤシデンプンなどの地上デンプンや、馬鈴薯デンプン、甘藷デンプン、タピオカ澱デンプン、くずデンプンなどの地下デンプンを用いることができる。さらに、デンプンから得られたアミロース、アミロペクチン、デンプン部分分解物などを原料とすることも可能である。これらのデンプンは液化あるいは糊化して本発明による製造方法に使用することができる。例えば、デンプンにアミラーゼなどの液化酵素を作用させて得られたデンプン液化液を本発明による製造方法のデンプン原料として用いることができる。デンプンの液化の方法や糊化の方法は当業者に周知であり、いずれの方法をも用いることができる。 The origin of the starch used as the raw material for the production method according to the present invention is not particularly limited. Subsurface starches such as starch starch and waste starch can be used. Furthermore, it is also possible to use amylose, amylopectin, starch partial decomposition product, etc. obtained from starch as a raw material. These starches can be liquefied or gelatinized and used in the production method according to the present invention. For example, a starch liquefied liquid obtained by allowing a liquefying enzyme such as amylase to act on starch can be used as a starch raw material in the production method according to the present invention. Methods for liquefying starch and gelatinizing are well known to those skilled in the art, and any method can be used.
　本発明による製造方法に用いる酵素は、分岐メガロ糖を調製できる限り、精製酵素であっても粗酵素であっても良く、また、遊離の酵素であっても、固定化された酵素であってもよい。固定化酵素の場合、反応の形式は、バッチ式、半連続式および連続式のいずれでもよい。固定化方法としては、担体結合法、（例えば、共有結合法、イオン結合法、あるいは物理的吸着法）、架橋法あるいは包括法（格子型あるいはマイクロカプセル型）など、公知の方法を使用することができる。 The enzyme used in the production method according to the present invention may be a purified enzyme or a crude enzyme as long as a branched megalosaccharide can be prepared, and it may be a free enzyme or an immobilized enzyme. Also good. In the case of an immobilized enzyme, the reaction format may be any of batch, semi-continuous and continuous. As the immobilization method, a known method such as a carrier bonding method (for example, a covalent bonding method, an ionic bonding method, or a physical adsorption method), a crosslinking method, or a comprehensive method (lattice type or microcapsule type) should be used. Can do.
　本発明による製造方法に用いる「シクロデキストリン生成酵素」は、市販のものを用いても、微生物から単離したものを用いてもよい。単離源となる微生物は、天然由来の微生物に加えて、シクロデキストリン生成酵素産生能を有する組換え微生物や、天然由来の微生物を変異させた変異株であってもよい。「シクロデキストリン生成酵素」の微生物起源は特に限定されないが、例えば、パエニバチルス エスピー (Paenibacillus sp.)、バチルス コアギュランス (Bacillus coagulans)、バチルス ステアロサーモフィルス (Bacillus stearothermophilus)、およびバチルス マゼランス (Bacillus macelans) 由来のものを用いることができる。 As the “cyclodextrin-producing enzyme” used in the production method according to the present invention, a commercially available product or a product isolated from a microorganism may be used. The microorganism as an isolation source may be a recombinant microorganism having the ability to produce a cyclodextrin-producing enzyme, or a mutant strain obtained by mutating a naturally-occurring microorganism, in addition to a naturally-occurring microorganism. Microbial origin of "cyclodextrin forming enzyme" is not particularly limited, for example, Paenibacillus sp (Paenibacillus sp.), Bacillus coagulans (Bacillus coagulans), Bacillus stearothermophilus (Bacillus stearothermophilus), and Bacillus Mazeransu (Bacillus macelans) from Can be used.
　本発明による製造方法に用いる「糖転移作用を有する酵素」としては、α－グルコシダーゼ、６－α－グルコシルトランスフェラーゼ、デキストリンデキストラナーゼ、および環状マルトシルマルトース生成酵素が挙げられる。 Examples of the “enzyme having glycosyltransferase effect” used in the production method according to the present invention include α-glucosidase, 6-α-glucosyltransferase, dextrin dextranase, and cyclic maltosyl maltose producing enzyme.
　α-グルコシダーゼは、市販のものを用いても、微生物から単離したものを用いてもよい。単離源となる微生物は、天然由来の微生物に加えて、α－グルコシダーゼ生成酵素産生能を有する組換え微生物や、天然由来の微生物を変異させた変異株であってもよい。α－グルコシダーゼの微生物起源は特に限定されないが、例えば、アスペルギルス ニガー (Aspergillus niger) およびアクレモニウム エスピー (Acremonium sp.) 由来のものを用いることができる。 As α-glucosidase, a commercially available product or a product isolated from a microorganism may be used. The microorganism as an isolation source may be a recombinant microorganism having the ability to produce an α-glucosidase-producing enzyme, or a mutant strain obtained by mutating a naturally-occurring microorganism, in addition to a naturally-occurring microorganism. The microbial origin of α-glucosidase is not particularly limited. For example, those derived from Aspergillus niger and Acremonium sp. can be used.
　本発明による製造方法に用いる「枝切り酵素」としては、イソアミラーゼ、プルラナーゼが挙げられ、長鎖な分岐鎖を切断しやすいイソアミラーゼを使用することが好ましい。 Examples of the “branching enzyme” used in the production method according to the present invention include isoamylase and pullulanase, and it is preferable to use an isoamylase that can easily cleave a long branched chain.
　イソアミラーゼは、市販のものを用いても、微生物から単離したものを用いてもよい。
　単離源となる微生物は、天然由来の微生物に加えて、イソアミラーゼ産生能を有する組換え微生物や、天然由来の微生物を変異させた変異株であってもよい。イソアミラーゼの微生物起源は特に限定されないが、例えば、マイロイデス オドラータス (Myroides odoratus)およびシュードモナス アミロデラモサ (Pseudomonas amyloderamosa)由来のものを用いることができる。 As the isoamylase, a commercially available product or a product isolated from a microorganism may be used.
The microorganism as an isolation source may be a recombinant microorganism having isoamylase-producing ability or a mutant strain obtained by mutating a naturally-occurring microorganism in addition to a naturally-occurring microorganism. Although microbial origin isoamylase is not particularly limited, for example, Mairoidesu Odoratasu (Myroides odoratus) and Pseudomonas Amiroderamosa (Pseudomonas amyloderamosa) can be used those derived from.
　プルラナーゼは、市販のものを用いても、微生物から単離したものを用いてもよい。単離源となる微生物は、天然由来の微生物に加えて、プルラナーゼ産生能を有する組換え微生物や、天然由来の微生物を変異させた変異株であってもよい。プルラナーゼの微生物起源は特に限定されないが、例えば、クレブシエラ プネウモニアエ (Klebseilla pneumoniae) やバチルス ブレビス (Bacillus brevis)由来のものを用いることができる。プルラナーゼとしては市販酵素を利用する場合には、好ましくは、天野エンザイム社製プルラナーゼ「アマノ」３を使用することができる。 As the pullulanase, a commercially available product or a product isolated from a microorganism may be used. The microorganism as an isolation source may be a recombinant microorganism having pullulanase-producing ability or a mutant strain obtained by mutating a naturally-occurring microorganism in addition to a naturally-occurring microorganism. Although microbial origin of pullulanase is not particularly limited, for example, Klebsiella pneumoniae (Klebseilla pneumoniae) and brevis (Bacillus brevis) can be used those derived from. When a commercially available enzyme is used as the pullulanase, pullulanase “Amano” 3 manufactured by Amano Enzyme can be preferably used.
　本発明による製造方法を実施すると、反応物中に本発明による分岐メガロ糖を得ることができる。反応物中に存在するメガロ糖の非還元末端に分岐構造が導入されているかは、マルトデキストリンやデンプンの非還元性末端から２糖単位でα－１，４－結合を加水分解するβ－アミラーゼにより加水分解を受けないことにより確認することができる。使用できるβ－アミラーゼは、特に限定されないが、例えば、大豆由来のβ－アミラーゼを使用することができる。なお、β－アミラーゼと同様にマルトデキストリンやデンプンの非還元性末端よりα－１，４－結合を加水分解するグルコアミラーゼを使用することも可能ではあるが、この酵素はα－１，６－結合分解能も有するため、β－アミラーゼを使用することが好ましい。 When the production method according to the present invention is carried out, the branched megalosaccharide according to the present invention can be obtained in the reaction product. Whether a branched structure is introduced at the non-reducing end of the megalo sugar present in the reaction product is determined by β-amylase that hydrolyzes α-1,4-bonds in units of two sugars from the non-reducing end of maltodextrin or starch Can be confirmed by not undergoing hydrolysis. The β-amylase that can be used is not particularly limited. For example, soybean-derived β-amylase can be used. As with β-amylase, it is possible to use maltodextrin or glucoamylase that hydrolyzes α-1,4-linkage from the non-reducing end of starch, but this enzyme is α-1,6- Since it also has a binding resolution, it is preferable to use β-amylase.
　本発明による製造方法では、糖転移酵素としてアクレモニウム　エスピー由来のα－グルコシダーゼを使用すると、グルコース残基がα－１，３－グルコシド結合により非還元末端に結合した分岐メガロ糖を製造することができる。この場合、分岐メガロ糖が有する分岐構造は、グルコースがα－１，３－結合により分岐した構造、マルトースがα－１，３－結合により分岐した構造、ニゲロースがα－１，３－結合により分岐した構造、マルトトリオースがα－１，３－結合により分岐した構造、マルトシル－α－１，３－グルコースがα－１，３－結合により分岐した構造、ニゲロシル－α－１，４－グルコースがα－１，３－結合により分岐した構造、ニゲロトリオースがα－１，３－結合により分岐した構造が挙げられる。４糖以上の分岐構造が結合する場合には、その分岐構造は、基質の直鎖状グルカンの非還元末端にα－１，３－結合により結合するグルカンであって、分岐構造を構成するグルコシド結合がα－１，４－結合および／またはα－１，３－結合からなるグルカンであってもよい。 In the production method according to the present invention, when α-glucosidase derived from Acremonium sp is used as a glycosyltransferase, a branched megalosaccharide in which a glucose residue is bonded to the non-reducing end by an α-1,3-glucoside bond can be produced. it can. In this case, the branched structure of the branched megalosaccharide includes a structure in which glucose is branched by α-1,3-bonds, a structure in which maltose is branched by α-1,3-bonds, and nigerose is formed by α-1,3-bonds. A branched structure, a structure in which maltotriose is branched by an α-1,3-bond, a structure in which maltosyl-α-1,3-glucose is branched by an α-1,3-bond, nigerosyl-α-1,4- Examples include a structure in which glucose is branched by an α-1,3-bond, and a structure in which nigerotriose is branched by an α-1,3-bond. When a branched structure of 4 or more sugars is bonded, the branched structure is a glucan that is bonded to the non-reducing end of the linear glucan of the substrate by an α-1,3-linkage, and is a glucoside constituting the branched structure The bond may be a glucan composed of α-1,4-bond and / or α-1,3-bond.
　本発明による製造方法においてアクレモニウム エスピー由来のα－グルコシダーゼを使用した場合には、本発明による分岐メガロ糖を高収率で製造することができ、特に、重合度１５～３５の比較的重合度が高い分岐メガロ糖を高効率で製造することができる。 When α-glucosidase derived from Acremonium sp. Is used in the production method according to the present invention, the branched megalosaccharide according to the present invention can be produced in a high yield, and in particular, a relatively high degree of polymerization with a degree of polymerization of 15 to 35. Therefore, it is possible to produce a highly branched megalosaccharide with high efficiency.
　本発明による製造方法では、また、糖転移酵素としてアスペルギルス ニガー由来のα－グルコシダーゼを使用すると、グルコース残基がα－１，６－グルコシド結合により非還元末端に結合した分岐メガロ糖を製造することができる。この場合、分岐メガロ糖が有する分岐構造は、グルコースがα－１，６－結合により分岐した構造、マルトースがα－１，６－結合により分岐した構造、イソマルトースがα－１，６－結合により分岐した構造、マルトトリオースがα－１，６－結合により分岐した構造、イソパノースがα－１，６－結合により分岐した構造、パノースがα－１，６－結合により分岐した構造、イソマルトトリオースがα－１，６－結合により分岐した構造が挙げられる。４糖以上の分岐構造が結合する場合には、その分岐構造は、基質の直鎖状グルカンの非還元末端にα－１，６－結合により結合するグルカンであって、分岐構造を構成するグルコシド結合がα－１，４－結合および／またはα－１，６－結合からなるグルカンであってもよい。なお、アスペルギルス ニガー由来のα－グルコシダーゼを用いた場合はごく微量ではあるがα－１，２－結合やα－１，３－結合が分岐構造中に含まれることがある。 In the production method according to the present invention, when α-glucosidase derived from Aspergillus niger is used as a glycosyltransferase, a branched megalosaccharide in which a glucose residue is bonded to a non-reducing end by an α-1,6-glucoside bond is produced. Can do. In this case, the branched structure of the branched megalosaccharide includes a structure in which glucose is branched by an α-1,6-bond, a structure in which maltose is branched by an α-1,6-bond, and an isomaltose in an α-1,6-bond. Branched structure, maltotriose branched by α-1,6-bond, isopanose branched by α-1,6-bond, panose branched by α-1,6-bond, An example is a structure in which maltotriose is branched by α-1,6-bonds. When a branched structure of 4 or more sugars is bonded, the branched structure is a glucan that is bonded to the non-reducing end of the linear glucan of the substrate by an α-1,6-bond, and the glucoside constituting the branched structure The bond may be a glucan having an α-1,4-bond and / or an α-1,6-bond. When α-glucosidase derived from Aspergillus niger is used, α-1,2-bonds and α-1,3-bonds may be included in the branched structure, although in very small amounts.
　糖転移酵素として６－α－グルコシルトランスフェラーゼを使用した場合には、直鎖状グルカンの非還元末端にα－１，６－結合によりグルコースが１分子結合した分岐メガロ糖が得られる。 When 6-α-glucosyltransferase is used as a glycosyltransferase, a branched megalosaccharide in which one molecule of glucose is bonded to the non-reducing end of a linear glucan by α-1,6-linkage is obtained.
　糖転移酵素としてデキストリンデキストラナーゼを使用した場合には、直鎖状グルカンの非還元末端にグルコース１～６残基からなる分岐構造がα－１，６－結合により結合した分岐メガロ糖が得られる。この分岐構造のグルカンは主としてα－１，６－結合から構成されるが、わずかにα－１，４－結合が含まれることがある。 When dextrin dextranase is used as a glycosyltransferase, a branched megalosaccharide in which a branched structure consisting of 1 to 6 glucose residues is bonded to the non-reducing end of a linear glucan by an α-1,6-bond is obtained. It is done. This branched glucan is mainly composed of α-1,6-bonds, but may contain a few α-1,4-linkages.
　糖転移酵素として環状マルトシルマルトース生成酵素を使用した場合は、直鎖状グルカンの非還元末端にマルトースまたはマルトシル－α－１，６－マルトースがα－１，６－結合により結合した分岐メガロ糖が得られる。 When a cyclic maltosyl maltose-producing enzyme is used as a glycosyltransferase, a branched megalosaccharide in which maltose or maltosyl-α-1,6-maltose is bonded to the non-reducing end of a linear glucan by an α-1,6-linkage Is obtained.
　本発明では、酵素反応により得られた生成物を、そのまま分岐メガロ糖製品とすることもできる。また、必要に応じて、酵素反応により得られた生成物を遠心分離あるいは濾過等により不溶物を除去し、水溶性画分を濃縮することで、目的とする本発明による分岐メガロ糖の溶液を得ることもできる。あるいは、必要に応じて活性炭により脱色させたもの、適当なイオン交換樹脂によりイオン性成分を除去したものを濃縮してもよい。保存性やその後の用途においては、脱色、イオン除去したものを微生物の繁殖が問題とならない程度の水分活性となるまで濃縮することが好適である。または、用途によっては利用しやすいように、乾燥し、粉末として得ることもできる。乾燥は、通常、凍結乾燥あるいは噴霧乾燥やドラム乾燥などの方法が利用できる。乾燥物は、必要により粉砕することが望ましい。 In the present invention, the product obtained by the enzymatic reaction can be directly used as a branched megalosaccharide product. Further, if necessary, the product obtained by the enzyme reaction is centrifuged or filtered to remove insoluble matters, and the water-soluble fraction is concentrated, whereby the target branched megalosaccharide solution according to the present invention is obtained. It can also be obtained. Or what was decolored with activated carbon as needed, and what removed the ionic component with the appropriate ion exchange resin may be concentrated. In the storage and subsequent use, it is preferable to concentrate the decolorized and ion-removed product until the water activity is such that the propagation of microorganisms does not become a problem. Or it can also be dried and obtained as a powder so that it may be easy to use depending on a use. For drying, a method such as freeze drying, spray drying or drum drying can be used. The dried product is desirably pulverized as necessary.
　従って、本発明によれば、本発明による分岐メガロ糖を固形分当たり１０～６０質量％、好ましくは、１５～５５質量％含有する液糖および粉糖が提供される。なお、本明細書において「粉糖」は粉飴を含む意味で用いられるものとする。 Therefore, according to the present invention, there are provided liquid sugars and powdered sugars containing 10 to 60% by mass, preferably 15 to 55% by mass, of the branched megalosaccharide according to the present invention per solid content. In addition, in this specification, "powder sugar" shall be used with the meaning containing powdered rice cake.
　本発明の酵素反応により得られる生成物は、通常、本発明による分岐メガロ糖と共に少量のシクロデキストリン、デキストリン、または重合度１０以下のオリゴ糖、あるいはこれらの混合物を含有している。この生成物はそのまま後述するような食品などの用途に用いることができるが、必要に応じてこれらの成分を除去し、精製した分岐メガロ糖を得てもよい。あるいは、これらの生成物から重合度５未満のオリゴ糖を除去してもよい。分岐メガロ糖の単離・精製方法およびオリゴ糖の分離・除去方法としては、ゲルろ過クロマトグラフィーなど当業者に周知の糖類の精製方法を使用できる。なお、本発明の酵素反応により得られる生成物には、デンプン原料に由来する分岐構造を非還元末端に加えて直鎖状グルカンの中間部分、すなわち、末端残基以外の糖残基、に有する分岐メガロ糖が僅かに含まれており、そのような分岐メガロ糖も本発明の範囲内であることはいうまでもない。 The product obtained by the enzymatic reaction of the present invention usually contains a small amount of cyclodextrin, dextrin, oligosaccharide having a polymerization degree of 10 or less, or a mixture thereof, together with the branched megalosaccharide according to the present invention. This product can be used as it is for foods and the like as described later, but if necessary, these components may be removed to obtain a purified branched megalosaccharide. Alternatively, oligosaccharides having a polymerization degree of less than 5 may be removed from these products. As a method for isolating / purifying branched megalosaccharides and a method for separating / removing oligosaccharides, saccharide purification methods well known to those skilled in the art, such as gel filtration chromatography, can be used. In addition, the product obtained by the enzyme reaction of the present invention has a branched structure derived from the starch raw material in the middle part of the linear glucan in addition to the non-reducing end, that is, the sugar residue other than the terminal residue. Needless to say, the branched megalosaccharide is slightly contained, and such a branched megalosaccharide is also within the scope of the present invention.
　このようにして得られる本発明による分岐メガロ糖は、その溶液を低温下あるいは固形分７５質量％程度の高濃度溶液として放置しても通常のデキストリンと比較して、老化による白濁が観察されず、顕著な耐老化性を有するという特徴を有する。また、固形分７５質量％程度の溶液状態での粘性が低く、操作性に優れるという特徴も有する。また、糖化反応後のグルコースを始めとする低分子オリゴ糖成分の含有量が少なく、水飴としては極めて低甘味であることも特徴である。 The branched megalosaccharide according to the present invention thus obtained does not show turbidity due to aging compared to ordinary dextrin even when the solution is left at a low temperature or as a high-concentration solution having a solid content of about 75% by mass. It has the characteristic of having remarkable aging resistance. Moreover, it has the characteristics that the viscosity in the solution state of about 75 mass% of solid content is low and it is excellent in operativity. In addition, the content of low-molecular-weight oligosaccharide components such as glucose after saccharification reaction is small, and the varicella is also characterized by extremely low sweetness.
　また、本発明による分岐オリゴ糖を含有する液糖および粉糖であって、重合度４以下のオリゴ糖を除去した液糖および粉糖は、耐老化性を有することはもちろんのこと、低甘味であるとともに、風味改善に用いることができる点で有利である。 Further, liquid sugars and powdered sugars containing branched oligosaccharides according to the present invention from which oligosaccharides having a polymerization degree of 4 or less have been removed have not only aging resistance but also low sweetness. And is advantageous in that it can be used for flavor improvement.
分岐メガロ糖の用途
［食品への用途］
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、食品に添加して使用することができる。従って、本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を含んでなる食品添加剤が提供される。本発明による本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を添加することができる食品は特に限定されないが、例示すれば、茶系飲料、果実飲料、炭酸飲料、野菜飲料、スポーツ飲料、乳性飲料、アルコール飲料、その他の飲料や、アイスクリーム、氷菓、ゼリー、ムース、飴菓子、ガム、フィリング、健康食品、サプリメント、パン類、クッキー類、米飯、ケーキ類、麺類、冷凍食品、凍結飲料が挙げられる。 Use of branched megalo sugar [ use for food]
The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can be added to foods. Therefore, according to the present invention, a food additive comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same is provided. The branched megalosaccharide according to the present invention and the food to which the liquid sugar and powdered sugar containing it can be added are not particularly limited. For example, tea beverages, fruit beverages, carbonated beverages, vegetable beverages, sports Beverages, milky beverages, alcoholic beverages, other beverages, ice cream, ice confectionery, jelly, mousse, candy sweets, gum, fillings, health foods, supplements, breads, cookies, rice, cakes, noodles, frozen foods And frozen beverages.
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、風味改善効果や食感改善効果を有するとともに、低甘味という特徴を有する。従って、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を食品に添加することにより、食品本来の風味を損なわずに食品の風味や食感を改善できる点で有利である。 The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same have a taste improving effect and a texture improving effect, and are characterized by low sweetness. Therefore, by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to the food, it is advantageous in that the flavor and texture of the food can be improved without impairing the original flavor of the food.
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、また、添加物が溶解または分散した水を均一に凍結させる効果を有する。従って、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を、添加物が溶解または分散した水に添加することにより、添加物が均一に分散した氷を製造できる点で有利である。 The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same also have an effect of uniformly freezing water in which the additive is dissolved or dispersed. Therefore, it is advantageous in that ice having a uniformly dispersed additive can be produced by adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same to water in which the additive is dissolved or dispersed. .
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、また、食品の照りやつやを向上させることができる。従って、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を食品に添加することにより、照りやつやが向上した商品を提供できる点で有利である。 The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it can also improve the shine and gloss of food. Therefore, by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it to the food, it is advantageous in that a product with improved shine and gloss can be provided.
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、また、乳タンパク質の凝集や沈殿を抑制することができる。従って、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を乳タンパク質含有飲料に添加することにより、乳タンパク質の凝集や沈殿が抑制され、乳タンパク質が安定化された乳タンパク質含有飲料を提供できる点で有利である。 The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can also suppress milk protein aggregation and precipitation. Therefore, by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to the milk protein-containing beverage, milk protein aggregation and precipitation are suppressed, and the milk protein-containing beverage is stabilized. Is advantageous in that it can be provided.
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖の食品への添加方法は当業者であればその食品の製造方法に従って適宜選択することができ、食品の製造原料に予め配合して製造しても、食品の製造工程中あるいは製造後に配合して製造してもよい。 A person skilled in the art can appropriately select a branched megalosaccharide and liquid sugar and powdered sugar containing the same according to the present invention according to the method for producing the food. Even if it manufactures, it may mix | blend and manufacture in the manufacturing process of a foodstuff, or after manufacture.
［医薬品への用途］
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、また、医薬品に添加して使用することができる。従って、本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を含んでなる製剤用添加剤が提供される。本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を添加することができる医薬品は特に限定されないが、例えば、経口投与用製剤が挙げられ、好ましくは、錠剤、散剤、顆粒剤、細粒剤、液剤、シロップ剤（ドライシロップ剤を含む）、カプセル剤、トローチ剤などである。 [Use for pharmaceutical products]
The branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same can also be used by adding to pharmaceuticals. Therefore, according to the present invention, there are provided a pharmaceutical additive comprising the branched megalosaccharide according to the present invention, and liquid sugar and powdered sugar containing the same. There are no particular limitations on the medicinal product to which the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing it can be added, and examples thereof include preparations for oral administration, preferably tablets, powders, granules, fine powders, and the like. Granules, liquids, syrups (including dry syrups), capsules, lozenges and the like.
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、風味改善効果を有する。従って、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を医薬品に添加することにより、有効成分や他の添加成分の味を矯正あるいはマスキングできるとともに、賦形剤としても利用できる点で有利である。 The branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same have a flavor improving effect. Therefore, by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it to a pharmaceutical product, the taste of the active ingredient and other additive ingredients can be corrected or masked, and can also be used as an excipient. Is advantageous.
　医薬製剤の製造方法や医薬品の原料となる製剤用添加剤は当業者に周知であり、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を、有効成分や他の製剤用添加剤と混合等することにより、常法に従って製造することができる。 Pharmaceutical preparation methods and pharmaceutical additives as raw materials for pharmaceuticals are well known to those skilled in the art, and branched megalosaccharides according to the present invention and liquid sugars and powdered sugars containing them are used as active ingredients and other pharmaceutical additives. Can be produced in accordance with a conventional method.
［風味の改善］
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、後述する配合例に示すように、食品等の風味を改善することができる。従って、本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を含んでなる風味改善剤および製剤用マスキング剤が提供される。 [Improvement of flavor]
The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can improve the flavor of foods and the like as shown in the formulation examples described later. Therefore, according to the present invention, there are provided a flavor-improving agent and a masking agent for preparations comprising the branched megalosaccharide according to the present invention, and liquid sugar and powdered sugar containing the same.
　ここで、風味改善としては、食品や食品添加物に特有の不快な味や臭いの改善が挙げられ、例えば、苦味および／または渋味の低減（例えば、ポリフェノール類の苦味や渋味の低減）、酸味の低減（例えば、酢や酸味剤に由来する酸味の緩和）、エグ味および／または嫌味の低減（例えば、グルコン酸など保存剤の不快な味の低減）、アルコール感の低減（例えば、ウォッカ、ウイスキーなどのアルコール感の低減）、酸臭その他不快臭の低減（例えば、グルコノデルタラクトン臭（グルコン酸臭）やデキストリン臭の低減）が挙げられる。風味改善としては、また、コク味の増強（例えば、果汁感や乳味感の増強）が挙げられる。本発明による風味改善剤により風味が改善される物としては、前記の食品が挙げられる。 Here, examples of flavor improvement include improvement of unpleasant taste and odor peculiar to foods and food additives, for example, reduction of bitterness and / or astringency (for example, reduction of bitterness and astringency of polyphenols). , Reduction of acidity (e.g., mitigation of acidity derived from vinegar and acidulants), reduction of egg taste and / or taste (e.g., reduction of unpleasant taste of preservatives such as gluconic acid), reduction of alcoholic feeling (e.g., Reduction of alcohol feeling such as vodka and whiskey), reduction of acid odor and other unpleasant odor (for example, reduction of glucono delta lactone odor (gluconic acid odor) and dextrin odor). Examples of the flavor improvement include enhancement of richness (for example, enhancement of fruit juice feeling and milky feeling). Examples of the product whose flavor is improved by the flavor improving agent according to the present invention include the foods described above.
　また、医薬製剤において矯正あるいはマスキングされる味としては、有効成分や他の製剤用添加剤に特有の不快な味（例えば、苦味、渋味、酸味、エグ味）が挙げられる。 In addition, the taste corrected or masked in the pharmaceutical preparation includes an unpleasant taste (for example, bitter taste, astringency, sour taste, and savory taste) peculiar to active ingredients and other additives for pharmaceutical preparations.
　本発明による分岐メガロ糖は、苦味および／または渋味を効果的に低減することから、本発明による風味改善剤は、好ましくは、苦味および／または渋味の低減剤として用いることができる。 Since the branched megalosaccharide according to the present invention effectively reduces bitterness and / or astringency, the flavor improving agent according to the present invention can be preferably used as a bitterness and / or astringency reducing agent.
　特に、本発明による分岐メガロ糖は、ポリフェノール類に起因する苦味および／または渋味を効果的に低減することから、本発明による風味改善剤は、好ましくは、ポリフェノール類の苦味および／または渋味の低減剤として用いることができる。 In particular, since the branched megalosaccharide according to the present invention effectively reduces bitterness and / or astringency caused by polyphenols, the flavor improving agent according to the present invention is preferably a bitterness and / or astringency of polyphenols. It can be used as a reducing agent.
　ポリフェノール類とは分子内に複数のフェノール性ヒドロキシル基を持つ成分を意味し、例えば、カテキン、アントシアニン、クロロゲン酸などが挙げられる。ポリフェノール類には生体に好ましい様々な生理機能が知られているが、ほとんどのポリフェノール類は強い苦味や渋味を呈するため、高濃度の摂取が困難である。本発明による分岐メガロ糖はポリフェノール類に起因する苦味および／または渋味を効果的に低減することから、本発明による風味改善剤はポリフェノール類の高濃度摂取を可能にする点で有利である。 Polyphenols mean components having a plurality of phenolic hydroxyl groups in the molecule, and examples thereof include catechin, anthocyanin, chlorogenic acid and the like. Polyphenols are known to have various physiological functions favorable to living organisms, but most polyphenols exhibit strong bitterness and astringency, making it difficult to ingest high concentrations. Since the branched megalosaccharide according to the present invention effectively reduces the bitterness and / or astringency caused by polyphenols, the flavor improving agent according to the present invention is advantageous in that it allows high-concentration intake of polyphenols.
　本発明による分岐メガロ糖はまた、低甘味であることから、本発明による風味改善剤を食品に添加した場合でも、食品本来の風味を損なわずに風味を改善できる点で有利である。 The branched megalosaccharide according to the present invention is also low in sweetness, which is advantageous in that the flavor can be improved without impairing the original flavor of the food even when the flavor improver according to the present invention is added to the food.
　本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を食品に添加することを含んでなる、風味の改善方法および風味が改善された食品の製造方法が提供される。 According to the present invention, there is provided a method for improving flavor and a method for producing food with improved flavor, comprising adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same to food. The
　本発明によれば、また、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を苦味および／または渋味を有する食品に添加することを含んでなる、苦味および／または渋味の低減化方法並びに苦味および／または渋味が低減された食品の製造方法が提供される。前述のように、本発明による分岐メガロ糖は、ポリフェノール類に由来する苦味および／または渋味を効果的に低減することから、苦味および／または渋味を有する食品としては、ポリフェノール類含有食品が挙げられる。 According to the present invention, the bitter and / or astringent taste comprising the addition of the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to foods having a bitter and / or astringent taste. Provided are a reduction method and a method for producing a food with reduced bitterness and / or astringency. As described above, the branched megalosaccharide according to the present invention effectively reduces the bitterness and / or astringency derived from polyphenols. Therefore, as a food having a bitterness and / or astringency, a polyphenols-containing food is used. Can be mentioned.
　ポリフェノール類含有食品としては、ポリフェノール類を含有するものであれば特に限定されず、元々ポリフェノール類を含有している食品はもちろんのこと、ポリフェノール類が添加された食品も該当する。また、食品の形態も特に限定されず、固形はもちろんのこと、半固形、液状のものも含まれる。ポリフェノール類含有食品の具体例としては、茶系飲料（緑茶飲料、紅茶飲料、ウーロン茶飲料など）、果実飲料、炭酸飲料、野菜飲料、スポーツ飲料、乳性飲料、アルコール飲料などの飲料や、アイスクリーム、ゼリー、ムース、飴菓子、ガム、フィリング、健康食品、サプリメントなどが挙げられる。なお、本発明において「食品」とは飲料も含む意味で用いられるものとする。 The polyphenol-containing food is not particularly limited as long as it contains polyphenols, and foods to which polyphenols are added as well as foods originally containing polyphenols are also applicable. Also, the form of the food is not particularly limited, and includes not only solids but also semisolids and liquids. Specific examples of polyphenol-containing foods include beverages such as tea beverages (green tea beverages, tea beverages, oolong tea beverages, etc.), fruit beverages, carbonated beverages, vegetable beverages, sports beverages, milk beverages, alcoholic beverages, and ice creams. , Jelly, mousse, candy, gum, filling, health food, supplements and the like. In the present invention, the term “food product” is used to include beverages.
　本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を、医薬品に添加することを含んでなる、不快な味のマスキング方法または不快な味がマスキングされた医薬品の製造方法が提供される。 According to the present invention, there is provided an unpleasant taste masking method or an unpleasant taste masked pharmaceutical comprising adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to the pharmaceutical. A manufacturing method is provided.
［食感の改善］
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、また、食品等の食感を改善することができる。従って、本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を含んでなる食感改善剤が提供される。 [Improve texture]
The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can also improve the texture of foods and the like. Therefore, according to the present invention, there is provided a texture improving agent comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same.
　ここで、食感改善としては、パサパサ感の抑制、しっとり感の付与、ふんわりした食感の維持が挙げられる。本発明による風味改善剤により食感が改善される物としては、前記の食品のうち食感の改善が求められる食品が挙げられ、そのような食品の例としては、パン類、クッキー類、米飯、ケーキ類、麺類、冷凍食品が挙げられる。 Here, examples of the texture improvement include suppression of a feeling of dryness, imparting a moist feeling, and maintaining a soft texture. Examples of the food whose texture is improved by the flavor improving agent according to the present invention include foods whose food texture is required to be improved, and examples of such foods include breads, cookies, and cooked rice. , Cakes, noodles, frozen foods.
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は食感改善を目的として食品に添加することができるが、前記の風味改善をも目的として添加してもよいことは言うまでもない。例えば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を米飯に添加した場合には、パサつきの防止という食感改善効果が得られるが、米飯に添加される日持ち向上剤（例えば、グルコノデルタラクトンやグルコン酸）など他の添加剤の嫌味や不快臭を低減することが可能となる。 The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it can be added to foods for the purpose of improving the texture, but it goes without saying that they may also be added for the purpose of improving the flavor. For example, when the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same are added to the cooked rice, the texture improving effect of preventing the puffiness is obtained, but the shelf life improver added to the cooked rice (for example, , Glucono delta lactone and gluconic acid) and the like can be reduced.
　本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を食品に添加することを含んでなる、食感の改善方法および食感が改善された食品の製造方法が提供される。 According to the present invention, there is provided a method for improving texture and a method for producing food with improved texture, comprising adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same to food. Provided.
［氷の均一的凍結］
　添加物を含有する水を凍結させると、氷結晶は添加物を排除しながら成長するため添加物が偏在する氷ができる。このような添加物が偏在した氷は、色、味付け、食感などに偏りやむらが生じており、商品価値を低下させることになる。 [Uniform freezing of ice]
When water containing the additive is frozen, ice crystals grow while excluding the additive, so that ice with an ubiquitous additive is formed. Such unevenly distributed ice has unevenness and unevenness in color, seasoning, texture, and the like, thereby reducing the commercial value.
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、添加物が溶解または分散した水を均一に凍結させることができる。従って、本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を含んでなる、氷の均一的凍結剤が提供される。本発明による氷の均一的凍結剤を用いて添加物を含有する水を凍結させることにより、偏りやむらのない均一な色彩、食感、味覚等を有する氷菓を製造することができる。 The branched megalosaccharide and liquid sugar and powdered sugar containing the same according to the present invention can uniformly freeze water in which the additive is dissolved or dispersed. Therefore, according to the present invention, there is provided a uniform ice freezing agent comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same. By freezing the water containing the additive using the ice uniform freezing agent according to the present invention, frozen dessert having uniform color, texture, taste, etc. without unevenness and unevenness can be produced.
　ここで、氷の「均一的凍結」とは、添加物を含有する水を添加物が均一に溶解または分散した状態で凍結させることを意味する。 Here, “uniform freezing” of ice means freezing water containing the additive in a state where the additive is uniformly dissolved or dispersed.
　また、水に含まれる「添加物」としては、糖類、塩類、果汁、色素、植物エキス、茶、コーヒー、香料、香辛料、タンパク質、アミノ酸、ゲル化剤、酸味料、食物繊維などが挙げられる。なお、「添加物が溶解または分散した水」には、果汁そのものや植物エキスそのものなど、水以外の成分が元々混合している溶液をも含む意味で用いられることは言うまでもない。 In addition, examples of “additives” contained in water include sugars, salts, fruit juices, pigments, plant extracts, tea, coffee, flavorings, spices, proteins, amino acids, gelling agents, acidulants, and dietary fibers. Needless to say, “water in which an additive is dissolved or dispersed” includes a solution in which components other than water, such as fruit juice itself and plant extract itself, are originally mixed.
　本発明によれば、添加物が均一に溶解または分散した水に、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を添加して凍結させることを含んでなる、氷の均一的凍結方法または均一的に凍結した氷の製造方法が提供される。 According to the present invention, the homogeneous ice is obtained, which comprises freezing the water in which the additive is uniformly dissolved or dispersed by adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same. A freezing method or a method for producing uniformly frozen ice is provided.
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を添加することにより、添加物が均一に溶解または分散した状態で水を凍結させることができる。従って、例えば、得られた氷を冷菓とすれば、偏りやむらのない均一な色彩、食感、味覚等を有する氷菓を提供することができる。また、本発明による氷の均一的凍結剤を、果汁飲料やスポーツ飲料などの飲料に添加して凍結させれば、それらを一部解凍した場合でも均一な溶液となる飲料を提供することができる。 By adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same, water can be frozen in a state where the additive is uniformly dissolved or dispersed. Therefore, for example, when the obtained ice is used as a frozen dessert, an iced dessert having a uniform color, texture, taste, etc. without unevenness and unevenness can be provided. In addition, if the ice uniform freezing agent according to the present invention is added to a beverage such as a fruit juice beverage or a sports beverage and frozen, a beverage that becomes a uniform solution even when partly thawed can be provided. .
　本発明による氷の製造方法により製造された氷は、そのままで、あるいは加工して、例えば、シャーベット、かき氷、かちわり氷、アイスキャンディー、シェイク、ソルベットなどの氷菓；果汁飲料、スポーツ飲料、アミノ酸飲料などの飲料；ゼリーやヨーグルトなどの冷菓子類として提供することができる。 The ice produced by the method for producing ice according to the present invention is used as it is or after processing, for example, ice confectionery such as sherbet, shaved ice, flake ice, ice candy, shake, sorbet; fruit juice drink, sports drink, amino acid drink Beverages such as jelly and yogurt.
［照り・つやの向上］
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、また、食品の照りやつやを向上させることができる。従って、本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を含んでなる、食品の照りおよび／またはつや向上剤が提供される。ここで、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖により食品の照りおよび／またはつやの向上が期待される食品としては食肉が挙げられ、好ましくは、鶏肉である。 [Improve shine and gloss]
The branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it can also improve the shine and gloss of food. Therefore, according to the present invention, there is provided a food shine and / or luster improving agent comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same. Here, meat is mentioned as a food in which the shimmering and / or improvement in gloss of the food is expected by the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same, and preferably chicken.
　本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を食品に添加することを含んでなる、食品の照りおよび／またはつやの向上方法または照りおよび／またはつやが向上した食品の製造方法が提供される。これらの方法では、例えば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を食品に予め塗布し、その食品を焼成してもよい。食品を焼成した後、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を再度塗布して焼成してもよい。例えば、照り焼きを提供する場合には、照り焼きのタレに本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を添加し、そのタレに食品を浸漬した後、焼成してもよい。あるいは、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を焼成前に食品の表面に直接吹き付けて使用してもよい。 According to the present invention, the method for improving the shine and / or shine of the food comprising adding the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same to the food is improved. There is provided a method for producing a prepared food. In these methods, for example, the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same may be applied to the food in advance, and the food may be baked. After the food is baked, the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same may be applied again and baked. For example, when providing teriyaki, the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the teriyaki sauce may be added, and the food may be immersed in the sauce and then baked. . Alternatively, the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it may be directly sprayed on the surface of the food before baking.
［乳タンパク質含有飲料への添加］
　本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、乳タンパク質含有飲料に添加して使用することができる。本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を乳タンパク質含有飲料に添加しても乳タンパク質が凝集・沈殿せず、乳タンパク質を飲料中において安定化させることができる。すなわち、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を乳タンパク質含有飲料に使用すると乳タンパク質の凝集・沈殿を回避することができる。また、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、単糖、二糖、オリゴ糖等と比較して、溶液中で低浸透圧である。従って、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖は、好ましくは、乳タンパク質を含有する経腸栄養剤や経口栄養剤に添加して使用することができる。 [Addition to milk protein-containing beverages]
The branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same can be used by adding to a milk protein-containing beverage. Even when the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it are added to a milk protein-containing beverage, the milk protein does not aggregate and precipitate, and the milk protein can be stabilized in the beverage. That is, when the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it are used in a milk protein-containing beverage, aggregation and precipitation of milk protein can be avoided. Moreover, the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing it have a lower osmotic pressure in the solution than monosaccharide, disaccharide, oligosaccharide and the like. Therefore, the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same can be preferably used by being added to an enteral nutrition or oral nutrition containing milk protein.
　本発明によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を乳タンパク質含有飲料に添加することを含んでなる、乳タンパク質の凝集および／または沈殿が抑制された乳タンパク質含有飲料の製造方法が提供される。これらの方法における本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖の添加時期は特に限定されず、乳タンパク質と一緒に添加しても、乳タンパク質とは別に添加してもよい。 According to the present invention, milk protein in which aggregation and / or precipitation of milk protein is suppressed, comprising adding the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same to a milk protein-containing beverage. A method for producing a beverage containing beverage is provided. The addition timing of the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same in these methods is not particularly limited, and may be added together with milk protein or separately from milk protein.
［食品、医薬品等］
　本発明による食品添加剤や本発明による風味改善剤、食感改善剤、氷の均一的凍結剤、照りおよび／またはつや向上剤、および乳タンパク質の凝集および／または沈殿抑制剤は、食品に添加されて使用できる。従って、本発明の別の面によれば、本発明による食品添加剤や本発明による風味改善剤、食感改善剤、氷の均一的凍結剤、照りおよび／またはつや向上剤、または乳タンパク質の凝集および／または沈殿抑制剤を含んでなる食品が提供される。 [Food, pharmaceuticals, etc.]
The food additive according to the present invention, the flavor improving agent, the texture improving agent, the ice uniform freezing agent, the shine and / or gloss improving agent, and the milk protein aggregation and / or precipitation inhibitor are added to the food. Can be used. Therefore, according to another aspect of the present invention, the food additive according to the present invention, the flavor improving agent, the texture improving agent, the ice uniform freezing agent, the shine and / or the gloss improving agent, or the milk protein according to the present invention. A food product comprising an agglomeration and / or precipitation inhibitor is provided.
　本発明の更に別の面によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を含んでなる食品が提供される。 According to still another aspect of the present invention, there is provided a food comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same.
　この場合、提供される食品は、好ましくは、ポリフェノール類含有食品であってもよく、具体例や好ましい例は前述の通りである。 In this case, the food provided may be a polyphenol-containing food, and specific examples and preferred examples are as described above.
　ポリフェノール類含有食品に本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を使用する場合には、例えば、甘味を必要とする飲食物を対象とする場合、本発明による分岐メガロ糖に砂糖、異性化糖、高甘味度甘味料といった甘味の強い組成物を混合して使用することができる。一方、緑茶飲料や惣菜といった甘味の好まれない飲食物に使用する場合は分岐メガロ糖単独もしくは分岐メガロ糖と多糖やタンパク質、油脂等を混合して使用することができる。 When the branched megalosaccharide according to the present invention and the liquid sugar and powdered sugar containing the same are used for polyphenols-containing foods, for example, when targeting foods and drinks that require sweetness, A composition having a strong sweetness such as sugar, isomerized sugar, and a high-intensity sweetener can be mixed and used. On the other hand, when used for foods and beverages that do not like sweet taste such as green tea beverages and side dishes, it is possible to use branched megalosaccharides alone or a mixture of branched megalosaccharides and polysaccharides, proteins, fats and the like.
　本発明による食品の好ましい態様によれば、ポリフェノール類０．１３質量％に対して、本発明による分岐メガロ糖を１．６～５０質量％含有するポリフェノール類含有食品が提供される。 According to a preferred aspect of the food according to the present invention, there is provided a polyphenol-containing food containing 1.6 to 50% by mass of the branched megalosaccharide according to the present invention with respect to 0.13% by mass of the polyphenols.
　また、提供される食品は乳タンパク質含有飲料であってもよく、より好ましくは、乳タンパク質を含有する経腸栄養剤や経口栄養剤である。 Further, the food to be provided may be a milk protein-containing beverage, more preferably an enteral nutrient or an oral nutrient containing milk protein.
　本発明による製剤用添加剤や製剤用マスキング剤は医薬品に配合されて使用できる。従って、本発明の別の面によれば、本発明による製剤用添加剤や本発明による製剤用マスキング剤を含んでなる医薬品が提供される。 The formulation additive and the formulation masking agent according to the present invention can be used in combination with a pharmaceutical product. Therefore, according to another aspect of the present invention, there is provided a pharmaceutical comprising the pharmaceutical additive according to the present invention and the pharmaceutical masking agent according to the present invention.
　本発明の更に別の面によれば、本発明による分岐メガロ糖並びにそれを含有する液糖および粉糖を含んでなる医薬品が提供される。 According to still another aspect of the present invention, there is provided a medicinal product comprising the branched megalosaccharide according to the present invention and liquid sugar and powdered sugar containing the same.
　以下の例に基づいて本発明を具体的に説明するが、本発明はこれらの例に限定されるものではない。なお、本明細書において「固形分」当たりの割合や「固形分」の含有割合に言及した場合には、固形成分の質量に基づいて定められた割合を意味するものとする。 The present invention will be specifically described based on the following examples, but the present invention is not limited to these examples. In the present specification, when the ratio per “solid content” or the content ratio of “solid content” is mentioned, it means the ratio determined based on the mass of the solid component.
試験例１：糖化酵素の活性測定
１－１：シクロデキストリン生成酵素の活性測定
　糖化反応に使用したシクロデキストリン生成酵素（ＣＧＴａｓｅ）を以下に示す。
・パエニバチルス　エスピー由来のＣＧＴａｓｅ：ナガセケムテックス社製アルカリＣＤアミラーゼ
・バチルス　コアギュランス由来のＣＧＴａｓｅ：ナガセケムテックス社製ネオＣＤアミラーゼ
・バチルス　ステアロサーモフィルス由来のＣＧＴａｓｅ：林原生物化学研究所製Ｔｈｅｒｍｏｐｈｉｌｉｃ　ＣＧＴａｓｅ
・バチルス　マゼランス由来のＣＧＴａｓｅ：アマノエンザイム社製のコンチザイム
　酵素反応は、５０ｍＭリン酸カリウム緩衝液（ｐＨ６．０）に溶解した１％可溶性デンプン（ナカライテスク社）０．９ｍｌに適宜水で希釈した酵素溶液０．１ｍｌを添加し、４０℃に１０分間保持した。これに４０ｍＭ水酸化ナトリウム水溶液を２．５ｍｌ添加して反応を停止した。生成したβ－シクロデキストリンをフェノールフタレイン法により測定した。すなわち、０．１ｍｇ／ｍｌフェノールフタレインおよび２．５ｍＭ炭酸ナトリウムからなる溶液０．３ｍｌを上記溶液に添加し、攪拌後５５０ｎｍの吸光度を測定した。０～０．１ｍｇ／ｍｌの範囲で作成したβ－シクロデキストリンの標準曲線に基づき生成したβ－シクロデキストリン量を求めた。 Test Example 1: Measurement of saccharifying enzyme activity
1-1: Activity measurement of cyclodextrin-forming enzyme Cyclodextrin-forming enzyme (CGTase) used in the saccharification reaction is shown below.
-CGTase derived from Paenibacillus sp .: Alkaline CD amylase manufactured by Nagase ChemteX Corp.-CGTase derived from Bacillus coagulans: Neo CD amylase manufactured by Nagase ChemteX Corp.
CGTase derived from Bacillus mazelans: Contizyme manufactured by Amano Enzyme The enzyme reaction was performed by appropriately diluting water with 0.9 ml of 1% soluble starch (Nacalai Tesque) dissolved in 50 mM potassium phosphate buffer (pH 6.0). 0.1 ml of solution was added and held at 40 ° C. for 10 minutes. The reaction was stopped by adding 2.5 ml of 40 mM sodium hydroxide aqueous solution thereto. The produced β-cyclodextrin was measured by the phenolphthalein method. That is, 0.3 ml of a solution composed of 0.1 mg / ml phenolphthalein and 2.5 mM sodium carbonate was added to the above solution, and the absorbance at 550 nm was measured after stirring. The amount of β-cyclodextrin produced was determined based on a standard curve of β-cyclodextrin prepared in the range of 0 to 0.1 mg / ml.
１－２：α－グルコシダーゼの活性測定
　糖化反応に使用したα－グルコシダーゼを以下に示す。
・アスペルギルス　ニガー由来のα－グルコシダーゼ：アマノエンザイム社製トランスグルコシダーゼアマノ
・アクレモニウム　エスピー由来のα－グルコシダーゼ：キリンフードテック社製テイスターゼ
　酵素反応は、５０ｍＭ酢酸ナトリウム緩衝液（ｐＨ４．２）に溶解した０．２５％マルトース８０μｌに０．０５％トリトンＸ－１００を含む１０ｍＭ酢酸ナトリウム緩衝液　（ｐＨ４．２）で適宜希釈した酵素溶液２０μｌを添加し、３７℃に１０分間保持した。
反応１０分で反応液５０μｌを抜き出し、２Ｍトリス塩酸緩衝液（ｐＨ７．０）１００μｌと混合して反応を停止した。これにグルコースＣＩＩ－テストワコー（和光純薬社）を４０μｌ添加した後、室温に１時間保持して発色させ、４９０ｎｍの吸光度を測定した。
生成したグルコース量を０～０．０１％の範囲で作成したグルコースの標準曲線に基づき算出した。 1-2: Measurement of α-glucosidase activity The α-glucosidase used in the saccharification reaction is shown below.
Α-glucosidase derived from Aspergillus niger: transglucosidase manufactured by Amano Enzyme, Amano Acremonium sp-derived α-glucosidase: manufactured by Kirin Foodtech Co., Ltd. The enzyme reaction was dissolved in 50 mM sodium acetate buffer (pH 4.2). 20 μl of an enzyme solution appropriately diluted with 10 mM sodium acetate buffer (pH 4.2) containing 0.05% Triton X-100 was added to 80 μl of 25% maltose, and kept at 37 ° C. for 10 minutes.
In 10 minutes of reaction, 50 μl of the reaction solution was extracted and mixed with 100 μl of 2M Tris-HCl buffer (pH 7.0) to stop the reaction. To this was added 40 μl of glucose CII-Test Wako (Wako Pure Chemical Industries, Ltd.), and the color was developed by keeping at room temperature for 1 hour, and the absorbance at 490 nm was measured.
The amount of produced glucose was calculated based on a standard curve of glucose prepared in the range of 0 to 0.01%.
１－３：イソアミラーゼの活性測定
　糖化反応に使用したイソアミラーゼを以下に示す。
・マイロイデス　オドラータス由来のイソアミラーゼ：合同酒精社製ＧＯＤＯ－ＦＩＡ
・シュードモナス　アミロデラモサ由来のイソアミラーゼ：林原生物科学研究所製のイソアミラーゼ
　反応は、２０ｍＭ塩化カルシウムを含む５０ｍＭ酢酸ナトリウム緩衝液（ｐＨ６．０）１００μｌに５ｍｇ／ｍｌワキシーコーンスターチ（日本食品化工社）３５０μｌを添加し、４５℃に５分間保持したものに同緩衝液にて適宜希釈した酵素溶液１００μｌ添加して４５℃に１５分間保持した。これに反応失活用ヨウ素液（６．３５ｍｇ／ｍｌヨウ素および８３ｍｇ／ｍｌヨウ化カリウムからなる溶液２ｍｌと０．１Ｎ塩酸８ｍｌを混合したもの）５００μｌを添加して反応を停止した。この反応停止液を室温に１５分間保持し、これに純水１０ｍｌ添加したものの６１０ｎｍの吸光度を測定した。 1-3: Measurement of isoamylase activity The isoamylase used in the saccharification reaction is shown below.
-Isoamylase derived from Myroides odoratus: GODO-FIA manufactured by Godo Shusei Co., Ltd.
Pseudomonas amyloderamosa-derived isoamylase: Isoamylase produced by Hayashibara Bioscience Laboratories was prepared by adding 350 mg of 5 mg / ml waxy corn starch (Nippon Food Chemical Co., Ltd.) to 100 μl of 50 mM sodium acetate buffer solution (pH 6.0) containing 20 mM calcium chloride. 100 μl of an enzyme solution appropriately added with the same buffer was added to what was added and kept at 45 ° C. for 5 minutes, and kept at 45 ° C. for 15 minutes. The reaction was stopped by adding 500 μl of a reaction lost iodine solution (a mixture of 2 ml of a solution consisting of 6.35 mg / ml iodine and 83 mg / ml potassium iodide and 8 ml of 0.1N hydrochloric acid). The reaction stop solution was kept at room temperature for 15 minutes, and 10 ml of pure water was added thereto, and the absorbance at 610 nm was measured.
試験例２：分岐メガロ糖の調製
　デンプン液化液にシクロデキストリン生成酵素、α－グルコシダーゼおよび枝切り酵素を作用させることにより得られる反応生成物の構造と物性を調べた。 Test Example 2: Preparation of branched megalosaccharide The structure and physical properties of a reaction product obtained by allowing cyclodextrin-forming enzyme, α-glucosidase and debranching enzyme to act on starch liquefaction liquid were examined.
２－１：シクロデキストリン生成酵素とα－グルコシダーゼによる分岐メガロ糖の生成
　３０質量％ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、アマノエンザイム社製プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アクレモニウム　エスピーのα－グルコシダーゼを対固形分１ｇ当たり０～０．６５単位添加して７２時間糖化した。 2-1 Production of branched megalosaccharide by cyclodextrin-producing enzyme and α-glucosidase 30% by mass DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C and pH 6.0, and this was paired with Paenibacillus sp. Cyclodextrin-producing enzyme. 1 unit per gram of solids, 100 units per gram of Myloides odoratus isoamylase, 1% of Amanoenzyme pullulanase "Amano" 3 per 0.01% of solids, and α-glucosidase of Acremonium sp. Saccharification was carried out for 72 hours by adding 0 to 0.65 unit per 1 g of the minute.
　上記反応液を用いて分岐メガロ糖の生成を以下の方法で確認した。すなわち、５質量％に調整した糖液１ｍＬに１Ｍ酢酸ナトリウム緩衝液（ｐＨ５．５）に溶解した１０ｍｇ／ｍＬ　β－アミラーゼ＃１５００（ナガセケムテックス社）５０μＬを添加し、５５℃にて１時間程度作用させ、煮沸失活させた。これをアンバーライトＭＢ４（オルガノ社）にて脱塩した後、０．４５μｍフィルターにてろ過したものを高速液体クロマトグラフィー（ＨＰＬＣ）に供した。ＨＰＬＣ条件としては、カラムにＭＣＩ　ＧＥＬ　ＣＫ０２ＡＳ
　（φ２０×２５０ｍｍ、三菱化学社）を用い、移動相を超純水とし、カラム温度８５℃、流速１．０ｍｌ／分とした。分析では、２０μＬ程度をクロマトグラフィーに供した。
いずれの条件でも、得られるクロマトグラムのピーク面積より各重合度成分の含有量を求めることができ、重合度１１以上の糖質の含有量を分岐メガロ糖含有量として算出した。 The production of branched megalosaccharide was confirmed by the following method using the reaction solution. That is, 50 μL of 10 mg / mL β-amylase # 1500 (Nagase ChemteX) dissolved in 1 M sodium acetate buffer (pH 5.5) was added to 1 mL of sugar solution adjusted to 5% by mass, and the mixture was added at 55 ° C. for 1 hour. It was allowed to act to a certain extent and was inactivated by boiling. This was desalted with Amberlite MB4 (organo) and then filtered through a 0.45 μm filter for high performance liquid chromatography (HPLC). As HPLC conditions, MCI GEL CK02AS was used in the column.
(Φ20 × 250 mm, Mitsubishi Chemical Co., Ltd.), the mobile phase was ultrapure water, the column temperature was 85 ° C., and the flow rate was 1.0 ml / min. In the analysis, about 20 μL was subjected to chromatography.
Under any condition, the content of each polymerization degree component can be determined from the peak area of the obtained chromatogram, and the content of carbohydrates having a polymerization degree of 11 or more was calculated as the branched megalosaccharide content.
　得られたα－グルコシダーゼ各添加量における反応液にβ－アミラーゼ処理を行ったところ、表１に示したように、α－グルコシダーゼの添加量が多いほど、β－アミラーゼ処理後に残存する重合度１１～３５の分岐メガロ糖画分の含有量が高いことが明らかになった。α－グルコシダーゼ未添加区では、β－アミラーゼ処理後にβ－限界デキストリンと考えられるデキストリン画分のみが残存し、重合度１１～３５のメガロ糖成分はほぼ完全にβ－アミラーゼにより分解した。このことから、α－グルコシダーゼを共存させてシクロデキストリン生成酵素をデンプン液化液に作用させることにより、非還元性末端に分岐構造を有する分岐メガロ糖が得られることが明らかになった。
Figure JPOXMLDOC01-appb-T000001
 （上記表中、表中の成分以外は重合度１０以下の低分子オリゴ糖であり、デキストリンは重合度３５を超える高分子成分を指す。） When the obtained α-glucosidase was subjected to β-amylase treatment for each addition amount of α-glucosidase, as shown in Table 1, as the addition amount of α-glucosidase was increased, the degree of polymerization remaining after β-amylase treatment was 11 It was revealed that the content of ˜35 branched megalo sugar fractions was high. In the group without α-glucosidase, only the dextrin fraction considered to be β-limit dextrin remained after β-amylase treatment, and the megalosaccharide component having a polymerization degree of 11 to 35 was almost completely decomposed by β-amylase. From this, it was revealed that a branched megalosaccharide having a branched structure at the non-reducing end can be obtained by allowing a cyclodextrin-forming enzyme to act on a starch liquefaction solution in the presence of α-glucosidase.
Figure JPOXMLDOC01-appb-T000001
 (In the above table, components other than the components in the table are low-molecular oligosaccharides having a degree of polymerization of 10 or less, and dextrin refers to a polymer component having a degree of polymerization of more than 35.)
　また、図１に示すように、反応７２時間後のα－グルコシダーゼ未添加区では糖化反応中の基質のデンプンの老化が著しく、反応中に白濁する現象が認められたが、α－グルコシダーゼ添加区では反応液の透明性が保たれていた。このことから、分岐メガロ糖は非還元性末端の分岐構造により耐老化性に優れることが確認された。 In addition, as shown in FIG. 1, in the group without addition of α-glucosidase after 72 hours of reaction, the aging of the starch of the substrate during the saccharification reaction was remarkable, and a phenomenon of white turbidity was observed during the reaction. Then, the transparency of the reaction solution was maintained. From this, it was confirmed that the branched megalosaccharide is excellent in aging resistance due to the branched structure of the non-reducing terminal.
２－２：シクロデキストリン生成酵素とα－グルコシダーゼによる分岐メガロ糖生成反応における枝切り酵素の影響
　シクロデキストリン生成酵素とα－グルコシダーゼによる分岐メガロ糖生成反応における枝切り酵素の影響を調べた。すなわち、シクロデキストリン生成酵素とα－グルコシダーゼに加えて、イソアミラーゼ、プルラナーゼあるいはこの両者をデンプン液化液に添加したものを未添加区と比較した。 2-2: Effect of debranching enzyme on the production of branched megalosaccharide by cyclodextrin-producing enzyme and α-glucosidase The influence of the debranching enzyme on the reaction of producing branched megalosaccharide by cyclodextrin-producing enzyme and α-glucosidase was investigated. That is, in addition to cyclodextrin-forming enzyme and α-glucosidase, isoamylase and / or pullulanase or both added to the starch liquefaction solution were compared with the non-added group.
　イソアミラーゼ添加区にはシュードモナス　アミロデラモサ由来のものを対固形分１ｇ当たり２００単位、プルラナーゼ添加区にはプルラナーゼ「アマノ」３、を対固形分当たり０．０２％添加した。シクロデキストリン生成酵素にはパエニバシルス　エスピー由来のものを用い、これをそれぞれ対固形分１ｇ当たり２単位添加した。また、α－グルコシダーゼにはアクレモニウム　エスピー由来のものを用い、これをそれぞれ対固形分１ｇ当たり０．６５単位添加した。 In the isoamylase-added section, Pseudomonas amyloderamosa-derived 200 units per gram of solid content was added, and in the pullulanase-added section, pullulanase “Amano” 3 was added at 0.02% per solid content. As the cyclodextrin-producing enzyme, one derived from Paenibacils sp. Was used, and 2 units were added per 1 g of the solid content. Further, α-glucosidase derived from Acremonium sp. Was used, and 0.65 units were added per gram of the solid content.
　基質には３０質量％ＤＥ６．５のコーンスターチ液化液を用い、この温度５３℃、ｐＨ６．０に調整し、上記各酵素をそれぞれ添加して７２時間糖化した。 30% by mass DE6.5 corn starch liquefied liquid was used as a substrate, the temperature was adjusted to 53 ° C. and pH 6.0, and each of the above enzymes was added for saccharification for 72 hours.
　得られた反応液を上記と同様にβ－アミラーゼ処理を行った。その結果を表２に示した。
Figure JPOXMLDOC01-appb-T000002
 （上記表中、表中の成分以外は重合度１０以下の低分子オリゴ糖であり、デキストリンは重合度３５を超える高分子成分を指す。） The resulting reaction solution was treated with β-amylase as described above. The results are shown in Table 2.
Figure JPOXMLDOC01-appb-T000002
 (In the above table, components other than the components in the table are low-molecular oligosaccharides having a degree of polymerization of 10 or less, and dextrin refers to a polymer component having a degree of polymerization of more than 35.)
　枝切り酵素未添加区では、重合度３５以上のデキストリン成分が反応後に残存した。一方、枝切り酵素添加区ではこのデキストリン成分の残存量が少なく、特にイソアミラーゼ添加区においてデキストリン成分の残存量が少なく、分岐メガロ糖成分が未添加区と比較して多かった。このことから、分岐メガロ糖の製造には枝切り酵素、特にイソアミラーゼの添加が有効であることが明らかになった。 In the section where no debranching enzyme was added, a dextrin component having a polymerization degree of 35 or more remained after the reaction. On the other hand, the amount of dextrin component remaining was small in the debranching enzyme-added group, and in particular, the amount of dextrin component remaining was small in the isoamylase-added group, and the branched megalosaccharide component was large compared to the non-added group. From this, it became clear that the addition of a debranching enzyme, especially isoamylase, is effective for the production of branched megalosaccharides.
試験例３：分岐メガロ糖の耐老化性の確認
　分岐メガロ糖を含有する液糖の液状品での保存安定性を調べることを目的として、分岐メガロ糖含有シラップを調製し、老化による白濁の有無により耐老化性を評価した。 Test Example 3: Confirmation of Aging Resistance of Branched Megalosaccharide For the purpose of investigating the storage stability of a liquid sugar containing a branched megalosaccharide in a liquid product, a syrup containing a branched megalosaccharide was prepared and checked for turbidity due to aging. Was used to evaluate aging resistance.
　３０質量％ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アクレモニウム　エスピーのα－グルコシダーゼを対固形分１ｇ当たり０．６５単位添加して７２時間糖化した。 30 mass% DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and pH 6.0, and to this, Paenibacils sp. Cyclodextrin-forming enzyme was 1 unit per gram of solid content, and Myloides odoratus isoamylase per gram of solid content. Saccharification was carried out for 72 hours by adding 100 units, 0.01% of pullulanase “Amano” 3 per solid and 0.65 units of Acremonium sp. Α-glucosidase per gram of solid.
　得られた糖化反応液（後記製造例５のシラップと同等）を８０℃で３０分間加熱処理を行い、α－アミラーゼ製剤である大和化成社製のクライスターゼＬ１を対固形分当たり０．００５％添加して、ヨード反応が消失するまで反応させた。これのｐＨを４．０として酵素を失活させた後、ろ過、イオン交換精製、活性炭処理して中性糖を精製した。イオン交換精製にはアンバーライトＭＢ３、活性炭処理には精製白鷺（キリンフードテック社）を使用した。精製後の糖液をＢｘ７５に濃縮して得られた液糖を室温に１ヶ月間保持した。得られた糖液の糖組成を表３に示した。 The resulting saccharification reaction solution (equivalent to syrup in Production Example 5 described later) was heated at 80 ° C. for 30 minutes, and α-amylase preparation Christase L1 manufactured by Daiwa Kasei Co., Ltd. was 0.005% per solid content. The reaction was continued until the iodine reaction disappeared. The enzyme was inactivated by adjusting the pH to 4.0, and then neutral sugar was purified by filtration, ion exchange purification, and activated carbon treatment. Amberlite MB3 was used for ion exchange purification, and purified white birch (Kirin Foodtech) was used for activated carbon treatment. The liquid sugar obtained by concentrating the purified sugar liquid to Bx75 was kept at room temperature for 1 month. Table 3 shows the sugar composition of the resulting sugar solution.
　デンプンのα－アミラーゼ分解物（コーンシラップ）を分岐メガロ糖含有シラップとの保存安定性の比較対照とした。すなわち、２２．５質量％ＤＥ６．５コーンスターチ液化液にクライスターゼＬ１を対固形分当たり０．００５％、プルラナーゼ「アマノ」３を対固形分当たり０．０４％、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり４００単位添加して３６時間ｐＨ６．０、５３℃で３６時間糖化した。これを８０℃に加温した後、クライスターゼＬ１を対固形分当たり０．００５％追添加してヨード反応が消失するまで保持した。得られた反応液より中性糖を上記と同様に精製、濃縮した。得られた糖液の糖組成を表３に示した。
Figure JPOXMLDOC01-appb-T000003
 （上記表中、表中の成分以外は重合度１０以下の低分子オリゴ糖である。メガロ糖は重合度１１～３５の糖質を指し、カッコ内は分岐メガロ糖含量を示す。デキストリンは重合度３５を超える高分子成分を指す。） An α-amylase degradation product (corn syrup) of starch was used as a comparative control of storage stability with branched megalosaccharide-containing syrup. That is, 22.5% by mass of DE 6.5 corn starch liquefied liquid was obtained by adding 0.005% of the Christase L1 to the solid content, 0.04% of the pullulanase “Amano” 3 to the solid content, and Myloides odolatas isoamylase to the solid. 400 units were added per gram per minute and saccharified for 36 hours at pH 6.0 and 53 ° C. After heating this to 80 ° C., Christase L1 was additionally added to the solid content by 0.005% and held until the iodine reaction disappeared. Neutral sugars were purified and concentrated in the same manner as above from the resulting reaction solution. Table 3 shows the sugar composition of the resulting sugar solution.
Figure JPOXMLDOC01-appb-T000003
 (In the above table, components other than the components in the table are low-molecular oligosaccharides having a polymerization degree of 10 or less. Megalosaccharide refers to a saccharide having a polymerization degree of 11 to 35, and the content in parentheses indicates branched megalosaccharide content. Refers to a polymer component having a degree exceeding 35.)
　また、図２に示すように、コーンシラップでは老化のため、保存後１ヶ月で白色沈澱を生じ、糖液が白濁した。しかし、分岐メガロ糖含有シラップはＤＰ１０～３５程度の糖質の含有量が、コーンシラップと比較して高いにも関わらず、いずれも保存１ヵ月後に白濁を生じなかった。 In addition, as shown in FIG. 2, corn syrup was aged, resulting in white precipitation in one month after storage, and the sugar solution became cloudy. However, none of the branched megalosaccharide-containing syrup had white turbidity after 1 month of storage, although the content of carbohydrates of DP 10 to 35 was higher than that of corn syrup.
　以上の結果から、分岐メガロ糖は耐老化性に優れ、液状品としての保存安定性が高いことが明らかになった。 From the above results, it was clarified that branched megalosaccharide has excellent aging resistance and high storage stability as a liquid product.
試験例４：分岐メガロ糖含有シラップの粘度の測定
　試験例３で製造した分岐メガロ糖含有シラップ（製造例５のシラップと同等）の粘度を測定し、既存オリゴ糖シラップと比較した。すなわち、試験例３で調製した分岐メガロ糖含有シラップの１０℃～６０℃における粘度を測定した。シラップの濃度をＢｘ７０とした。粘度の測定には東機産業社製ＶＩＳＣＯＭＥＴＥＲ　ＴＶＢ－１０を用い、ローターの回転数を６０ｒｐｍとし、測定時間を４０秒間とした。サンプル３００ｍＬを３００ｍＬ容トールビーカーに入れ、測定サンプルとした。各温度における分岐メガロ糖含有シラップの粘度は、既存高分子マルトオリゴ糖含有シラップである日本食品化工社製フジオリゴＧ６７と比較して高かったものの、固形分７０％においても操作性に問題ない程度の粘度であった。 Test Example 4: Measurement of Viscosity of Branched Megalosaccharide-Containing Syrup The viscosity of the branched megalosaccharide-containing syrup produced in Test Example 3 (equivalent to the syrup of Production Example 5) was measured and compared with the existing oligosaccharide syrup. That is, the viscosity at 10 ° C. to 60 ° C. of the branched megalosaccharide-containing syrup prepared in Test Example 3 was measured. The concentration of syrup was Bx70. The viscosity was measured using VISCOMETER TVB-10 manufactured by Toki Sangyo Co., Ltd., the rotation speed of the rotor was 60 rpm, and the measurement time was 40 seconds. A 300 mL sample was placed in a 300 mL tall beaker and used as a measurement sample. Although the viscosity of the branched megalosaccharide-containing syrup at each temperature was higher than that of Fuji Oligo G67 manufactured by Nippon Shokuhin Kako Co., Ltd., which is an existing polymer maltooligosaccharide-containing syrup, the viscosity is such that there is no problem in operability even at a solid content of 70%. Met.
試験例５：分岐メガロ糖含有シラップの甘味度の測定
　試験例３で製造した分岐メガロ糖含有シラップ（製造例５のシラップと同等）の甘味度をＰａｕｌｉの全系列法（澱粉糖関連工業分析法　澱粉糖技術部会編　６１～６２頁）により測定した。すなわち、０．６～１．４％の範囲、０．２％間隔で調製した砂糖水溶液と固形分１０％の分岐メガロ糖含有シラップの甘味度を「強」、「等」、「弱」で１０人のパネラーに評価させた。得られた試験結果を基に、次式により甘味度を算出した。
Ｅｏ＝（Ｄｏ＋ｉ／２）－Σｇｒ・ｉ／ｎ
Ｅｕ＝（Ｄｕ－ｉ／２）＋Σｋｌ・ｉ／ｎ
Ｅｍ＝（Ｅｏ＋Ｅｕ）／２
Ｓ＝Ｅｍ／Ｎ・１００
Ｄｏ＝標準糖液の最も低い濃度；　Ｄｕ＝標準糖液の最も高い濃度；　ｉ＝標準糖液の濃度間隔；　Σｇｒ＝「強」と判定した人数；　Σｋｌ＝「弱」と判定した人数；　ｎ＝パネラー数；　Ｓ＝甘味度；　Ｎ＝試料濃度（％） Test Example 5: Measurement of Sweetness of Branched Megalosaccharide-Containing Syrup The sweetness of the branched megalosaccharide-containing syrup produced in Test Example 3 (equivalent to the syrup of Production Example 5) was measured by Pauli's whole series method (starch sugar related industrial analysis method) Measured according to Starch Sugar Technical Committee, pages 61-62). That is, the sweetness of the sugar aqueous solution prepared at intervals of 0.2% in the range of 0.6 to 1.4% and the branched megalosaccharide-containing syrup having a solid content of 10% is “strong”, “equal”, “weak”. Ten panelists evaluated. Based on the obtained test results, the sweetness degree was calculated by the following formula.
Eo = (Do + i / 2) −Σgr · i / n
Eu = (Du−i / 2) + Σkl · i / n
Em = (Eo + Eu) / 2
S = Em / N · 100
Do = lowest concentration of standard sugar solution; Du = highest concentration of standard sugar solution; i = concentration interval of standard sugar solution; Σgr = number of people judged as “strong”; Σkl = number of people judged as “weak”; n = Number of panelists; S = sweetness; N = sample concentration (%)
　その結果、分岐メガロ糖含有シラップの甘味度は１０．７となり、既存の液糖と比較して極めて甘味度が低いことが明らかになった。 As a result, the sweetness of the branched megalosaccharide-containing syrup was 10.7, and it was revealed that the sweetness was extremely low compared with the existing liquid sugar.
製造例１：分岐メガロ糖の製造（１）
（１）分岐メガロ糖の製造
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アスペルギルス　ニガーのα－グルコシダーゼを対固形分１ｇ当たり３．７５単位添加して７２時間糖化した。これを８０℃に加温し、クライスターゼＬ１を対固形分当たり０．００５％添加してヨード反応が消失するまで作用させた。このｐＨを４．０として酵素を失活させたものに、活性炭を添加して常法にしたがって脱色し、珪藻土によりろ過してろ液を得た。これを常法にしたがイオン交換樹脂によりイオン性成分を除去し、中性糖を得た。これに上記と同様に再度活性炭処理を行うことにより脱色し、珪藻土により濾過してろ液を得た。これを減圧下で固形分７５％となるまで濃縮した。以上の操作により、分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり２０．３％含有しており、β－アミラーゼ処理により分岐メガロ糖含有量を測定したところ、重合度１１～３５の分岐メガロ糖を対固形分当たり１７．９％含有していた。 Production Example 1: Production of branched megalosaccharide (1)
(1) Production of branched megalosaccharide 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Saccharification for 72 hours by adding 100 units per gram of odoratus isoamylase, 0.01% of pullulanase “Amano” 3 per solid, and 3.75 units of α-glucosidase of Aspergillus niger per gram of solid did. This was heated to 80 ° C., and Christase L1 was added in an amount of 0.005% to the solid content until the iodine reaction disappeared. Activated charcoal was added to the inactivated enzyme at pH 4.0 and decolorized according to a conventional method, followed by filtration through diatomaceous earth to obtain a filtrate. Although this was made into a usual method, the ionic component was removed with the ion exchange resin, and the neutral sugar was obtained. This was decolorized by performing the activated carbon treatment again in the same manner as above, and filtered through diatomaceous earth to obtain a filtrate. This was concentrated under reduced pressure to 75% solids. By the above operation, a branched megalosaccharide-containing syrup was obtained at a yield of about 90% based on the solid content. This product contains 20.3% of megalosaccharide per solid, and when the content of branched megalosaccharide was measured by β-amylase treatment, branched megalosaccharide having a degree of polymerization of 11 to 35 per solid was measured. It contained 17.9%.
（２）分岐メガロ糖の分岐構造の解析１
　上記（１）においてアスペルギルス　ニガーを使用して得られた分岐メガロ糖の分岐構造の結合様式を解析した。すなわち、上記（１）において得られた糖液を凍結乾燥後、粉末状にし、得られた粉末５ｍｇを減圧下１０５℃で数時間保持することにより乾燥し、これにＤＭＳＯ０．５ｍＬを添加して溶解した。これに微粉砕したＮａＯＨ６０ｍｇを添加して時々攪拌しながら室温に１時間保持した。次いでヨードメタン０．３ｍＬを添加し、室温に１時間保持した後に６０℃に１時間保持した。これに水１ｍＬを添加して反応を停止し、クロロホルム１ｍＬを添加して攪拌した。水層を除去し、新たに水１ｍＬを添加、攪拌する操作を５回繰り返し、クロロホルム層を回収した。次いで、無水Ｎａ２ＳＯ４を添加して乾燥させ、６０℃にてクロロホルムを蒸発させた。これに４Ｍトリフルオロ酢酸１ｍＬを加えて１００℃に４時間保持した。これをナス型フラスコに移してロータリーエバポレーターにて乾固させ、水０．５ｍＬを加えて溶解し、アンモニア水を３滴加えてアルカリ性にした。次いでホウ素化水素化ナトリウム１０ｍｇを添加して室温に一晩保持した。これに酢酸を加えて反応を停止し、メタノールを加えて乾固させた。メタノールを添加して乾固させる操作を５回繰り返し、ピリジンおよび無水酢酸をそれぞれ２ｍＬずつ添加して１００℃に４時間保持した。これに水を加えて反応を停止し、クロロホルムにより抽出を行った。水層を除去し、新たに水１ｍＬを添加、攪拌する操作を５回繰り返し、クロロホルム層を回収した。得られた試料をガスクロマトグラフィーに供し、部分メチル化単糖のアルジトールアセタートの解析を行った。ガスクロマトグラフィーではカラムにＴＣ－１７（０．２５ｍｍｘ３０ｍ；ＧＬサイエンス製）を用い、カラム温度を５０℃に１分間保持した後、１分間当たり１０℃ずつカラム温度を２８０℃まで上昇させた。サンプル注入温度を３００℃とした。検出をＦＩＤにより行い、検出温度を３００℃とした。キャリアーガスにはヘリウムを使用し、流速を２．５ｍＬ／分とした。 (2) Analysis of branched structure of branched megalosaccharide 1
The binding mode of the branched structure of the branched megalosaccharide obtained by using Aspergillus niger in the above (1) was analyzed. That is, the sugar solution obtained in the above (1) was freeze-dried and then powdered, and 5 mg of the obtained powder was dried by holding at 105 ° C. under reduced pressure for several hours, to which 0.5 mL of DMSO was added. Dissolved. To this, 60 mg of finely pulverized NaOH was added and kept at room temperature for 1 hour with occasional stirring. Subsequently, 0.3 mL of iodomethane was added, and it hold | maintained at 60 degreeC for 1 hour, after hold | maintaining at room temperature for 1 hour. 1 mL of water was added thereto to stop the reaction, and 1 mL of chloroform was added and stirred. The aqueous layer was removed, and 1 mL of water was newly added and stirred for 5 times, and the chloroform layer was recovered. Then, anhydrous Na 2 SO 4 was added and dried, and chloroform was evaporated at 60 ° C. To this, 1 mL of 4M trifluoroacetic acid was added and kept at 100 ° C. for 4 hours. This was transferred to an eggplant-shaped flask, dried on a rotary evaporator, dissolved by adding 0.5 mL of water, and made alkaline by adding 3 drops of aqueous ammonia. Then 10 mg of sodium borohydride was added and kept at room temperature overnight. Acetic acid was added thereto to stop the reaction, and methanol was added to dryness. The operation of adding methanol to dryness was repeated 5 times, and 2 mL each of pyridine and acetic anhydride was added and kept at 100 ° C. for 4 hours. Water was added thereto to stop the reaction, and extraction was performed with chloroform. The aqueous layer was removed, and 1 mL of water was newly added and stirred for 5 times, and the chloroform layer was recovered. The obtained sample was subjected to gas chromatography to analyze the partially methylated monosaccharide alditol acetate. In the gas chromatography, TC-17 (0.25 mm × 30 m; manufactured by GL Science) was used for the column, the column temperature was kept at 50 ° C. for 1 minute, and then the column temperature was increased to 280 ° C. by 10 ° C. per minute. The sample injection temperature was 300 ° C. Detection was performed by FID, and the detection temperature was 300 ° C. Helium was used as the carrier gas, and the flow rate was 2.5 mL / min.
　その結果、アスペルギルス　ニガー由来のα－グルコシダーゼを使用して得られた分岐メガロ糖に由来する部分メチル化誘導体として、１，５－ジ－Ｏ－アセチル－２，３，４，６－テトラ－Ｏ－メチルヘキシトール、１，５，６－トリ－Ｏ－アセチル－２，３，４－トリ－Ｏ－メチルヘキシトールおよび１，４，５－トリ－Ｏ－アセチル－２，３，６，－トリ－Ｏ－メチルヘキシトールが検出された。このことから、製造例１（１）で得られた分岐メガロ糖の分岐構造はα－１，６－結合であることが明らかになった。 As a result, 1,5-di-O-acetyl-2,3,4,6-tetra-O was obtained as a partially methylated derivative derived from branched megalosaccharide obtained using α-glucosidase derived from Aspergillus niger. -Methylhexitol, 1,5,6-tri-O-acetyl-2,3,4-tri-O-methylhexitol and 1,4,5-tri-O-acetyl-2,3,6 , -Tri-O-methylhexitol was detected. This revealed that the branched structure of the branched megalosaccharide obtained in Production Example 1 (1) is an α-1,6-bond.
（３）分岐メガロ糖の分岐構造の解析２
　上記（１）において得られた分岐メガロ糖を１０％（ｗ／ｖ）となるように調整し、この水溶液１Ｌに１Ｍ酢酸緩衝液（ｐＨ５．５）を５ｍｌ、アマノエンザイム社製ＡＭＴ　１．２Ｌを４．６ｍｌおよびナガセケムテックス社製ベータ－アミラーゼ＃１５００を０．１ｇ添加し、５３℃にて５時間反応させた。この反応により、直鎖糖を３糖以下に低分子化し、また、分岐メガロ糖の還元末端の直鎖部分を加水分解することにより、非還元末端部分を含む糖をオリゴ糖化した。得られた反応液を０．４５μｍフィルターによりろ過したものを濃縮し、分画原資とした。これをカーボン－セライトカラムクロマトグラフィーに供した。本クロマトグラフィーでは０－３％のｎ－ブタノールの直線濃度勾配により糖を溶出させた。得られた５糖画分をそれぞれイオン交換樹脂にて脱塩後、０．４５μｍフィルターによりろ過したものを凍結乾燥した。このうち５ｍｇを上記と同様にメチル化分析に供した。 (3) Analysis of branched structure of branched megalosaccharide 2
The branched megalosaccharide obtained in (1) above was adjusted to 10% (w / v), 5 ml of 1M acetate buffer (pH 5.5) was added to 1 L of this aqueous solution, and AMT 1.2 L manufactured by Amano Enzyme. And 0.1 g of Nagase ChemteX Beta-Amylase # 1500 were added and reacted at 53 ° C. for 5 hours. By this reaction, the saccharide containing the non-reducing end portion was oligosaccharide-ized by reducing the molecular weight of the linear saccharide to 3 sugars or less and hydrolyzing the reducing end linear portion of the branched megalosaccharide. The reaction solution obtained by filtration through a 0.45 μm filter was concentrated and used as a fraction source. This was subjected to carbon-celite column chromatography. In this chromatography, sugar was eluted with a linear concentration gradient of 0-3% n-butanol. The obtained pentasaccharide fraction was desalted with an ion exchange resin, and then filtered through a 0.45 μm filter and freeze-dried. Of this, 5 mg was subjected to methylation analysis as described above.
　その結果、製造例１（１）で得られた分岐メガロ糖に由来する分岐５糖（５糖画分）では、還元末端側４残基に由来する１，５，６－トリ－Ｏ－アセチル－２，３，４－トリ－Ｏ－メチルヘキシトールおよび１，４，５－トリ－Ｏ－アセチル－２，３，６－トリ－Ｏ－メチルヘキシトールのピーク面積の比が１：３となった。これは本分岐糖の還元末端側のグルコシル基の結合比を示しており、非還元末端側の分岐構造が平均してグルコシル基１残基からなることを意味している。 As a result, in the branched pentasaccharide (pentasaccharide fraction) derived from the branched megalosaccharide obtained in Production Example 1 (1), 1,5,6-tri-O-acetyl derived from four residues on the reducing end side. The ratio of the peak areas of -2,3,4-tri-O-methylhexitol and 1,4,5-tri-O-acetyl-2,3,6-tri-O-methylhexitol is 1: It became 3. This shows the binding ratio of the glucosyl group on the reducing end side of the branched sugar, and means that the branched structure on the non-reducing end side consists of one residue of glucosyl group on average.
製造例２：分岐メガロ糖の製造（２）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにバチルス　コアギュランスのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アスペルギルス　ニガーのα－グルコシダーゼを対固形分１ｇ当たり３．７５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり１７．５％含有しており、分岐メガロ糖を対固形分当たり１５．９％含有していた。 Production Example 2: Production of branched megalosaccharide (2)
A 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus coagulans cyclodextrin-producing enzyme was paired with 1 unit per gram of solid content and Myloides odoratus isoamylase. Saccharification was performed for 60 hours by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 3.75 units of α-glucosidase of Aspergillus niger per gram of solid content. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. In addition, this product contained 17.5% of megalo sugar per solid and 15.9% of branched megalo sugar per solid.
製造例３：分岐メガロ糖の製造（３）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにバチルス　ステアロサーモフィルスのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アスペルギルス　ニガーのα－グルコシダーゼを対固形分１ｇ当たり３．７５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり１８．３％含有しており、分岐メガロ糖を対固形分当たり１５．５％含有していた。 Production Example 3: Production of branched megalosaccharide (3)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus stearothermophilus cyclodextrin-forming enzyme was added to 1 unit per gram of solid content, and Myloides odoratus isoform. Saccharification was performed by adding 100 units per gram of amylase, 0.01% of pullulanase “Amano” 3 per solid, and 3.75 units of α-glucosidase of Aspergillus niger per gram of solid for 60 hours. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. This product contained 18.3% of megalosaccharide per solid and 15.5% of branched megalosaccharide per solid.
製造例４：分岐メガロ糖の製造（４）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにバチルス　マゼランスのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アスペルギルス　ニガーのα－グルコシダーゼを対固形分１ｇ当たり３．７５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり１５．３％含有しており、分岐メガロ糖を対固形分当たり１４．３％含有していた。 Production Example 4: Production of branched megalosaccharide (4)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus mazellans cyclodextrin-forming enzyme was paired with 1 unit per gram of solids and Myloides odoratus isoamylase. Saccharification was performed for 60 hours by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 3.75 units of α-glucosidase of Aspergillus niger per gram of solid content. The subsequent operation was carried out in the same manner as in Production Example 1, and a branched megalosaccharide-containing syrup having a solid content of 75% was obtained at a yield of about 90% per solid content. In addition, this product contained 15.3% of megalo sugar per solid, and contained 14.3% of branched megalo sugar per solid.
製造例５：分岐メガロ糖の製造（５）
（１）分岐メガロ糖の製造
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アクレモニウム　エスピーのα－グルコシダーゼを対固形分１ｇ当たり０．６５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり８１．３％含有しており、分岐メガロ糖を対固形分当たり５６．１％含有していた。 Production Example 5: Production of branched megalosaccharide (5)
(1) Manufacture of branched megalosaccharide 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. 100 mg / g of Myloides odoratus isoamylase, 0.01% of pullulanase “Amano” 3 per g of solid, and 0.65 unit of α-glucosidase of Acremonium sp. Saccharified for 60 hours. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. The product contained 81.3% of megalosaccharide per solid and 56.1% of branched megalosaccharide per solid.
（２）分岐メガロ糖の分岐構造の解析１
　（１）においてアクレモニウム　エスピー由来のα－グルコシダーゼを使用して得られた分岐メガロ糖の分岐構造の結合様式を解析した。分岐メガロ糖の調製及びメチル化分析は、製造例１（２）と同様に行った。 (2) Analysis of branched structure of branched megalosaccharide 1
In (1), the binding mode of the branched structure of the branched megalosaccharide obtained by using α-glucosidase derived from Acremonium sp. Was analyzed. Preparation of branched megalosaccharide and methylation analysis were performed in the same manner as in Production Example 1 (2).
　その結果、アクレモニウム　エスピー由来のα－グルコシダーゼを使用して得られた分岐メガロ糖に由来する部分メチル化誘導体として、１，５－ジ－Ｏ－アセチル－２，３，４，６－テトラ－Ｏ－メチルヘキシトール、１，３，５－トリ－Ｏ－アセチル－２，４，６－トリ－Ｏ－メチルヘキシトールおよび１，４，５－トリ－Ｏ－アセチル－２，３，６－トリ－Ｏ－メチルヘキシトールが検出された。このことから、製造例５（１）で得られた分岐メガロ糖の分岐構造はα－１，３－結合であることが明らかになった。 As a result, 1,5-di-O-acetyl-2,3,4,6-tetra- was obtained as a partially methylated derivative derived from branched megalosaccharide obtained by using α-glucosidase derived from Acremonium sp. O-methylhexitol, 1,3,5-tri-O-acetyl-2,4,6-tri-O-methylhexitol and 1,4,5-tri-O-acetyl-2,3 6-Tri-O-methylhexitol was detected. This revealed that the branched structure of the branched megalosaccharide obtained in Production Example 5 (1) is an α-1,3-linkage.
（３）分岐メガロ糖の分岐構造の解析２
　上記（１）において得られた分岐メガロ糖を１０％（ｗ／ｖ）となるように調整し、この水溶液１Ｌに１Ｍ酢酸緩衝液（ｐＨ５．５）を５ｍｌ、アマノエンザイム社製ＡＭＴ　１．２Ｌを４．６ｍｌおよびナガセケムテックス社製ベータ－アミラーゼ＃１５００を０．１ｇ添加し、５３℃にて５時間反応させた。この反応により、直鎖糖を３糖以下に低分子化し、また、分岐メガロ糖の還元末端の直鎖部分を加水分解することにより、非還元末端部分を含む糖をオリゴ糖化した。得られた反応液を０．４５μｍフィルターによりろ過したものを濃縮し、分画原資とした。これをカーボン－セライトカラムクロマトグラフィーに供した。本クロマトグラフィーでは０－３％のｎ－ブタノールの直線濃度勾配により糖を溶出させた。得られた５糖画分をそれぞれイオン交換樹脂にて脱塩後、０．４５μｍフィルターによりろ過したものを凍結乾燥した。このうち５ｍｇを上記と同様にメチル化分析に供した。 (3) Analysis of branched structure of branched megalosaccharide 2
The branched megalosaccharide obtained in (1) above was adjusted to 10% (w / v), 5 ml of 1M acetate buffer (pH 5.5) was added to 1 L of this aqueous solution, and AMT 1.2 L manufactured by Amano Enzyme. And 0.1 g of Nagase ChemteX Beta-Amylase # 1500 were added and reacted at 53 ° C. for 5 hours. By this reaction, the saccharide containing the non-reducing end portion was oligosaccharide-ized by reducing the molecular weight of the linear saccharide to 3 sugars or less and hydrolyzing the reducing end linear portion of the branched megalosaccharide. The reaction solution obtained by filtration through a 0.45 μm filter was concentrated and used as a fraction source. This was subjected to carbon-celite column chromatography. In this chromatography, sugar was eluted with a linear concentration gradient of 0-3% n-butanol. The obtained pentasaccharide fraction was desalted with an ion exchange resin, and then filtered through a 0.45 μm filter and freeze-dried. Of this, 5 mg was subjected to methylation analysis as described above.
　その結果、製造例５（１）で得られた分岐メガロ糖に由来する分岐５糖（５糖画分）では、還元末端側４残基からの１，３，５－トリ－Ｏ－アセチル－２，４，６－トリ－Ｏ－メチルヘキシトールおよび１，４，５－トリ－Ｏ－アセチル－２，３，６－トリ－Ｏ－メチルヘキシトールのピーク面積比が１：１となった。このことから、非還元末端側の分岐構造が平均してグルコシル基２残基からなると推定された。 As a result, in the branched pentasaccharide (pentasaccharide fraction) derived from the branched megalosaccharide obtained in Production Example 5 (1), 1,3,5-tri-O-acetyl-from the reducing end side 4 residues was obtained. The peak area ratio of 2,4,6-tri-O-methylhexitol and 1,4,5-tri-O-acetyl-2,3,6-tri-O-methylhexitol is 1: 1. became. From this, it was estimated that the branched structure on the non-reducing terminal side was composed of 2 residues of glucosyl groups on average.
製造例６：分岐メガロ糖の製造（６）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにバチルス　コアギュランスのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アクレモニウム　エスピーのα－グルコシダーゼを対固形分１ｇ当たり０．６５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり７７．５％含有しており、分岐メガロ糖を対固形分当たり５１．２％含有していた。 Production Example 6: Production of branched megalosaccharide (6)
A 30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus coagulans cyclodextrin-producing enzyme was paired with 1 unit per gram of solid content and Myloides odoratus isoamylase. Saccharification was carried out by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 0.65 units of Acremonium sp. Α-glucosidase per gram of solid content for 60 hours. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. This product contained 77.5% megalosaccharide per solid and 51.2% branched megalosaccharide per solid.
製造例７：分岐メガロ糖の製造（７）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにバチルス　ステアロサーモフィルスのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アクレモニウム　エスピーのα－グルコシダーゼを対固形分１ｇ当たり０．６５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり６６．２％含有しており、分岐メガロ糖を対固形分当たり５４．８％含有していた。 Production Example 7: Production of branched megalosaccharide (7)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus stearothermophilus cyclodextrin-forming enzyme was added to 1 unit per gram of solid content, and Myloides odoratus isoform. Saccharification was performed by adding 100 units per gram of amylase, 0.01% of pullulanase “Amano” 3 per solid, and 0.65 units of Acremonium sp. Α-glucosidase per gram of solid for 60 hours. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. In addition, this product contained 66.2% of megalo sugar per solid and 54.8% of branched megalo sugar per solid.
製造例８：分岐メガロ糖の製造（８）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにバチルス　マゼランスのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、マイロイデス　オドラータスのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アクレモニウム　エスピーのα－グルコシダーゼを対固形分１ｇ当たり０．６５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり５４．０％含有しており、分岐メガロ糖を対固形分当たり４７．７％含有していた。 Production Example 8: Production of branched megalosaccharide (8)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Bacillus mazellans cyclodextrin-forming enzyme was paired with 1 unit per gram of solids and Myloides odoratus isoamylase. Saccharification was carried out by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 0.65 units of Acremonium sp. Α-glucosidase per gram of solid content for 60 hours. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. In addition, this product contained 54.0% of megalo sugar per solid and 47.7% of branched megalo sugar per solid.
製造例９：分岐メガロ糖の製造（９）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形分１ｇ当たり２単位、シュードモナス　アミロデラモサのイソアミラーゼを対固形分１ｇ当たり２００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０２％、アスペルギルス　ニガーのα－グルコシダーゼを対固形分１ｇ当たり３．７５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり１６．９％含有しており、分岐メガロ糖を対固形分当たり１６．４％含有していた。 Production Example 9: Production of branched megalosaccharide (9)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and pH 6.0, to which Paenibacillus sp. Cyclodextrin-producing enzyme was paired with 2 units per gram of solid content and Pseudomonas amylodelamosa isoamylase. Saccharification was carried out for 60 hours by adding 200 units per gram of solid content, 0.02% of pullulanase “Amano” 3 per solid content, and 3.75 units of α-glucosidase of Aspergillus niger per gram of solid content. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. In addition, this product contained 16.9% of megalo sugar per solid and 16.4% of branched megalo sugar per solid.
製造例１０：分岐メガロ糖の製造（１０）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、シュードモナス　アミロデラモサのイソアミラーゼを対固形分１ｇ当たり１００単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アクレモニウム　エスピーのα－グルコシダーゼを対固形分１ｇ当たり０．６５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり５３．４％含有しており、分岐メガロ糖を対固形分当たり５０．８％含有していた。 Production Example 10: Production of branched megalosaccharide (10)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-producing enzyme was paired with 1 unit per gram of solid content and Pseudomonas amylodelamosa isoamylase was paired. Saccharification was carried out by adding 100 units per gram of solid content, 0.01% of pullulanase “Amano” 3 per solid content, and 0.65 units of Acremonium sp. Α-glucosidase per gram of solid content for 60 hours. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. In addition, this product contained 53.4% of megalo sugar per solid and 50.8% of branched megalo sugar per solid.
製造例１１：分岐メガロ糖の製造（１１）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アスペルギルス　ニガーのα－グルコシダーゼを対固形分１ｇ当たり３．７５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり１６．０％含有しており、分岐メガロ糖を対固形分当たり１１．７％含有していた。 Production Example 11 Production of Branched Megalosaccharide (11)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-forming enzyme was paired with 1 unit per 1 g of solid content and pullulanase “Amano” 3 was paired. 0.01% per solid, Aspergillus niger α-glucosidase was added to 3.75 units per gram of solid and saccharified for 60 hours. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. In addition, this product contained 16.0% of megalosaccharide per solid and 11.7% of branched megalosaccharide per solid.
製造例１２：分岐メガロ糖の製造（１２）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、プルラナーゼ「アマノ」３を対固形分当たり０．０１％、アクレモニウム　エスピーのα－グルコシダーゼを対固形分１ｇ当たり０．６５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり３７．９％含有しており、分岐メガロ糖を対固形分当たり３３．３％含有していた。 Production Example 12: Production of branched megalosaccharide (12)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-producing enzyme was paired with 1 unit per 1 g of solid content, and pullulanase “Amano” 3 was paired. Saccharification was carried out for 60 hours by adding 0.65 unit of α-glucosidase of 0.01% per solid and 0.01% Acremonium sp. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. In addition, this product contained 37.9% of megalo sugar per solid content, and contained 33.3% of branched megalo sugar per solid content.
製造例１３：分岐メガロ糖の製造（１３）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、アスペルギルス　ニガーのα－グルコシダーゼを対固形分１ｇ当たり３．７５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり１７．３％含有しており、分岐メガロ糖を対固形分当たり１７．０％含有していた。 Production Example 13: Production of branched megalosaccharide (13)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-forming enzyme was added to 1 unit per gram of solid and Aspergillus niger α-glucosidase. Saccharification was carried out for 60 hours by adding 3.75 units per gram of the solid content. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. The product contained 17.3% of megalosaccharide per solid and 17.0% of branched megalosaccharide per solid.
製造例１４：分岐メガロ糖の製造（１４）
　３０％（ｗ／ｗ）ＤＥ６．５コーンスターチ液化液を温度５３℃、ｐＨ６．０に調整し、これにパエニバシルス　エスピーのシクロデキストリン生成酵素を対固形分１ｇ当たり１単位、アクレモニウム　エスピーのα－グルコシダーゼを対固形分１ｇ当たり０．６５単位添加して６０時間糖化した。以後の操作を製造例１と同様に行い、固形分７５％の分岐メガロ糖含有シラップを対固形分当たり約９０％の収率で得た。なお、本品はメガロ糖を対固形分当たり３２．６％含有しており、分岐メガロ糖を対固形分当たり２９．２％含有していた。 Production Example 14 Production of Branched Megalosaccharide (14)
30% (w / w) DE 6.5 corn starch liquefied liquid was adjusted to a temperature of 53 ° C. and a pH of 6.0, to which Paenibacillus sp. Cyclodextrin-producing enzyme was 1 unit per gram of solid content, Acremonium sp α-glucosidase Was saccharified for 60 hours by adding 0.65 units per gram of the solid content. The subsequent operation was carried out in the same manner as in Production Example 1 to obtain a branched megalosaccharide-containing syrup having a solid content of 75% in a yield of about 90% relative to the solid content. In addition, this product contained 32.6% of megalo sugar per solid content, and contained 29.2% of branched megalo sugar per solid content.
製造例１５：分岐メガロ糖含有粉飴の製造（１）
　製造例１により得られた糖液をＬ－１２型スプレードライヤー（アトマイザー式：大河原化工機）を使用して粉末化した。スプレードライヤーの入口設定温度を１６０℃、出口設定温度を１２０℃とし、アトマイザー回転数を１０，０００ｒｐｍ、サイクロン差圧を７０ｍｍＨｇ程度、給液量を５Ｌ／ｈとして運転した。この操作により、回収率約９５％で粉末状の分岐メガロ糖含有粉飴を得た。 Production Example 15: Production of powdered koji containing branched megalo sugar (1)
The sugar solution obtained in Production Example 1 was pulverized using an L-12 type spray dryer (atomizer type: Okawara Chemical). The spray dryer was operated at an inlet set temperature of 160 ° C., an outlet set temperature of 120 ° C., an atomizer rotational speed of 10,000 rpm, a cyclone differential pressure of about 70 mmHg, and a liquid supply amount of 5 L / h. By this operation, a powdered branched megalosaccharide-containing powder cake with a recovery rate of about 95% was obtained.
製造例１６：分岐メガロ糖含有粉飴の製造（２）
　製造例５により得られた糖液をＬ－１２型スプレードライヤーを用いて製造例１５と同様に噴霧乾燥した。この操作により、回収率約９５％で粉末状の分岐メガロ糖含有粉飴を得た。 Production Example 16: Production of branched megalosaccharide-containing flour (2)
The sugar solution obtained in Production Example 5 was spray-dried in the same manner as in Production Example 15 using an L-12 type spray dryer. By this operation, a powdered branched megalosaccharide-containing powder cake with a recovery rate of about 95% was obtained.
製造例１７：分岐メガロ糖分画品の製造
　製造例１の分岐メガロ糖含有シラップをＢｒｉｘ　５０程度に調製し、６０℃に加温したゲルろ過カラム（ＴＯＹＯＰＥＡＲＬ　ＨＷ－４０Ｓ，φ５．０ｃｍ　ｘ　８７ｃｍ，
　１７００ｍｌ）に供し、精製水を用い、流速を３４～４８ｍｌ／ｈに設定して溶出を行なった。溶出液の糖組成を高速液体クロマトグラフィーにより分析し、四糖以下の分岐糖含量が０．５％未満の画分（高分子画分）と０．５％以上の画分（低分子含有画分）をそれぞれ回収した。回収した画分を減圧乾燥によりＢｒｉｘ　３０程度まで濃縮した。以上の操作を１１回繰り返すことにより、固形分換算で高分子画分を２３ｇ、低分子含有画分を３９ｇ得た。 Production Example 17 Production of Branched Megalosaccharide Fraction Product A gel filtration column (TOYOPEARL HW-40S, φ5.0 cm × 87 cm, prepared by preparing the branched megalosaccharide-containing syrup of Production Example 1 to about Brix 50 and heating to 60 ° C.
Elution was carried out using purified water at a flow rate of 34 to 48 ml / h. The saccharide composition of the eluate was analyzed by high performance liquid chromatography, and the fraction of branched sugars below tetrasaccharide (high molecular fraction) and the fraction of 0.5% or higher (low molecular content fraction). Min) was collected. The collected fraction was concentrated to about Brix 30 by drying under reduced pressure. By repeating the above operation 11 times, 23 g of a polymer fraction and 39 g of a low-molecule-containing fraction were obtained in terms of solid content.
製造例１８：分岐メガロ糖還元物の製造
　分岐メガロ糖還元物をＮａＢＨ４を用いて調製した。すなわち、Ｂｘ１０に調整した分岐メガロ糖（製造例５）水溶液５ｍｌを氷中で１０分間冷却後、約２０ｍｇずつＮａＢＨ４を１０回添加して溶解した。この溶液にアンモニア水を２滴加え、ｐＨ１０以上とした後、室温にて３時間還元反応を行った。反応後、残存するＮａＢＨ４をＡｍｂｅｒｌｉｔｅ　ＭＢ－４（オルガノ）を水素の発生と樹脂の色調変化がなくなるまで添加して分解した。この液層を樹脂を水洗しながら回収し、ロータリーエバポレーターを用いて乾燥させた。これに１ｍｌメタノールを添加して溶解し、再度乾固させた。この操作を５回繰り返した。最後に精製水に溶解したものを還元物溶液とした。 Production Example 18 Production of Reduced Branched Megalosaccharide A branched megalosaccharide reduced product was prepared using NaBH 4 . That is, after cooling 10 minutes branched cytomegalovirus sugars adjusted to Bx10 (Preparation Example 5) solution 5ml in ice, and dissolved by the NaBH 4 by about 20mg added 10 times. Two drops of aqueous ammonia were added to this solution to adjust the pH to 10 or more, and then a reduction reaction was performed at room temperature for 3 hours. After the reaction, the remaining NaBH 4 was decomposed by adding Amberlite MB-4 (organo) until hydrogen generation and the color change of the resin disappeared. The liquid layer was collected while washing the resin with water and dried using a rotary evaporator. 1 ml of methanol was added to this and dissolved, and again dried. This operation was repeated 5 times. Finally, the product dissolved in purified water was used as a reduced product solution.
　得られた還元物のＤＥをソモギー変法（澱粉糖関連工業分析法（株式会社食品化学新聞社）（平成３年１１月１日発行）１１～１３頁参照）に従って測定したところ、ＤＥは０であった。還元前のＤＥは１０．２であり、還元反応により還元末端が完全に還元されたことが確認できた。また、還元反応の前後で重合度組成にも変化がなく、グルコシド結合の切断などは認められなかった（データ省略）。 The DE of the obtained reduced product was measured according to a modified somogy method (starch sugar related industrial analysis method (Food Chemical Newspaper Co., Ltd.) (issued on November 1, 1991), pages 11 to 13). Met. The DE before the reduction was 10.2, and it was confirmed that the reducing end was completely reduced by the reduction reaction. In addition, the composition of the degree of polymerization did not change before and after the reduction reaction, and no glucoside bond breakage was observed (data not shown).
配合例１：緑茶飲料
　表４に示す配合（全体を１００とした場合の質量比率）にて緑茶飲料を製造した。分岐メガロ糖シラップは製造例５のシラップを用いた。セルデックスＳＬ－２０、セルデックスＢ－１００、フジオリゴＧ６７は、日本食品化工社製のものを用いた（以下、同様）。セルデックスＳＬ－２０とはシクロデキストリンを２０％含む液状または粉末状の製品であり、セルデックスＢ－１００は結晶β―シクロデキストリンである。フジオリゴＧ６７は、マルトヘキサオースおよびマルトヘプタオースを主成分とするシラップである。なお、シラップ状原料の添加比率は、固形分の質量に基づいて計算した（以下、同様）。
Figure JPOXMLDOC01-appb-T000004
 Formulation Example 1: Green tea beverage A green tea beverage was produced according to the formulation shown in Table 4 (mass ratio based on 100 as a whole). As the branched megalosaccharide syrup, the syrup of Production Example 5 was used. Celldex SL-20, Celldex B-100, and Fujioligo G67 were manufactured by Nippon Shokuhin Kako Co., Ltd. (hereinafter the same). Celdex SL-20 is a liquid or powder product containing 20% cyclodextrin, and Celdex B-100 is crystalline β-cyclodextrin. Fuji oligo G67 is syrup mainly composed of maltohexaose and maltoheptaose. In addition, the addition ratio of the syrup-like raw material was calculated based on the mass of the solid content (hereinafter the same).
Figure JPOXMLDOC01-appb-T000004
　配合した分岐メガロ糖シラップとフジオリゴＧ６７の糖組成と甘味度は表５の通りであった。糖組成の分析は、ＨＰＬＣを用いて分析した。分析条件は、試験例２と同様とした。甘味度の測定は、試験例５に記載の方法で行った。
Figure JPOXMLDOC01-appb-T000005
 Table 5 shows the sugar composition and sweetness of the blended megalosaccharide syrup and Fujioligo G67. The sugar composition was analyzed using HPLC. The analysis conditions were the same as in Test Example 2. The sweetness level was measured by the method described in Test Example 5.
Figure JPOXMLDOC01-appb-T000005
　１０人のパネラーにて、作製した緑茶飲料の官能評価を行い、苦味・渋味および味のバランスについて評価を行った。評価結果を表６に示す。
Figure JPOXMLDOC01-appb-T000006
 Sensory evaluation of the prepared green tea beverage was performed by 10 panelists, and the balance of bitterness / astringency and taste was evaluated. The evaluation results are shown in Table 6.
Figure JPOXMLDOC01-appb-T000006
　表中の苦味・渋味低減効果については、非常に効果あり（◎）、効果あり（○）、ほとんど効果なし（△）、効果なし（×）の評価結果で示し、甘味については甘味が強い（◎）、すこし甘い（○）、甘くない（×）、旨味については旨味あり（◎）、旨味が少しあり（○）、旨味が少ない（△）、旨味が弱い（×）、総合評価については、非常に良い（◎）、良い（○）、あまり良くない（△）、悪い（×）の評価結果で示した。 The bitterness and astringency reduction effects in the table are shown as evaluation results of very effective (◎), effective (○), almost no effect (△), no effect (×), and sweetness is strong. (◎), slightly sweet (○), not sweet (×), umami is umami (◎), umami is a little (○), umami is low (△), umami is weak (x), overall evaluation The evaluation results were very good (良 い), good (良 い), not very good (△), and bad (×).
　表６に示すように、従来使用されているシクロデキストリンやシクロデキストリン混合物を使用した場合、公知のように苦味・渋味の低減が認められたが、旨味も弱くなったために全体の味のバランスが悪くなった。フジオリゴＧ６７を使用することで、その甘味によりポリフェノールの苦味・渋味が低減されが、甘味は緑茶にはそぐわないために総合評価は悪くなった。 As shown in Table 6, when a conventionally used cyclodextrin or cyclodextrin mixture was used, a decrease in bitterness and astringency was recognized as is well known, but since the umami became weak, the balance of the overall taste Became worse. By using Fujioligo G67, the sweet taste reduced the bitterness and astringency of polyphenols, but the sweetness was not suitable for green tea, so the overall evaluation deteriorated.
　一方、本発明による分岐メガロ糖シラップを使用するとポリフェノールの苦味・渋味を抑制することが可能であった。さらに旨味の付与も可能であり、甘味も呈さないので全体の味のバランスを損なうこともなく高い評価を得られた。 On the other hand, when the branched megalosaccharide syrup according to the present invention was used, it was possible to suppress the bitterness and astringency of polyphenols. Furthermore, umami can be imparted, and since it does not exhibit sweetness, a high evaluation was obtained without impairing the balance of the overall taste.
配合例２：高濃度ポリフェノール類含有緑茶飲料
　表７に示す配合（全体を１００とした場合の質量比率）にて高濃度ポリフェノール類含有緑茶飲料を製造した。分岐メガロ糖シラップは製造例５のシラップを用いた。ポリフェノンＣＨは三井農林社製のものを用いた（以下、同様）。
Figure JPOXMLDOC01-appb-T000007
 Formulation Example 2: High-concentration polyphenols-containing green tea beverage A high-concentration polyphenol-containing green tea beverage was produced according to the formulation shown in Table 7 (mass ratio when the whole was 100). As the branched megalosaccharide syrup, the syrup of Production Example 5 was used. Polyphenon CH manufactured by Mitsui Norin Co., Ltd. was used (hereinafter the same).
Figure JPOXMLDOC01-appb-T000007
　１０人のパネラーにて、作製した茶飲料の官能評価を行い、苦味・渋味および味のバランスについて評価を行った。評価結果を表８に示す。
Figure JPOXMLDOC01-appb-T000008
 Ten panelists performed sensory evaluation of the produced tea beverage and evaluated the balance of bitterness / astringency and taste. Table 8 shows the evaluation results.
Figure JPOXMLDOC01-appb-T000008
　表中の苦味・渋味低減効果については、非常に効果あり（◎）、効果あり（○）、ほとんど効果なし（△）、効果なし（×）の評価結果で示し、甘味については甘味が強い（◎）、すこし甘い（○）、甘くない（×）、旨味については旨味あり（◎）、旨味が少しあり（○）、旨味が少ない（△）、旨味が弱い（×）、総合評価については、非常に良い（◎）、良い（○）、あまり良くない（△）、悪い（×）の評価結果で示した。 The bitterness and astringency reduction effects in the table are shown as evaluation results of very effective (◎), effective (○), almost no effect (△), no effect (×), and sweetness is strong. (◎), slightly sweet (○), not sweet (×), umami is umami (◎), umami is a little (○), umami is low (△), umami is weak (x), overall evaluation The evaluation results were very good (良 い), good (良 い), not very good (△), and bad (×).
　表８に示すように、先の試験と同様、フジオリゴＧ６７やセルデックスＳＬ－２０やセルデックスＢ－１００を使用することで、ポリフェノールの苦味・渋味が低減された。しかしながら、フジオリゴＧ６７は甘味を呈し、セルデックスＳＬ－２０やセルデックスＢ－１００は旨味も低減してしまい全体の味のバランスが悪くなった。 As shown in Table 8, as in the previous test, the use of Fujioligo G67, Celldex SL-20, or Celldex B-100 reduced the bitterness and astringency of polyphenols. However, Fujioligo G67 exhibited sweetness, and Celdex SL-20 and Celdex B-100 also reduced the umami taste, resulting in a poor overall taste balance.
　一方、本発明による分岐メガロ糖シラップを使用するとポリフェノール類の苦味・渋味が低減可能であり、かつ旨味の付与も可能であった。さらにこれらの効果は茶の味のバランスを崩すことなく得られた。 On the other hand, when the branched megalosaccharide syrup according to the present invention is used, the bitterness and astringency of polyphenols can be reduced, and umami can be imparted. Furthermore, these effects were obtained without breaking the balance of tea taste.
配合例３：紅茶飲料
　表９に示す配合（全体を１００とした場合の質量比率）にて紅茶飲料を製造した。紅茶は、紅茶葉２ｇに対して２００ｍｌの湯（９５℃）を加え２分間抽出を行い、抽出後、吸引ろ過を行い得られた清澄な紅茶抽出液を用いた。分岐メガロ糖シラップは製造例５のシラップを用いた。
Figure JPOXMLDOC01-appb-T000009
 Formulation Example 3: Black Tea Beverage A black tea beverage was produced according to the formulation shown in Table 9 (mass ratio based on 100 as a whole). The black tea used was a clear black tea extract obtained by adding 200 ml of hot water (95 ° C.) to 2 g of black tea leaves and performing extraction for 2 minutes, followed by suction filtration after extraction. As the branched megalosaccharide syrup, the syrup of Production Example 5 was used.
Figure JPOXMLDOC01-appb-T000009
　１０人のパネラーにて、作製した紅茶飲料の官能評価を行い、苦味・渋味および味のバランスについて評価を行った。評価結果を表１０に示す。
Figure JPOXMLDOC01-appb-T000010
 Sensory evaluation of the produced black tea beverage was performed by 10 panelists, and the balance of bitterness / astringency and taste was evaluated. Table 10 shows the evaluation results.
Figure JPOXMLDOC01-appb-T000010
　表中の苦味・渋味低減効果については、非常に効果あり（◎）、効果あり（○）、ほとんど効果なし（△）、効果なし（×）の評価結果で示し、味のバランスについては、非常に良い（◎）、良い（○）、あまり良くない（△）、悪い（×）の評価結果で示した。 The bitterness and astringency reduction effects in the table are shown as evaluation results of very effective (◎), effective (○), almost no effect (△), no effect (×). The evaluation results were very good (◎), good (○), not very good (△), and bad (×).
　表１０に示すように、フジオリゴＧ６７やセルデックスＳＬ－２０やセルデックスＢ－１００を使用することで、ポリフェノールの苦味・渋味が低減された。しかしながら、フジオリゴＧ６７は甘味が感じられるため、全体的な味のバランスが悪くなった。また、セルデックスＳＬ－２０やセルデックスＢ－１００は紅茶の風味や旨味も低減してしまい、同様に全体の味のバランスが悪くなった。 As shown in Table 10, the use of Fujioligo G67, Celdex SL-20 and Celdex B-100 reduced the bitterness and astringency of polyphenols. However, since Fuji Oligo G67 felt sweet, the overall taste balance deteriorated. In addition, Celdex SL-20 and Celdex B-100 also reduced the flavor and umami of black tea, and similarly the overall taste balance was poor.
　一方、本発明による分岐メガロ糖シラップを使用するとまろやかになることが明らかとなり、全体の味のバランスを損ねることなくポリフェノールの苦味・渋味を抑制することが可能であった。 On the other hand, it was clarified that the branched megalosaccharide syrup according to the present invention was mellow, and it was possible to suppress the bitterness and astringency of polyphenols without impairing the overall taste balance.
配合例４：緑茶ゼリー
　表１１に示す配合（全体を１００とした場合の質量比率）で緑茶ゼリーを常法にて製造した。分岐メガロ糖シラップは製造例５のシラップを用いた。
Figure JPOXMLDOC01-appb-T000011
 Formulation Example 4: Green tea jelly Green tea jelly was prepared in a conventional manner with the formulation shown in Table 11 (mass ratio based on 100 as a whole). As the branched megalosaccharide syrup, the syrup of Production Example 5 was used.
Figure JPOXMLDOC01-appb-T000011
　１０人のパネラーにて、作製した緑茶ゼリーの官能評価を行い、苦味・渋味および味のバランスについて評価を行った。評価結果を表１２に示す。
Figure JPOXMLDOC01-appb-T000012
 The sensory evaluation of the produced green tea jelly was performed by 10 panelists, and the balance of bitterness / astringency and taste was evaluated. The evaluation results are shown in Table 12.
Figure JPOXMLDOC01-appb-T000012
　表中の苦味・渋味低減効果については、非常に効果あり（◎）、効果あり（○）、ほとんど効果なし（△）、効果なし（×）の評価結果で示し、コクについてはコクがあり（○）、コクがない（×）、味のバランスについては、非常に良い（◎）、良い（○）、あまり良くない（△）、悪い（×）の評価結果で示した。 The bitterness and astringency reduction effects in the table are shown as evaluation results of very effective (◎), effective (○), almost no effect (△), no effect (×). About ((circle)), there is no richness (x), and the balance of the taste was shown by the evaluation result of very good ((double-circle)), good ((circle)), not so good ((triangle | delta)), and bad (*).
　表１２に示すように、フジオリゴＧ６７やセルデックスＳＬ－２０やセルデックスＢ－１００を使用することで、ポリフェノールの苦味・渋味が低減された。しかしながら、フジオリゴＧ６７の低減効果はメガロ糖に比べて弱いものであった。また、セルデックスＳＬ－２０やセルデックスＢ－１００はメガロ糖と同等以上に苦味・渋味を低減したものの、緑茶の風味が感じにくくなった。 As shown in Table 12, the bitterness and astringency of polyphenols were reduced by using Fujioligo G67, Celdex SL-20 and Celdex B-100. However, the reduction effect of Fujioligo G67 was weak compared to megalosaccharide. In addition, although Celdex SL-20 and Celdex B-100 reduced bitterness and astringency more than or equal to megalo sugar, the flavor of green tea became difficult to feel.
　一方、本発明による分岐メガロ糖シラップを使用するとポリフェノールの苦味・渋味を抑制することが可能であった。かつ緑茶の味がまろやかに感じられ、さらに全体の味のバランスを損ねることもなかった。 On the other hand, when the branched megalosaccharide syrup according to the present invention was used, it was possible to suppress the bitterness and astringency of polyphenols. The taste of green tea was mellow, and the balance of the overall taste was not compromised.
配合例５：緑茶飲料
　表１３に示す配合（全体を１００とした場合の質量比率）にて緑茶飲料を製造した。分岐メガロ糖シラップは製造例１７で製造した分岐メガロ糖シラップ分画品を用いた。
Figure JPOXMLDOC01-appb-T000013
 Formulation Example 5: Green tea beverage A green tea beverage was produced according to the formulation shown in Table 13 (mass ratio based on 100 as a whole). As the branched megalosaccharide syrup, the branched megalosaccharide syrup fraction produced in Production Example 17 was used.
Figure JPOXMLDOC01-appb-T000013
　配合した分岐メガロ糖シラップ分画品とフジオリゴＧ６７の糖組成の分析結果は以下の通りであった。糖組成の分析は、ＨＰＬＣを用いて分析した。分析条件は、試験例２と同様とした。
Figure JPOXMLDOC01-appb-T000014
 The analysis results of the sugar composition of the blended megalosaccharide syrup fraction and Fujioligo G67 were as follows. The sugar composition was analyzed using HPLC. The analysis conditions were the same as in Test Example 2.
Figure JPOXMLDOC01-appb-T000014
　２３人のパネラーにて、作製した緑茶飲料の官能評価を行い、苦味・渋味および味のバランスについて評価を行った。評価結果を表１５に示す。
Figure JPOXMLDOC01-appb-T000015
 The sensory evaluation of the prepared green tea beverage was performed by 23 panelists, and the balance of bitterness / astringency and taste was evaluated. The evaluation results are shown in Table 15.
Figure JPOXMLDOC01-appb-T000015
　表中の苦味・渋味低減効果については、非常に効果あり（◎）、効果あり（○）、ほとんど効果なし（△）、効果なし（×）の評価結果で示し、甘味については甘味が強い（◎）、すこし甘い（○）、甘くない（×）、旨味については旨味あり（◎）、旨味が少しあり（○）、旨味が少ない（△）、旨味がない（×）、味のバランスについては、非常に良い（◎）、良い（○）、あまり良くない（△）、悪い（×）の評価結果で示した。 The bitterness and astringency reduction effects in the table are shown as evaluation results of very effective (◎), effective (○), almost no effect (△), no effect (×), and sweetness is strong. (◎), slightly sweet (○), not sweet (×), umami is umami (◎), umami is slightly (○), umami is low (△), umami is not (×), balance of taste The evaluation results were very good (◎), good (○), not very good (△), and bad (×).
　表１５に示すように、従来使用されているシクロデキストリン混合物を使用した場合、公知のように苦味・渋味の低減が認められたが、旨味も弱くなったために全体の味のバランスが悪くなった。フジオリゴＧ６７にも苦味・渋味低減効果が見られたが、甘味を呈したため茶飲料としては好まれない結果となった。 As shown in Table 15, when a conventionally used cyclodextrin mixture was used, a decrease in bitterness and astringency was recognized as is well known, but since the umami was weakened, the overall taste balance deteriorated. It was. Fujioligo G67 also showed a bitter and astringent taste reducing effect, but because of its sweet taste, it was not preferred as a tea beverage.
　一方、本発明による分岐メガロ糖シラップ分画品を使用すると全体の味のバランスを悪くすること無くポリフェノールの苦味・渋味を抑制することが可能であった。特開２００６－２８０２５４号公報では分岐構造を有する３～４糖類を有効成分としカテキンの苦味・渋味を抑制する方法が開示されているが、３～４糖類をほとんど含まない本発明による分岐メガロ糖シラップ分画品を使用した場合においても、苦味・渋味低減効果は認められた。また、３～４糖類をほとんど含まない本発明による分岐メガロ糖シラップを使用することで高い旨味の付与効果が得られた。 On the other hand, when the branched megalosaccharide syrup fraction product according to the present invention was used, it was possible to suppress the bitterness and astringency of polyphenols without deteriorating the overall taste balance. Japanese Patent Application Laid-Open No. 2006-280254 discloses a method for suppressing the bitterness and astringency of catechin using a 3-4 saccharide having a branched structure as an active ingredient. Even when the sugar syrup fraction was used, the bitterness / astringency reduction effect was recognized. Further, the use of the branched megalosaccharide syrup according to the present invention, which contains almost no 3-4 saccharides, gave a high umami imparting effect.
配合例６：高濃度ポリフェノール類含有緑茶飲料
　表１６に示す配合（全体を１００とした場合の質量比率）にて高濃度ポリフェノール類含有緑茶飲料を製造した。分岐メガロ糖シラップは製造例１７で製造した分岐メガロ糖シラップ分画品を用いた。
Figure JPOXMLDOC01-appb-T000016
 Formulation Example 6: High concentration polyphenols-containing green tea beverage A high concentration polyphenols-containing green tea beverage was produced according to the formulation shown in Table 16 (mass ratio when the whole was 100). As the branched megalosaccharide syrup, the branched megalosaccharide syrup fraction produced in Production Example 17 was used.
Figure JPOXMLDOC01-appb-T000016
　９人のパネラーにて、作製した茶飲料の官能評価を行い、苦味・渋味および味のバランスについて評価を行った。評価結果を表１７に示す。
Figure JPOXMLDOC01-appb-T000017
 Nine panelists performed sensory evaluation of the produced tea beverages, and evaluated the balance of bitterness / astringency and taste. The evaluation results are shown in Table 17.
Figure JPOXMLDOC01-appb-T000017
　表中の苦味・渋味低減効果については、非常に効果あり（◎）、効果あり（○）、ほとんど効果なし（△）、効果なし（×）の評価結果で示し、味のバランスについては、非常に良い（◎）、良い（○）、あまり良くない（△）、悪い（×）の評価結果で示した。 The bitterness and astringency reduction effects in the table are shown as evaluation results of very effective (◎), effective (○), almost no effect (△), no effect (×). The evaluation results were very good (◎), good (○), not very good (△), and bad (×).
　表１７に示すように、先の試験と同様、フジオリゴＧ６７やセルデックスＳＬ－２０を使用することで、ポリフェノールの苦味・渋味が低減された。しかしながら、フジオリゴＧ６７は甘味を呈し、セルデックスＳＬ－２０は旨味も低減してしまい全体の味のバランスが悪くなった。 As shown in Table 17, the bitterness and astringency of polyphenols were reduced by using Fujioligo G67 and Celdex SL-20, as in the previous test. However, Fujioligo G67 exhibited sweetness, and Celdex SL-20 also reduced umami, resulting in a poor overall taste balance.
　一方、本発明による分岐メガロ糖シラップ分画品を使用すると全体の味のバランスを損ねることなくポリフェノールの苦味・渋味を抑制することが可能であった。さらに配合例５と同様に、３～４糖類をほとんど含まない本発明による分岐メガロ糖シラップを使用した場合においても、苦味・渋味低減効果は認められた。 On the other hand, when the branched megalosaccharide syrup fraction product according to the present invention was used, it was possible to suppress the bitterness and astringency of polyphenols without impairing the overall taste balance. Further, as in Formulation Example 5, even when the branched megalosaccharide syrup according to the present invention containing almost no 3-4 saccharides was used, the bitterness / astringency reduction effect was recognized.
配合例７：紅茶飲料
　表１８に示す配合（全体を１００とした場合の質量比率）にて紅茶飲料を製造した。紅茶は、紅茶葉２ｇに対して２００ｍｌの湯（９５℃）を加え２分間抽出を行い、抽出後、吸引ろ過を行い得られた清澄な紅茶抽出液を用いた。分岐メガロ糖シラップは製造例１７で製造した分岐メガロ糖シラップ分画品を用いた。
Figure JPOXMLDOC01-appb-T000018
 Formulation Example 7: Black tea beverage A black tea beverage was produced according to the formulation shown in Table 18 (mass ratio based on 100 as a whole). The black tea used was a clear black tea extract obtained by adding 200 ml of hot water (95 ° C.) to 2 g of black tea leaves and performing extraction for 2 minutes, followed by suction filtration after extraction. As the branched megalosaccharide syrup, the branched megalosaccharide syrup fraction produced in Production Example 17 was used.
Figure JPOXMLDOC01-appb-T000018
　１０人のパネラーにて、作製した紅茶飲料の官能評価を行い、苦味・渋味および味のバランスについて評価を行った。評価結果を表１９に示す。
Figure JPOXMLDOC01-appb-T000019
 Sensory evaluation of the produced black tea beverage was performed by 10 panelists, and the balance of bitterness / astringency and taste was evaluated. The evaluation results are shown in Table 19.
Figure JPOXMLDOC01-appb-T000019
　表中の苦味・渋味低減効果については、非常に効果あり（◎）、効果あり（○）、ほとんど効果なし（△）、効果なし（×）の評価結果で示し、味のバランスについては、非常に良い（◎）、良い（○）、あまり良くない（△）、悪い（×）の評価結果で示した。 The bitterness and astringency reduction effects in the table are shown as evaluation results of very effective (◎), effective (○), almost no effect (△), no effect (×). The evaluation results were very good (◎), good (○), not very good (△), and bad (×).
　表１９に示すように、フジオリゴＧ６７やセルデックスＳＬ－２０やセルデックスＢ－１００を使用することで、ポリフェノールの苦味・渋味が低減された。しかしながら、フジオリゴＧ６７は甘味が感じられるとの評価を受け、結果として全体的な味のバランスが悪くなった。また、セルデックスＳＬ－２０やセルデックスＢ－１００は紅茶の風味や旨味も低減してしまい、同様に全体の味のバランスが悪くなった。 As shown in Table 19, the bitterness and astringency of polyphenols were reduced by using Fujioligo G67, Celdex SL-20, or Celdex B-100. However, Fuji Oligo G67 was evaluated as having a sweet taste, and as a result, the overall taste balance deteriorated. In addition, Celdex SL-20 and Celdex B-100 also reduced the flavor and umami of black tea, and similarly the overall taste balance was poor.
　一方、本発明による分岐メガロ糖シラップ分画品を使用すると全体の味のバランスを損ねることなくポリフェノールの苦味・渋味を抑制することが可能であった。また、評価コメントにあるように味をまろやかにする効果が認められた。 On the other hand, when the branched megalosaccharide syrup fraction product according to the present invention was used, it was possible to suppress the bitterness and astringency of polyphenols without impairing the overall taste balance. In addition, as shown in the evaluation comments, the effect of mellow taste was recognized.
配合例８：緑茶ゼリー
　表２０に示す配合（全体を１００とした場合の質量比率）で緑茶ゼリーを常法にて製造した。分岐メガロ糖シラップは製造例１７で製造した分岐メガロ糖シラップ分画品を用いた。
Figure JPOXMLDOC01-appb-T000020
 Formulation Example 8: Green tea jelly Green tea jelly was prepared in a conventional manner with the formulation shown in Table 20 (mass ratio based on 100 as a whole). As the branched megalosaccharide syrup, the branched megalosaccharide syrup fraction produced in Production Example 17 was used.
Figure JPOXMLDOC01-appb-T000020
　１０人のパネラーにて、作製した緑茶ゼリーの官能評価を行い、苦味・渋味および味のバランスについて評価を行った。評価結果を表２１に示す。
Figure JPOXMLDOC01-appb-T000021
 The sensory evaluation of the produced green tea jelly was performed by 10 panelists, and the balance of bitterness / astringency and taste was evaluated. The evaluation results are shown in Table 21.
Figure JPOXMLDOC01-appb-T000021
　表中の苦味・渋味低減効果については、非常に効果あり（◎）、効果あり（○）、ほとんど効果なし（△）、効果なし（×）の評価結果で示し、コクの付与は効果あり（○）、効果なし（×）、味のバランスについては、非常に良い（◎）、良い（○）、あまり良くない（△）、悪い（×）の評価結果で示した。 The bitterness and astringency reduction effects in the table are shown as evaluation results of very effective (◎), effective (○), almost no effect (△), no effect (×), and richness is effective The evaluation results of (◯), no effect (×), and balance of taste were shown as evaluation results of very good (◎), good (○), not very good (Δ), and bad (×).
　表２１に示すように、フジオリゴＧ６７やセルデックスＳＬ－２０やセルデックスＢ－１００を使用することで、ポリフェノールの苦味・渋味が低減された。しかしながら、フジオリゴＧ６７の低減効果は分岐糖類に比べて弱いものであった。また、セルデックスＳＬ－２０やセルデックスＢ－１００は分岐メガロ糖シラップと同等以上に苦味・渋味を低減したものの、緑茶の風味が感じにくくなった。 As shown in Table 21, the bitterness and astringency of polyphenols were reduced by using Fujioligo G67, Celdex SL-20, or Celdex B-100. However, the reduction effect of Fujioligo G67 was weaker than that of branched sugars. In addition, Celdex SL-20 and Celdex B-100 reduced bitterness and astringency as much as or more than the branched megalosaccharide syrup, but the green tea flavor became difficult to feel.
　一方、本発明による分岐メガロ糖シラップ分画品を使用するとポリフェノール類の苦味・渋味を抑制することが可能であった。かつコクの付与も可能であり、本発明による分岐メガロ糖シラップ分画品の使用は全体の味のバランスを損ねることもなかった。 On the other hand, when the branched megalosaccharide syrup fraction according to the present invention was used, it was possible to suppress the bitterness and astringency of polyphenols. In addition, richness can be imparted, and the use of the branched megalosaccharide syrup fraction according to the present invention did not impair the overall taste balance.
配合例９：果汁飲料
　市販低果汁飲料に対し、製造例１で調製した分岐メガロ糖シラップを１％（ｗ／ｗ）（固形分換算）配合した果汁飲料を調製した。その結果、未添加区と比較して分岐メガロ糖添加区ではコク味が増し、果汁飲料に深みを与えることが可能であった。分岐メガロ糖は低果汁飲料などにおいても、コク味増強剤として有利に使用することができる。 Formulation Example 9: Fruit juice beverage A fruit juice beverage was prepared by blending 1% (w / w) (in terms of solid content) of the branched megalosaccharide syrup prepared in Production Example 1 with respect to a commercially available low fruit juice beverage. As a result, compared to the non-added group, the richness increased in the branched megalosaccharide added group, and it was possible to give depth to the fruit juice drink. Branched megalosaccharide can be advantageously used as a rich taste enhancer even in a low fruit juice beverage.
配合例１０：高甘味度甘味料含有飲料
　アセスルファムＫ０．０５％（ｗ／ｗ）もしくはアスパルテーム０．０３％（ｗ／ｗ）含有飲料に対し、製造例１で調製した分岐メガロ糖シラップを１％（ｗ／ｗ）（固形分換算）配合した飲料を調製した。分岐メガロ糖の後味のほのかな甘さが効果的に働き、高甘味度甘味料の後味の苦味を改善し、飲み心地の良い飲料とすることが可能であった。このように本発明による分岐メガロ糖は高甘味度甘味料の味質改善剤として有利に使用することができる。 Formulation Example 10: High-sweetness sweetener-containing beverage Acesulfame K 0.05% (w / w) or aspartame 0.03% (w / w) -containing beverage 1% of the branched megalosaccharide syrup prepared in Production Example 1 A blended beverage (w / w) (solid content conversion) was prepared. The subtle sweetness of the aftertaste of branched megalo sugar worked effectively, and the bitterness of the aftertaste of high-intensity sweeteners was improved, and it was possible to make a drink that was comfortable to drink. Thus, the branched megalosaccharide according to the present invention can be advantageously used as a taste improving agent for high-intensity sweeteners.
配合例１１：冷凍卵焼き
　全卵１００質量部に対し、製造例１にて調製した分岐メガロ糖シラップ７質量部（固形分換算）、加工デンプン１．５質量部、出し汁３０質量部を混合し、常法にて卵焼きを作製した。急速凍結機で凍結させ、２週間後に室温にて解凍した。本品は耐老化性に優れる分岐メガロ糖を含有していることから、冷凍解凍後であるにも関わらず、みずみずしさを保っていた。すなわち、本発明による分岐メガロ糖は、冷凍解凍後のみずみずしさを維持できる効果を有していることが確認された。 Formulation Example 11: With respect to 100 parts by mass of the frozen fried egg whole eggs, 7 parts by mass of the branched megalosugar syrup prepared in Production Example 1 (in terms of solid content), 1.5 parts by mass of the modified starch, and 30 parts by mass of the soup stock are mixed. Fried eggs were prepared by a conventional method. It was frozen with a quick freezer and thawed at room temperature after 2 weeks. Since this product contains a branched megalosaccharide having excellent aging resistance, it was kept fresh even after freezing and thawing. That is, it was confirmed that the branched megalosaccharide according to the present invention has an effect of maintaining freshness after freezing and thawing.
配合例１２：パン
　強力粉１００質量部に対し、各対粉イースト２．２質量部、イーストフード０．１質量部、砂糖２質量部、製造例１にて調製した分岐メガロ糖シラップ３質量部（固形分換算）、食塩２質量部、脱脂粉乳２質量部、油脂６質量部、水６７質量部を使用し、中種法にて食パンを作製した。本品は耐老化性に優れる分岐メガロ糖を含有していることから、ふんわりとした食感の食パンが得られた。すなわち、本発明による分岐メガロ糖は、食品にふんわりとした食感を付与できる効果を有していることが確認された。 Formulation Example 12: 2.2 parts by mass of each anti-powder yeast, 0.1 parts by mass of yeast food, 2 parts by mass of sugar, 3 parts by mass of the branched megalosaccharide syrup prepared in Production Example 1 (100 parts by mass of bread crumbs ) Solid content conversion), 2 parts by mass of sodium chloride, 2 parts by mass of skim milk powder, 6 parts by mass of fats and oils, and 67 parts by mass of water were used to prepare bread using a medium seed method. Since this product contains a branched megalosaccharide with excellent aging resistance, a bread with a soft texture was obtained. That is, it was confirmed that the branched megalosaccharide according to the present invention has an effect of giving a soft texture to food.
配合例１３：ソフトクッキー
　油脂対粉１７質量部に、製造例１にて調製した分岐メガロ糖シラップ（固形分換算）対粉５０質量部、砂糖対粉６０質量部、全卵対粉５０質量部を順次添加し混合した。バニラオイルを対粉０．２質量部加えた後、脱脂粉乳対粉４０質量部および予め篩っておいた小麦粉１００およびベーキングパウダー対粉０．９質量部を混合した。上火１９５℃、下火１７０℃で１３分焼成した。本品は耐老化性に優れる分岐メガロ糖を含有していることから、長期間しっとりした食感を保持可能であった。すなわち、本発明による分岐メガロ糖は、食品にしっとりした食感を付与できる効果を有していることが確認された。 Formulation Example 13: 17 parts by mass of soft cookie oil / fat, 50 parts by mass of branched megalosugar syrup (solid equivalent) prepared in Production Example 1 vs. powder, 60 parts by mass of sugar vs. powder, 50 parts by mass of whole egg Were sequentially added and mixed. After adding 0.2 parts by weight of vanilla oil to powder, 40 parts by weight of skim milk powder and powder, 100 sieved wheat flour and 0.9 parts by weight of baking powder were mixed. Firing was carried out for 13 minutes at 195 ° C. on the top and 170 ° C. on the bottom. Since this product contains a branched megalosaccharide with excellent aging resistance, it can retain a moist texture for a long period of time. That is, it was confirmed that the branched megalosaccharide according to the present invention has an effect of imparting a moist texture to food.
配合例１４：氷の均一性向上・促進効果
　製造例５にて調製した分岐メガロ糖シラップを４％（ｗ／ｗ）（固形分換算）、砂糖を５％（ｗ／ｗ）、色素を０．４％（ｗ／ｗ）（サンレッドＮｏ．５Ｆ（三栄源エフ・エフ・アイ社製））それぞれ添加して混合水溶液を調製した。これを－３０℃の冷凍庫にて一晩凍結させた。室温にて融解時に経時的にサンプリングし、高速液体クロマトグラフィー（ＨＰＬＣ）にて分岐メガロ糖および砂糖の溶出状態を確認した。ＨＰＬＣの条件としてはカラムにＵｌｔｒｏｎ　ＰＳ－８０Ｎ・Ｌ（Φ８．０ｍｍ×５００ｍｍ、信和化工社製）を用い、移動相を超純水とし、カラム温度５０℃、流速０．９ｍｌ／分とした。分析では３～５倍希釈液を２μｌ程度供した。その結果、本発明による分岐メガロ糖を使用することで無添加区と比較して砂糖の溶出が緩やかになることが明らかとなった。 Formulation Example 14: Ice uniformity improvement / promotion effect 4% (w / w) of branched megalosaccharide syrup prepared in Production Example 5 (in terms of solid content), 5% of sugar (w / w), and 0 of pigment .4% (w / w) (Sun Red No. 5F (manufactured by San-Ei Gen FFI Co., Ltd.)) was added to prepare a mixed aqueous solution. This was frozen overnight in a −30 ° C. freezer. Sampling was carried out over time at the time of melting at room temperature, and the elution state of branched megalosaccharide and sugar was confirmed by high performance liquid chromatography (HPLC). As HPLC conditions, Ultra PS-80N · L (Φ8.0 mm × 500 mm, manufactured by Shinwa Kako Co., Ltd.) was used for the column, the mobile phase was ultrapure water, the column temperature was 50 ° C., and the flow rate was 0.9 ml / min. In the analysis, about 2 μl of a 3 to 5-fold diluted solution was used. As a result, it has been clarified that the use of the branched megalosaccharide according to the present invention makes the dissolution of sugar gentler than in the non-added group.
配合例１５：酢飲料
　りんご酢８％（ｗ／ｗ）、果糖ぶどう糖液糖（日本食品化工社製）８％（ｗ／ｗ）（固形分換算）および製造例１で調製した分岐メガロ糖シラップ１％（ｗ／ｗ）（固形分換算）を配合した酢飲料を評価した。本品は酸味低減効果に優れる分岐メガロ糖を含有していることから、酸味が和らぎ、飲み心地を向上させることが可能であった。すなわち、本発明による分岐メガロ糖は酸味の低減効果を有していることが確認された。 Formulation Example 15: Vinegar Beverage Apple Vinegar 8% (w / w), Fructose Glucose Liquid Sugar (manufactured by Nippon Shokuhin Kako Co., Ltd.) 8% (w / w) (in terms of solid content) and Branched Megalosaccharide Syrup The vinegar drink which mix | blended 1% (w / w) (solid content conversion) was evaluated. Since this product contains a branched megalosaccharide that has an excellent acidity-reducing effect, the acidity is softened and the drinking comfort can be improved. That is, it was confirmed that the branched megalosaccharide according to the present invention has a sour taste reducing effect.
配合例１６：無菌米飯
　米１００質量部に水１５４．５質量部、グルコノデルタラクトン０．７５質量部および製造例１で調製した分岐メガロ糖シラップ２質量部（固形分換算）を混合し、炊飯した。酸臭抑制効果に優れる分岐メガロ糖を含有していることから、炊飯後もしくは再加熱時のグルコノデルタラクトン由来の酸臭を低減した。すなわち、本発明による分岐メガロ糖は、酸臭を低減する効果を有しており、保存剤の酸臭低減剤として使用可能である。 Formulation Example 16: Mixing 154.5 parts by mass of water, 0.75 parts by mass of gluconodeltalactone and 2 parts by mass of the branched megalosaccharide syrup prepared in Production Example 1 (in terms of solid content) with 100 parts by mass of aseptic cooked rice, I cooked rice. Since it contains a branched megalosaccharide that has an excellent acid odor suppressing effect, the acid odor derived from gluconodeltalactone after cooking or during reheating was reduced. That is, the branched megalosaccharide according to the present invention has an effect of reducing the acid odor, and can be used as an acid odor reducing agent for a preservative.
配合例１７：アルコール飲料
　ニコライウォッカ（キリンシーグラム社）を用いてアルコール濃度６％（ｖ／ｖ）となるように、果糖ぶどう糖液糖４％（ｗ／ｖ）（固形分換算）および製造例５で調製した分岐メガロ糖シラップ１％（ｗ／ｖ）（固形分換算）を配合したアルコール飲料を評価した。本品はアルコール感緩和に優れる分岐メガロ糖を含有していることから、アルコールの嫌味が低減され飲み心地が向上した。すなわち、本発明による分岐メガロ糖は、アルコール飲料の味質改善剤として有利に使用することができる。 Formulation Example 17: Fructose-glucose liquid sugar 4% (w / v) (in terms of solid content) and Production Example 5 so that the alcohol concentration is 6% (v / v) using Nikolai vodka (Kirin Seagram Co., Ltd.) The alcoholic beverage blended with 1% (w / v) branched megalosaccharide syrup (converted to solid content) prepared in the above was evaluated. Since this product contains branched megalosaccharide, which is excellent in reducing alcohol feeling, the taste of alcohol is reduced and the drinking comfort is improved. That is, the branched megalosaccharide according to the present invention can be advantageously used as a taste improving agent for alcoholic beverages.
配合例１８：乳飲料
　牛乳３０％（ｗ／ｗ）に果糖ぶどう糖液糖４％（ｗ／ｗ）（固形分換算）および製造例５で調製した分岐メガロ糖シラップ１％（ｗ／ｗ）（固形分換算）を混合し、乳飲料を調製した。本品は無添加区と比較して乳味感が向上しており、コクのある飲料となった。すなわち、本発明による分岐メガロ糖は、乳味感の向上を目的として、乳を含有する飲食品に使用することができる。 Formulation Example 18: Milk Beverage Milk 30% (w / w), Fructose Glucose Liquid Sugar 4% (w / w) (Solid Content) and Branched Megalosaccharide Syrup 1% (w / w) Prepared in Production Example 5 ( A solid beverage) was mixed to prepare a milk beverage. Compared to the additive-free zone, this product has improved milky taste and has a rich beverage. That is, the branched megalosaccharide by this invention can be used for the food-drinks containing milk for the purpose of the improvement of a milky taste.
配合例１９：ゼリー
　デキストリン（ＴＫ－１６（松谷化学工業製））２０％（ｗ／ｗ）、果糖ぶどう糖液糖８％（固形分換算）（日本食品化工製）、クールアガー（新田ゼラチン製）２％（ｗ／ｗ）、クエン酸三Ｎａ　０．１％（ｗ／ｗ）、クエン酸０．１７％（ｗ／ｗ）および製造例５で調製した分岐メガロ糖シラップ５％（ｗ／ｗ）（固形分換算）を使用し、ゼリーを調製した。ＴＫ－１６とはＤＥ１６～１９程度の粉末状デキストリンである。本品はデキストリンの粉臭低減効果に優れる分岐メガロ糖を含有していることから、本試験区のように多量のデキストリンを使用したゼリーにおいてもデキストリン臭が緩和された。すなわち、本発明による分岐メガロ糖は飲食品にデキストリンを多量に含有させた場合でも有利に使用することができる。 Formulation Example 19: Jelly dextrin (TK-16 (manufactured by Matsutani Chemical Industry)) 20% (w / w), fructose-glucose liquid sugar 8% (in terms of solid content) (manufactured by Nippon Shokuhin Kako), cool agar (manufactured by Nitta Gelatin) 2% (w / w), tri-Na tricitrate 0.1% (w / w), citric acid 0.17% (w / w) and 5% of branched megalosaccharide syrup prepared in Production Example 5 (w / w) ) (In terms of solid content) was used to prepare a jelly. TK-16 is a powdery dextrin having a DE of 16-19. Since this product contains a branched megalosaccharide that is excellent in reducing the powdery odor of dextrin, the dextrin odor was alleviated even in jelly using a large amount of dextrin as in this test section. That is, the branched megalosaccharide according to the present invention can be advantageously used even when the food or drink contains a large amount of dextrin.
配合例２０：照り焼きのタレ
　表２２に示す配合（水を２００とした場合の質量比率）にて照り焼きのタレを調製した。分岐メガロ糖シラップは製造例１のシラップを用いた。フジオリゴＧ６７は、マルトヘキサオースおよびマルトヘプタオースを主成分とするシラップである。鶏のモモ肉を使用し、肉の重量の３０％（ｗ／ｗ）となる量のタレに一晩浸漬した。この鶏肉を中心温度７０℃に達するまで蒸気加熱し、タレに再度どぶづけした後、１８０℃のホットプレートにて片面１．５分ずつ焼成した。
Figure JPOXMLDOC01-appb-T000022
 Blending Example 20: Teriyaki Sauce A Teriyaki sauce was prepared according to the blending shown in Table 22 (mass ratio when water was 200). As the branched megalosaccharide syrup, the syrup of Production Example 1 was used. Fuji oligo G67 is syrup mainly composed of maltohexaose and maltoheptaose. Chicken thighs were used and soaked overnight in a sauce that was 30% (w / w) of the weight of the meat. The chicken was steam-heated until it reached a central temperature of 70 ° C., and then poured again into the sauce, and then baked on a hot plate at 180 ° C. for 1.5 minutes on each side.
Figure JPOXMLDOC01-appb-T000022
　１０人のパネラーにて作製した照り焼きチキンの官能評価を行い、照り・つやの評価を行った。評価結果を表２３に示す。
Figure JPOXMLDOC01-appb-T000023
 The teriyaki chicken produced by 10 panelists was subjected to sensory evaluation, and shine and gloss were evaluated. The evaluation results are shown in Table 23.
Figure JPOXMLDOC01-appb-T000023
　表中の照り・つやについては、効果あり（○）、若干効果有り（△）、効果なし（×）の評価結果で示した。 Shine and gloss in the table are shown as evaluation results with an effect (O), with a slight effect (△), and with no effect (×).
　表２３に示すように照り・つや向上効果を有するとされているフジオリゴＧ６７を使用した場合、照り・つや向上が認められた。一方、本発明による分岐メガロ糖シラップを使用した場合には、フジオリゴＧ６７以上に照り・つやを向上させることが確認された。 As shown in Table 23, when Fuji Oligo G67, which is supposed to have an effect of improving shine and gloss, was used, improvement of shine and gloss was recognized. On the other hand, when the branched megalosaccharide syrup according to the present invention was used, it was confirmed that the shine and gloss were improved over those of Fuji Oligo G67.
配合例２１：乳タンパク質を含む飲料（１）
　表２４に示す配合（全体を４０とした場合の質量比率）にて乳タンパク質含有飲料を下記の手順にて製造した。すなわち、表２４に示す糖またはデキストリンと脱脂粉乳を溶解し、混合撹拌した。次いで、クエン酸、クエン酸三Ｎａ、アスコルビン酸を用いてｐＨを６．４に調整した。この水溶液を５０ｍｌ容のファルコンチューブに充填し、密閉して１２１℃・１．２ｋｇｆ／ｃｍ２で達温のレトルト殺菌（トミー精工社製ＨＩＧＨ－ＰＲＥＳＳＵＲＥ　ＳＴＥＡＭ　ＳＴＥＲＩＬＩＺＥＲ／ＢＳ－３２５を使用）を行った。
Figure JPOXMLDOC01-appb-T000024
 Formulation Example 21: Beverage containing milk protein (1)
A milk protein-containing beverage was produced according to the following procedure with the formulation shown in Table 24 (mass ratio when the whole was 40). That is, sugar or dextrin and skim milk powder shown in Table 24 were dissolved and mixed and stirred. Next, the pH was adjusted to 6.4 using citric acid, tri-Na citrate, and ascorbic acid. This aqueous solution was filled into a 50 ml Falcon tube, sealed and subjected to retort sterilization at 121 ° C. and 1.2 kgf / cm 2 (using HIGH-PRESSURE STEAM STERILIZER / BS-325 manufactured by Tommy Seiko Co., Ltd.). .
Figure JPOXMLDOC01-appb-T000024
　分岐メガロ糖シラップは製造例５で得られたシラップを用いた。デキストリンはパインデックス＃２（ＤＥ約１１　松谷化学工業製）、パインデックス＃１００（ＤＥ約２～５　松谷化学工業製）、フジスター５Ｖ（ＤＥ約５　日本食品化工製）、フジスター１２Ｖ（ＤＥ約１２　日本食品化工製）、パインデックス＃４（ＤＥ約１９　松谷化学工業製）、ＴＫ－１６（ＤＥ約１６～１９　松谷化学工業製）、クラスター　デキストリン（ＤＥ約２　日本食品化工製）を使用した。 As the branched megalosaccharide syrup, the syrup obtained in Production Example 5 was used. Dextrin is Paindex # 2 (DE approx. 11 manufactured by Matsutani Chemical Industry Co., Ltd.), Paindex # 100 (DE approx. 2-5 manufactured by Matsutani Chemical Industry Co., Ltd.), Fujistar 5V (DE approx. 5 manufactured by Nippon Shokuhin Kako), Fujister 12V (DE About 12 (made by Nippon Food Chemicals), Paindex # 4 (DE about 19; made by Matsutani Chemical Industry), TK-16 (DE about 16-19 made by Matsutani Chemical Industry), cluster dextrin (DE about 2 made by Nippon Food Chemicals) did.
　上記のようにして作製した飲料の乳タンパク質の凝集状態を目視にて確認し、安定性試験とした。その結果を表２５に示す。表中のタンパク質安定性については、安定（○）、少し安定（△）、不安定（×）の評価結果で示した。
Figure JPOXMLDOC01-appb-T000025
 The aggregation state of milk protein in the beverage produced as described above was visually confirmed, and a stability test was performed. The results are shown in Table 25. The protein stability in the table is indicated by the evaluation results of stability (◯), slightly stable (Δ), and unstable (×).
Figure JPOXMLDOC01-appb-T000025
　その結果、表２５に示すように、比較例３９、４１、４３～４５では水溶液が完全に透明とはならないものの、タンパク質が凝集し沈澱が見られた。また、比較例４０、４２では水溶液が完全に透明になり、タンパク質の凝集沈澱が見られた。一方、本発明による分岐メガロ糖シラップを使用した実施例１６ではタンパク質の凝集や沈殿物は見られず、水溶液が白濁したまま安定であった。 As a result, as shown in Table 25, in Comparative Examples 39, 41, and 43 to 45, although the aqueous solution was not completely transparent, the protein was aggregated and precipitation was observed. In Comparative Examples 40 and 42, the aqueous solution was completely transparent, and protein aggregation and precipitation were observed. On the other hand, in Example 16 using the branched megalosaccharide syrup according to the present invention, protein aggregation and precipitation were not observed, and the aqueous solution was stable while becoming cloudy.
配合例２２：乳タンパク質を含む飲料（２）
　表２６に示す配合（全体を２００とした場合の質量比率）にて乳タンパク質含有飲料を下記の手順にて製造した。すなわち、表２６に示す糖またはデキストリンと脱脂粉乳を溶解し、混合撹拌した。次いで、クエン酸、クエン酸三Ｎａ、アスコルビン酸を用いてｐＨを６．４に調整した。この水溶液を１００ｍｌ容のショット瓶に充填し、密閉して１２１℃・１．２ｋｇｆ／ｃｍ２で達温のレトルト殺菌（トミー精工社製ＨＩＧＨ－ＰＲＥＳＳＵＲＥ　ＳＴＥＡＭ　ＳＴＥＲＩＬＩＺＥＲ／ＢＳ－３２５を使用）を行った。
Figure JPOXMLDOC01-appb-T000026
 Formulation Example 22: Beverage containing milk protein (2)
A milk protein-containing beverage was produced according to the following procedure with the formulation shown in Table 26 (mass ratio when the whole was 200). That is, sugar or dextrin and skim milk powder shown in Table 26 were dissolved and mixed and stirred. Next, the pH was adjusted to 6.4 using citric acid, tri-Na citrate, and ascorbic acid. This aqueous solution was filled into a 100 ml shot bottle, sealed, and subjected to retort sterilization at a temperature of 121 ° C. and 1.2 kgf / cm 2 (using a HIGH-PRESS STEAM STERIZIZER / BS-325 manufactured by Tommy Seiko Co., Ltd.). .
Figure JPOXMLDOC01-appb-T000026
　分岐メガロ糖シラップは製造例５のシラップを使用し、デキストリンはパインデックス＃２（ＤＥ約１１　松谷化学工業製）を使用した。フジオリゴＧ６７は、マルトヘキサオースおよびマルトヘプタオースを主成分とするシラップである。 The branched megalosugar syrup used the syrup of Production Example 5, and the dextrin used Paindex # 2 (DE about 11 manufactured by Matsutani Chemical Industries). Fujioligo G67 is syrup mainly composed of maltohexaose and maltoheptaose.
　上記のようにして作製した飲料の乳タンパク質の凝集状態を目視にて確認し、安定性試験とした。また、使用した糖またはデキストリンの１０％（ｗ／ｗ）水溶液を調整し、Ｏｓｍｏｍｅｔｅｒ（Ｆｉｓｋｅ　Ｍａｒｋ３）にて浸透圧を測定した。その結果を表２７に示す。表中のタンパク質安定性については、安定（○）、不安定（×）の評価結果で示した。
Figure JPOXMLDOC01-appb-T000027
 The aggregation state of milk protein in the beverage produced as described above was visually confirmed, and a stability test was performed. In addition, a 10% (w / w) aqueous solution of sugar or dextrin used was prepared, and the osmotic pressure was measured with an Osmometer (Fiske Mark 3). The results are shown in Table 27. The protein stability in the table is indicated by the evaluation results of stability (◯) and instability (×).
Figure JPOXMLDOC01-appb-T000027
　その結果、表２７に示すように、比較例４６では乳タンパク質は安定して存在していたが、比較例４７では乳タンパク質の凝集が認められた。しかしながら、比較例４６に使用した糖質はデキストリンである比較例４７と比較して２倍以上の高浸透圧であった。一方、本発明によるメガロ糖シラップを使用すると乳タンパク質は安定して存在した。 As a result, as shown in Table 27, milk protein was stably present in Comparative Example 46, whereas in Comparative Example 47, aggregation of milk protein was observed. However, the saccharide used in Comparative Example 46 had a high osmotic pressure more than twice that of Comparative Example 47, which is a dextrin. On the other hand, milk protein was stably present when using the megalo sugar syrup according to the present invention.
　本発明による分岐メガロ糖は単糖類や二糖類、オリゴ糖と比較して低浸透圧に調整容易であり、胃腸への負担が少ない。また、デキストリンを使用したときのような乳タンパク質の不安定化も見られないため、浸透圧の調整が必要な経腸栄養剤や経口栄養剤に有利に使用することができる。 The branched megalosaccharide according to the present invention can be easily adjusted to a low osmotic pressure as compared with monosaccharides, disaccharides, and oligosaccharides, and has less burden on the gastrointestinal tract. Moreover, since the milk protein is not destabilized as when dextrin is used, it can be advantageously used for enteral and oral nutrients that require adjustment of osmotic pressure.

Claims (36)

  1. 　重合度１１～３５のグルカンであって、少なくとも非還元末端に分岐構造を有するグルカン、またはその還元物。 A glucan having a polymerization degree of 11 to 35 and having a branched structure at least at the non-reducing end, or a reduced product thereof.
  2. 　α－１，４－グルコシド結合により構成されたグルカンと、その非還元末端に結合した分岐構造とからなる、請求項１に記載のグルカンまたはその還元物。 The glucan or reduced product thereof according to claim 1, comprising a glucan composed of α-1,4-glucoside bonds and a branched structure bonded to a non-reducing end thereof.
  3. 　分岐構造が、α－１，４－グルコシド結合以外のグルコシド結合により非還元末端に結合した１個以上のグルコース残基により構成される、請求項２に記載のグルカンまたはその還元物。 The glucan or a reduced product thereof according to claim 2, wherein the branched structure is composed of one or more glucose residues bonded to the non-reducing end by a glucoside bond other than an α-1,4-glucoside bond.
  4. 　請求項１～３のいずれか一項に記載のグルカンまたはその還元物を固形分当たり１０～６０質量％含有する液糖または粉糖。 Liquid sugar or powdered sugar containing 10 to 60% by mass of the glucan or reduced product thereof according to any one of claims 1 to 3 per solid content.
  5. 　シクロデキストリン生成酵素と糖転移作用を有する酵素とを、デンプン原料に作用させる工程を含んでなる、請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖の製造法。 The glucan or reduced product thereof according to any one of claims 1 to 3, further comprising a step of allowing a cyclodextrin-forming enzyme and an enzyme having a glycosyltransferase action to act on a starch raw material. Production method of liquid sugar or powdered sugar.
  6. 　シクロデキストリン生成酵素と糖転移作用を有する酵素に加えて、枝切り酵素を更に作用させる、請求項５に記載の製造法。 6. The production method according to claim 5, wherein a debranching enzyme is further acted on in addition to the cyclodextrin-forming enzyme and the enzyme having a glycosyltransferase effect.
  7. 　シクロデキストリン生成酵素が、パエニバチルス エスピー (Paenibacillus sp.)、バチルス コアギュランス (Bacillus coagulans)、バチルス ステアロサーモフィルス (Bacillus stearothermophilus)、またはバチルス マゼランス (Bacillus macelans) 由来のものである、請求項５または６に記載の製造法。 Cyclodextrin synthase, Paenibacillus sp (Paenibacillus sp.), Bacillus coagulans (Bacillus coagulans), Bacillus stearothermophilus (Bacillus stearothermophilus), or Bacillus Mazeransu (Bacillus macelans) is derived from, to claim 5 or 6 The manufacturing method described.
  8. 　糖転移作用を有する酵素が、α－グルコシダーゼ、６－α－グルコシルトランスフェラーゼ、デキストリンデキストラナーゼ、または環状マルトシルマルトース生成酵素である、請求項５または６に記載の製造法。 The production method according to claim 5 or 6, wherein the enzyme having a sugar transfer action is α-glucosidase, 6-α-glucosyltransferase, dextrin dextranase, or cyclic maltosyl maltose producing enzyme.
  9. 　α-グルコシダーゼが、アスペルギルス ニガー (Aspergillus niger) またはアクレモニウム エスピー (Acremonium sp.) 由来のものである請求項８に記載の製造法。 The production method according to claim 8, wherein the α-glucosidase is derived from Aspergillus niger or Acremonium sp.
  10. 　枝切り酵素が、イソアミラーゼ、プルラナーゼ、およびこれらの組み合わせからなる群から選択される、請求項６に記載の製造法。 The production method according to claim 6, wherein the debranching enzyme is selected from the group consisting of isoamylase, pullulanase, and combinations thereof.
  11. 　枝切り酵素が、マイロイデス オドラータス (Myroides odoratus)由来イソアミラーゼ、シュードモナス アミロデラモサ (Pseudomonas amyloderamosa) 由来イソアミラーゼ、およびクレブシエラ プネウモニアエ（Klebsiella pneumoniae）由来プルラナーゼ、並びにこれらの組み合わせからなる群から選択される、請求項６に記載の製造法。 Claim 6 wherein the debranching enzyme is selected from the group consisting of an isoamylase derived from Myroides odoratus, an isoamylase derived from Pseudomonas amyloderamosa, a pullulanase derived from Klebsiella pneumoniae , and a combination thereof. The production method described in 1.
  12. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる、食品添加剤または製剤用添加剤。 A food additive or a pharmaceutical additive comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4.
  13. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる、食品用風味改善剤または製剤用マスキング剤。 A food flavor improving agent or a preparation masking agent comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4.
  14. 　風味改善が、苦味および／または渋味の低減、酸味の低減、エグ味および／または嫌味の低減、コク味の増強、不快臭の低減、またはアルコール感の低減である、請求項１３に記載の風味改善剤。 The flavor improvement is reduction of bitterness and / or astringency, reduction of acidity, reduction of taste and / or taste, enhancement of richness, reduction of unpleasant odor, or reduction of alcohol feeling. Flavor improver.
  15. 　風味改善が、苦味および／または渋味の低減である、請求項１４に記載の風味改善剤。 The flavor improving agent according to claim 14, wherein the flavor improvement is reduction of bitterness and / or astringency.
  16. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる、食感改善剤。 A texture-improving agent comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4.
  17. 　食感改善が、パサパサ感の抑制、しっとり感の付与、またはふんわりした食感の維持である、請求項１６に記載の食感改善剤。 The texture-improving agent according to claim 16, wherein the texture improvement is suppression of a crispy feeling, imparting a moist feeling, or maintaining a soft texture.
  18. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる、氷の均一的凍結剤。 A uniform ice freezing agent comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4.
  19. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる、食品の照りおよび／またはつや向上剤。 A food shine and / or gloss improver comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4.
  20. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を食品に添加することを含んでなる、風味が改善された食品の製造方法。 A method for producing a food with improved flavor, comprising adding the glucan according to any one of claims 1 to 3 or a reduced product thereof or the liquid sugar or powdered sugar according to claim 4 to the food. .
  21. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を、苦味および／または渋味を有する食品に添加することを含んでなる、苦味および／または渋味が低減された食品の製造方法。 Adding the glucan according to any one of claims 1 to 3 or a reduced product thereof or the liquid sugar or powdered sugar according to claim 4 to a food having a bitter and / or astringent taste, A method for producing a food with reduced bitterness and / or astringency.
  22. 　苦味および／または渋味を有する食品が、ポリフェノール類含有食品である、請求項２１に記載の苦味および／または渋味が低減された食品の製造方法。 The method for producing a food with reduced bitterness and / or astringency according to claim 21, wherein the food having a bitterness and / or astringency is a polyphenol-containing food.
  23. 　ポリフェノール類含有食品が、茶系飲料、果実飲料、炭酸飲料、野菜飲料、スポーツ飲料、乳性飲料、アルコール飲料、その他の飲料、アイスクリーム、ゼリー、ムース、飴菓子、ガム、フィリング、健康食品、またはサプリメントである、請求項２２に記載の苦味および／または渋味が低減された食品の製造方法。 Polyphenols-containing foods include tea-based beverages, fruit beverages, carbonated beverages, vegetable beverages, sports beverages, dairy beverages, alcoholic beverages, other beverages, ice creams, jellies, mousses, candy sweets, gums, fillings, health foods, The method for producing a food with reduced bitterness and / or astringency according to claim 22, which is a supplement.
  24. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を、医薬品に添加することを含んでなる、不快な味がマスキングされた医薬品の製造方法。 An unpleasant taste masked drug comprising adding the glucan according to any one of claims 1 to 3 or a reduced product thereof or the liquid sugar or powdered sugar according to claim 4 to the drug. Manufacturing method.
  25. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を食品に添加することを含んでなる、食感が改善された食品の製造方法。 Production of a food with improved texture, comprising adding the glucan according to any one of claims 1 to 3 or a reduced product thereof or the liquid sugar or powdered sugar according to claim 4 to the food. Method.
  26. 　添加物が均一に溶解または分散した水に、請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を添加して凍結させることを含んでなる、均一的に凍結した氷の製造方法。 The glucan or reduced product thereof according to any one of claims 1 to 3 or the liquid sugar or powdered sugar according to claim 4 is added to water in which the additive is uniformly dissolved or dispersed, and then frozen. A method for producing uniformly frozen ice comprising.
  27. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を食品に添加することを含んでなる、照りおよび／またはつやが向上した食品の製造方法。 A food with improved shine and / or gloss, comprising adding the glucan according to any one of claims 1 to 3 or a reduced product thereof or the liquid sugar or powdered sugar according to claim 4 to the food. Manufacturing method.
  28. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を、乳タンパク質含有飲料に添加することを含んでなる、乳タンパク質の凝集および／または沈殿が抑制された乳タンパク質含有飲料の製造方法。 Aggregation of milk protein comprising adding the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4 to a milk protein-containing beverage, and A method for producing a milk protein-containing beverage in which precipitation is suppressed.
  29. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる、食品。 A food comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4.
  30. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる、ポリフェノール類含有食品。 A polyphenol-containing food comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4.
  31. 　ポリフェノール類０．１３質量％に対して、請求項１～３のいずれか一項に記載のグルカンもしくはその還元物を１．６～５０質量％含有する、請求項３０に記載のポリフェノール類含有食品。 The polyphenol-containing food according to claim 30, wherein the glucan or reduced product thereof according to any one of claims 1 to 3 is contained in an amount of 1.6 to 50% by mass with respect to 0.13% by mass of the polyphenols. .
  32. 　ポリフェノール類含有食品が、茶系飲料、果実飲料、炭酸飲料、野菜飲料、スポーツ飲料、乳性飲料、アルコール飲料、その他の飲料、アイスクリーム、ゼリー、ムース、飴菓子、ガム、フィリング、健康食品、またはサプリメントである、請求項３１に記載のポリフェノール類含有食品。 Polyphenols-containing foods include tea-based beverages, fruit beverages, carbonated beverages, vegetable beverages, sports beverages, dairy beverages, alcoholic beverages, other beverages, ice creams, jellies, mousses, candy sweets, gums, fillings, health foods, Or the foodstuff containing polyphenols of Claim 31 which is a supplement.
  33. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる氷、またはそれを含有してなる食品。 An ice comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4, or a food containing the same.
  34. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる、医薬品。 A pharmaceutical comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4.
  35. 　請求項１～３のいずれか一項に記載のグルカンもしくはその還元物または請求項４に記載の液糖もしくは粉糖を含んでなる、乳タンパク質含有飲料。 A milk protein-containing beverage comprising the glucan according to any one of claims 1 to 3 or a reduced product thereof, or the liquid sugar or powdered sugar according to claim 4.
  36. 　経腸栄養剤または経口栄養剤である、請求項３５に記載の乳タンパク質含有飲料。 The milk protein-containing beverage according to claim 35, wherein the beverage is an enteral nutrient or an oral nutrient.
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