JP2013249479A - Molded article comprising branched starch - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a molded article having an improved storage stability, which comprises a novel starchy material having an anti-aging property.SOLUTION: A molded article is provided, which comprises a branched starch having a 6-α-maltosyl branched structure and/or a 6-α-maltotetraosyl branched structure and having a significant anti-aging property.

Description

  The present invention relates to a molded product containing a branched starch, and in particular, to a molded product containing a branched starch having a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure.

  Starch is a high molecular weight glucan stored mainly in cells of higher plant seeds and rhizomes, and is generally a mixture of amylose and amylopectin. Amylose is an α-1,4 glucan having a structure in which glucose is linearly bonded with α-1,4 bonds. On the other hand, amylopectin has a structure in which α-1,4 glucan having a degree of glucose polymerization of 6 or more is branched by α-1,6 linkages in the linear portion of α-1,4 glucan. Although starch swells and becomes viscous gelatinized starch when the aqueous dispersion is heated, it has the property of aging and causing gelation when allowed to cool. Starch has been gelatinized since long ago, and is used for food, and since it has excellent processability, low cost, and storability, it is used as the main ingredient of food, and further, thickener, water retention agent, Colloid stabilizers are also widely used for the purpose of improving the physical properties and maintaining the quality of foods. Further, starch is liquefied and industrially used as a raw material for glucose, isomerized sugar, maltooligosaccharide, chickenpox and the like. However, gelatinized starch and liquefied starch have problems such as aging during storage, easily causing gelation, loss of water retention and hardening, and deterioration in processing suitability.

  Under such circumstances, many attempts have been made to chemically modify the structure of starch and impart aging resistance to the gelatinized starch. Specifically, esterified starch, etherified starch, oxidized starch And various modified starches. However, modified starch is not preferred because it loses its original glucose structure by adding a special functional group or the like and there is a concern about safety in long-term use.

  In addition, proposals have been made to enzymatically modify starch to suppress aging. Patent Document 1 discloses a branching enzyme (branching enzyme; EC 2.4.1.) That cleaves α-1,4 bonds of starch and synthesizes α-1,6 bonds by transfer reaction. 18), a water-soluble macrocyclic structure formed by the action of 4-α-glucanotransferase (D-enzyme; EC 2.4.1.25) or CGTase (EC 2.4.1.19) A composition containing glucan has been proposed. However, the macrocyclic glucan has a problem in that the physical properties of the raw material liquefied starch are lost because the molecular weight is greatly lowered and the viscosity is lowered as the cyclic structure is formed. Further, Patent Document 2 uses a branching enzyme derived from Neurospora crassa and has a dense branch structure as compared with the starch of the raw material without reducing the molecular weight from gelatinized starch. A composition containing highly branched starch having a branched structure with a glucose polymerization degree of 4 to 7 as the center has been proposed. However, Neurospora crassa is a special mold and has not been industrially manufactured and used for industrial use because its safety has not been confirmed in producing foods and drinks. Further, Patent Document 3 uses a branching enzyme derived from barley (SBE-II) and phosphorylase, and has a branched structure centered on glucose polymerization degree 6 or 7, using glucose-1-phosphate and maltooligosaccharide as reaction substrates. There has been proposed a method of forming a branched starch having the following. However, it is extremely difficult to use barley-derived branching enzyme (SBE-II), phosphorylase, and the substrate glucose-1-phosphate for industrial production. Has not been made. Under such circumstances, physical properties such as aging properties were improved while maintaining the properties of texture, texture, function, etc. of the molded product when starchy material was blended, with almost no molecular weight reduction of starchy material. Therefore, it is desired to provide a molded article having excellent storage stability.

JP-A-8-134104 JP 2001-294601 A JP 2002-78497 A

  An object of the present invention is to provide a molded article having improved storage stability, which contains a novel starch having aging resistance.

  In order to solve the above-mentioned problems, the present inventors paid attention to the use of various glycosyltransferases, and in the process of earnestly researching, JP 2005-95148 A (Japanese Patent Application No. 2004-104) by the same applicant as the present invention. 174880)) cyclo {→ 6) -α-D-glucopyranosyl- (1 → 4) -α-D-glucopyranosyl- (1 → 6) -α-D-glucopyranosyl- (1 → 4) A cyclic maltosyl maltose having a structure of -α-D-glucopyranosyl- (1 →} (hereinafter abbreviated as “cyclic maltosyl maltose” in the present specification) from α-1,4-glucan Surprisingly, when a silmaltose-forming enzyme was allowed to act on a high concentration of liquefied starch or a partially decomposed product of starch, the formation of cyclic maltosyl maltose was slight, and the intermolecular and / or intramolecular It was found that the enzymatically modified starch was produced by the 6-α-maltosyl transfer action, and the structure and physical properties of the modified starch were examined. As a result, a new branched starch having a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure is produced from liquefied starch or a partially decomposed starch with little decrease in molecular weight or increase in reducing power. Furthermore, it was found that the branched starch having this 6-α-maltosyl branched structure and / or 6-α-maltotetraosyl branched structure has a remarkable aging resistance (Japanese Patent Application No. 2005). -298253 specification), and the molded product containing the branched starch has its texture, texture, function, etc., as compared with the case where ordinary starch is added. The present invention was completed by finding that it can be held stably for a long period of time.

  That is, the present invention provides a molded article containing a branched starch having a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure having remarkable aging resistance. Is a solution.

  According to the present invention, by using a novel branched starch having aging resistance, it is possible to suppress deterioration in quality caused by aging of starch in various molded products including chemical products and industrial products, and to stabilize the storage thereof. Can be improved.

It is a figure which shows the elution pattern in the gel filtration chromatography of the various branch starch obtained by making cyclic maltosyl maltose production | generation enzyme act on liquefied starch (waxy corn starch liquefaction liquid). It is the figure which compared the content of the malto-oligosaccharide whose glucose polymerization degree is 7 or less in the pullulanase digest of various branch starches obtained by making cyclic maltosyl maltose production enzyme act on liquefied starch (waxy corn starch liquefaction liquid). . It is a figure which shows the absorption spectrum of the iodine-starch complex of the various branched starch obtained by making cyclic maltosyl maltose production | generation enzyme act on liquefied starch (waxy corn starch liquefaction liquid). It is the figure which showed typically the structure of the raw material liquefied starch (waxy corn starch liquefied liquid) and the branched starch used by this invention. It is a photograph of a branched starch and raw material liquefied starch (waxy corn starch liquefied liquid) used in the present invention in a 25% concentration solution, dispensed into a glass test tube, and refrigerated at a temperature of 5 ° C. for 10 days.

The branched starch having a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure used in the present invention (hereinafter sometimes simply referred to as “branched starch” in the present specification). In the molecule, it means all starches having a structure branched by α-1,6 bonds with maltose units and / or maltotetraose units, and not only the inside of α-1,4 glucan chains in starches, Also included are those having a structure in which maltose and / or maltotetraose is linked with α-1,6 at the 6-position of the non-reducing terminal glucose. The molecular weight of the branched starch used in the present invention is not particularly limited, but is preferably 1.0 × 10 ⁴ daltons or more. The branched starch having a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure according to the present invention is a kind of starch debranching enzyme that specifically hydrolyzes α-1,6 bonds. When digested with a certain pullulanase, maltose per solid is 1.8 mass% or more and / or maltotetraose is 0.7 mass% (hereinafter, unless otherwise specified, mass% is expressed as “%”. ) The above is generated. Since the chain length (degree of glucose polymerization) of a normal starch generally has a peak at 9 to 10, it is extremely short and has a specific chain length in the branched starch used in the present invention. It has a branched structure and can be clearly distinguished from existing starch used as a raw material. Moreover, although this branch starch has a branch location increased as compared with normal starch and the straight chain portion is short, the molecular weight is hardly lowered.

  Further, in the branched starch having a 6-α-maltosyl branched structure and / or 6-α-maltotetraosyl branched structure used in the present invention, the number of branches branched by α-1,6 bonds is increased as compared with existing starches. 2,3,4-trimethyl-1,5 which shows the presence of glucose in which a 1-position and a 6-position hydroxyl group are involved in a glucoside bond in a partially methylated product by performing a known methylation analysis , 6-triacetylglucitol (hereinafter abbreviated as “2,3,4-trimethylated product”) is higher than that of the raw material starch, and is usually per solid of partially methylated product. It can be determined from showing 0.4% or more. Further, a degradation test (β-amylolysis) with β-amylase is performed, and the β-amylase degradation limit of the branched starch used in the present invention is smaller than that of the raw material starch.

  Furthermore, the branched starch having a 6-α-maltosyl branched structure and / or 6-α-maltotetraosyl branched structure used in the present invention is specifically described later in the experimental section. Even when it is made into a 25% aqueous solution and kept at 5 ° C. for 10 days, it does not substantially show white turbidity due to aging of starch, and has a characteristic of showing remarkable aging resistance compared to raw material liquefied starch. .

  As a method for producing a branched starch having a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure used in the present invention, for example, a liquefied starch is used as a raw material, and this acts on the starch. A method using an enzyme that generates a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure in the molecule is preferable. Such enzymes act on liquefied starch, recognize the maltose structure present at the non-reducing end, and recognize this maltose as the other non-reducing end glucose residue in the starch molecule or the 6th position of the glucose residue inside the starch molecule. Any enzyme can be used as long as it undergoes α-maltosyl transfer to a hydroxyl group or catalyzes a reaction of transferring this maltose to a 4-position hydroxyl group of other non-reducing terminal glucose residues of starch molecules. For example, the cyclic maltosyl maltose producing enzyme disclosed in JP-A-2005-95148 by the same applicant as the present applicant can be suitably used.

The cyclic maltosyl maltose production mechanism of the cyclic maltosyl maltose producing enzyme that can be used in the production of the branched starch used in the present invention is as follows.
1) Acts on α-1,4 glucan having a glucose polymerization degree of 3 or more as a substrate, and converts the maltosyl residue at the non-reducing end to 6 of the non-reducing end glucose residues of other α-1,4-glucan molecules. A 6-α-maltosyl-malto-oligosaccharide having a 6-α-maltosyl group at the non-reducing end and a glucose polymerization degree increased by 2 by catalyzing a 6-α-maltosyl transition between molecules that transfer to a hydroxyl group; A maltooligosaccharide having a degree of polymerization reduced by 2 is produced.
2) Furthermore, it acts on 6-α-maltosyl-malto-oligosaccharide and cyclizes by intramolecular α-maltosyl transition, and cyclo {→ 6) -α-D-glucopyranosyl- (1 → 4) -α-D- A cyclic maltosyl maltose having a structure of glucopyranosyl- (1 → 6) -α-D-glucopyranosyl- (1 → 4) -α-D-glucopyranosyl- (1 →}, and a maltooligosaccharide having a glucose polymerization degree reduced by 4 Generate.
3) This enzyme catalyzes a slight intermolecular 4-α-maltosyl transfer, and generates a few malto-oligosaccharides having a glucose polymerization degree increased by 2 and a malto-oligosaccharide having a glucose polymerization degree reduced by 2 from the maltooligosaccharide. To do.
Enzymes that catalyze the above reaction are included in the cyclic maltosyl maltose-producing enzyme that can be used in the production of the branched starch used in the present invention regardless of its source, form, crude enzyme, or purified enzyme.

  Although the cyclic maltosyl maltose-producing enzyme used in the present invention is not limited by its source, a preferable source is a microorganism, for example, Arthrobacter globiformis M6 (Independent Administrative Agency, National Institute of Advanced Industrial Science and Technology) The cyclic maltosyl maltose producing enzyme produced by the deposit center, deposit number FERM BP-8448) is preferably used. The microorganisms having the ability to produce cyclic maltosyl maltose producing enzyme include not only the above-mentioned bacteria but also mutants thereof, and other microorganisms including recombinant microorganisms having the ability to produce cyclic maltosyl maltose producing enzyme, and their Mutants and the like are also included.

  The cyclic maltosyl maltose-producing enzyme used in the production of the branched starch used in the present invention may be a purified enzyme or a crude enzyme as long as it can be used for the preparation of the branched starch, and is a free enzyme. Even an immobilized enzyme can be used. 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.

  The starch used as a raw material for producing the branched starch used in the present invention is, for example, ground starch such as corn starch, waxy corn starch, rice starch, and sticky rice starch, potato starch, sweet potato starch, tapioca starch, and waste starch. Starch and the like can be advantageously used industrially. Furthermore, amylose obtained from starch, amylopectin, a partially degraded starch, and the like can be used as a raw material. In producing this branched starch from starch, it is preferable to use the above-mentioned raw material starch after gelatinization and / or liquefaction. A known method can be adopted as the starch gelatinization / liquefaction method itself.

  For example, the method of allowing a cyclic maltosyl maltose producing enzyme to act on liquefied starch can be preferably carried out under the following conditions. First, the concentration of liquefied starch is usually preferably 10% to 45%. If the concentration of the liquefied starch is less than 10%, the cyclic maltosyl maltose-forming enzyme is likely to catalyze the intramolecular maltosyl transfer reaction, and cyclic maltosyl maltose is generated rather than the branched starch, resulting in a decrease in the yield of the branched starch. On the other hand, if it exceeds 45%, it is difficult to dissolve starch in water.

  In producing the branched starch used in the present invention, the cyclic maltosyl maltose producing enzyme is used in an amount of 0.01 to 10 units, preferably 0.02 to 1 unit per gram of liquefied starch solid. Is done. The term “one enzyme unit” as used herein refers to one unit of the amount of enzyme that produces 1 μmol of cyclic maltosyl maltose per minute under the conditions of the activity measurement method of cyclic maltosyl maltose producing enzyme described later. If the amount of cyclic maltosyl maltose-producing enzyme used is less than 0.01 unit, the reaction is insufficient and there is no meaning of adding the enzyme. On the other hand, if it exceeds 10 units, the effect reaches its peak and the production cost increases. Neither is preferred.

  The reaction temperature in the enzyme reaction may be up to the temperature at which the reaction proceeds, that is, up to about 60 ° C. Preferably, a temperature around 30 ° C. to 50 ° C. is used. The reaction pH is usually adjusted to a range of 5 to 9, preferably 5 to 7. The amount of enzyme used and the reaction time are closely related and may be appropriately selected depending on the progress of the target enzyme reaction.

  The reaction product obtained by the reaction can be directly blended into the molded product as a branched starch product. If necessary, the product obtained by the reaction is centrifuged, filtered or the like to remove insoluble matters, and the water-soluble fraction is concentrated to obtain a target branched starch solution. Although the obtained branched starch solution can be used as it is, it is optional for drying and use as a powder so as to be advantageous for storage and easy to use depending on the application. Drying can usually be performed by freeze drying, spray drying or drum drying. The dried product is desirably pulverized as necessary.

  Although the reaction product obtained by reacting cyclic maltosyl maltose-producing enzyme with liquefied starch usually contains a small amount of cyclic maltosyl maltose together with the branched starch, this reaction product is used as it is for the preparation of a molded product as a branched starch. Can be used. Further, if necessary, such oligosaccharide can be removed and used as a purified branched starch for the preparation of a molded product. As a purification method, a conventional polysaccharide purification method such as gel filtration chromatography may be appropriately selected as necessary.

  The branched starch thus obtained has a feature that even if the solution is left at a low temperature, white turbidity due to aging is not observed and remarkable aging resistance is observed as compared with ordinary starch. . In general, starch is insoluble in cold water, but the branched starch used in the present invention is soluble in cold water up to at least 20%. In addition, this branched starch has an aqueous solution having a low viscosity as compared with the raw starch liquefaction liquid, and is excellent in handleability.

  When the branched starch used in the present invention is used as a substitute for ordinary starch in a molded product containing starch, it itself has aging resistance, so that a molded product in which aging of starch is suppressed is obtained. Therefore, the molded product containing the branched starch of the present invention is one in which a decrease in water retention, shape retention, freezing resistance, digestibility and the like due to aging of the starch is suppressed. In addition, this branched starch is added to taste improvers, quality improvers, water separation inhibitors, stabilizers, discoloration inhibitors, excipients, inclusion agents, binders, adhesives, molding agents, shaping agents, thickeners. The molded product of the present invention, blended as an agent, stabilizer, etc., is provided with moderate viscosity, shine, moisture retention, aging resistance, freezing resistance, drying resistance, heat resistance, retention, etc. Stability is improved.

  The molded product as used in the present invention means food, drink, food, feed, pet food, cosmetics, quasi-drugs, pharmaceuticals, agricultural chemicals, civil engineering greening products, agricultural and forestry supplies, horticultural supplies, biocompatible medical supplies. It refers to materials and miscellaneous goods.

  The molded product containing the branched starch of the present invention is particularly suitable as a molded product requiring biodegradability and / or slow disintegration with respect to water containing hot water. Specifically, for example, paper, non-woven fabrics, woven fabrics, knitted fabrics, yarns, fibers including slit fibers, ropes, tubes, ropes, foamed containers, hamburgers, ice cream, ramen, juices, coffee, beer, milk, etc. Containers for food and ice cream cone cups, tableware, trays, dishes, cups, cartons, garbage bag containers, packaging boxes, agricultural and horticultural pots, artificial wood, foam sheets, films, capsules, rose cushions, Adhesive moldings, sheets for agricultural house sheets, construction and civil engineering sheets, various packaging films including agricultural films, and coating moldings such as paint, cement, concrete, plastic It can be advantageously used for chemical products such as and industrial products. There are no particular restrictions on the form of these molded products, and they may be solutions, semi-solids, solids, pastes, foams, or films, sheets, tubes, capsules, short bars, plates, chips. What was shape | molded etc. may be sufficient.

  More specifically, these molded products can be used in fields where plastic foams are conventionally used. In particular, the low foam is suitable for packaging materials for electric appliance cabinets, automobile handles, bumpers, interior parts, and the like. Other uses that require lightness and safety, such as home interior items, hotel toothbrushes, spoons for in-flight meals, forks, dishes and trays, toys, air gun balls, stationery, office supplies, etc. .

  Moreover, as a high foam, it is effective as an alternative material of a polystyrene foam which has a problem in disposal especially now. For example, food trays, tableware packaging containers such as instant noodle containers, shipping boxes for marine and agricultural products, packaging boxes, cushioning materials such as cushioning materials for electrical products and precision equipment, and architectural and road use The soundproofing and heat insulating material is suitable.

  In addition to the above, the molded product containing the branched starch of the present invention includes clothing such as hats, ponchos and windbreakers, packaging materials such as garbage bags and souvenir bags, exercise equipment such as ski poles, etc. Can do. Also, cards that are consumed in large quantities, such as telephone cards, orange cards, pachinko cards, book cards, etc., various credit cards, library use cards, various membership cards, etc. It is optional to use it.

  In order to further improve the water resistance, chemical resistance, heat resistance, mechanical strength, etc. of these molded products, it is optional to treat the surface of the molded products of the present invention with various materials. For example, a metal, such as aluminum, non-plastics, or other biodegradable plastics such as polylactic acid having a higher melting point, treatments such as coating, laminating, dipping, or vapor deposition are effective. In addition, when treating the surface with metals or non-biodegradable plastics, to prevent the biodegradability after use from being deteriorated, surface treatment is applied only to parts that require water resistance, etc. It is also optional to take measures such as partially leaving the processing part.

  Moreover, as a molded product containing the branched starch of the present invention, civil engineering planting supplies such as piles, piles, golf tees, agricultural films, seedling pots, agricultural and horticultural pots, agricultural and forestry supplies, horticultural supplies, etc. It is preferable to knead known fertilizers such as nitrogen, phosphorus, potash, etc., effective fungi, and / or agricultural chemicals in advance because they become more effective as fertilizers after biodegradation. The addition ratio is 0% to 80%, preferably 5% to 30%.

  As a method of incorporating the branched starch as described above into various molded products, it may be incorporated in the process until the product is completed, for example, mixing, kneading, dissolution, immersion, penetration, spraying, coating, coating Well-known methods such as spraying, pouring and solidifying are appropriately selected. The amount is usually 0.1% or more, preferably 1% or more, and more preferably 2% or more, and can be appropriately selected according to the purpose.

  The method for producing a molded product containing the branched starch of the present invention will be described more specifically. For example, by using an ordinary plastic molding machine, the molded product is formed into a desired form such as a film, a sheet, a tube, or a capsule. be able to. The molding method is not particularly limited. For example, an appropriate method such as extrusion molding, injection molding, pressure molding, mold molding, cast molding, blow molding, stamping molding, cutting molding, thermoforming, and film molding is used. be able to. The resulting molded product can be used as a biodegradable molded product.

  In the molded product containing the branched starch of the present invention, if necessary, the polymer material is water-soluble other than the branched starch having a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure. Polysaccharides such as starch, starch partial degradation products, amylose, amylopectin and other starch derivatives, esterified, etherified, oxidized and / or crosslinked starch derivatives, pullulan, sodium alginate, agar, pectin, xanthan gum, dextran , Mucopolysaccharides such as carrageenan, native gellan gum, galactomannan and chondroitin sulfate, polylactic acid and its derivatives, polyalginic acid and the like can also be used in combination. In addition, a plasticizer or a gelling agent may be advantageously added to adjust the plasticity of the molded product. Examples of the plasticizer include water and various polyols, for example, polyhydric alcohols such as glycerin and polyvinyl alcohol, sugar alcohols such as erythritol, xylitol, sorbitol, maltitol, α, α-trehalose, cyclodextrin, international publication WO 02 / Cyclo {→ 6) -α-D-glucopyranosyl- (1 → 3) -α-D-glucopyranosyl- (1 → 6) -α-D-glucopyranosyl- (1 → 3) disclosed in No. 10361 -Cyclic tetrasaccharide having a structure of α-D-glucopyranosyl- (1 →) (cyclic nigerosylnigelose), cyclic maltosyl maltose, cyclo disclosed in International Patent Application No. PCT / JP2005 / 17642 → 6)-[α-D-glucopyranosyl- (1 → 4)] cyclic pentasaccharide or cyclic hexasaccharide having a structure of n-α-D-glucopyranosyl- (1 →} (n means 4 or 5), carbohydrate derivatives of these cyclic oligosaccharides, etc. Non-reducing carbohydrates, urea, natural fats and oils such as soybean oil and castor oil, alkyl esters of organic acids, etc. In addition, thermosetting resins such as phenol resin, urea resin, melamine resin, epoxy resin, natural Mixed with powder or pellets of thermoplastic polymer materials such as rubber, shellac resin, polyethylene resin, polystyrene resin, polyvinyl chloride resin, polypropylene resin, acrylic resin, polyester resin, etc. Add a chemical agent, quality improver, preservative, etc., heat to 150 ° C to 250 ° C, and press mold to form a biodegradable starch-based plastic molding. It can also be advantageously carried it.

  Moreover, in addition to said component, this branch starch containing molded product can contain other inorganic and organic components suitably. Examples of the inorganic component include talc, titanium dioxide, calcium carbonate, sand, clay, limestone, diatomaceous earth, mica, glass, quartz, alumina, silica, glass, kaolin, and ceramics. Organic components include chitin, chitosan, collagen, fibroin, keratin, rosin, dammer, copal, starch, cellulose, wood flour, fiber, pulp, lignin, protein and its degradation products, wax, fats and oils, lipids, sugar fatty acid esters , Alcohols such as ethanol, sugars other than branched starch, cyclic saccharides, sugar alcohols, colorants, pigments, preservatives, flavoring agents, flavoring agents, binders, freshness-keeping agents, surfactants, builders, cobuilders, oxidation Examples thereof include an inhibitor, a bleaching agent, a brightening agent, a dispersant, an antifoaming agent, a water softener, an ultraviolet reflector, and an ultraviolet absorber. Furthermore, it is optional to add biologically active substances such as nutritional components, physiologically active substances, animal / plant extracts, enzymes, herbicides, fungicides, fungicides, antibiotics, insect repellents, repellents and the like.

  Hereinafter, the branched starch used in the present invention will be described more specifically by experiments.

<Experiment 1: Preparation of cyclic maltosyl maltose producing enzyme>
Prior to the experiment, Arthrobacter globiformis M6 (FERM BP-8448) was cultured, and the cyclic maltosyl maltose-producing enzyme in the culture supernatant was purified to prepare an enzyme preparation.

<Experiment 1-1: Culture of Arthrobacter globiformis M6>
Partially decomposed starch (trade name “Paindex # 4”, manufactured by Matsutani Chemical Industry Co., Ltd.) 1.5 w / v (mass / volume)%, yeast extract (trade name “Polypeptone”, manufactured by Nippon Pharmaceutical Co., Ltd.) 0 0.5 w / v%, yeast extract (trade name “Yeast Extract S”, manufactured by Nippon Pharmaceutical Co., Ltd.) 0.1 w / v%, dipotassium phosphate 0.1 w / v%, monosodium phosphate dihydrate 0.06 w / v% of product, 0.05 w / v% of magnesium sulfate heptahydrate, 0.3 w / v% of calcium carbonate, and water, 100 ml each were put into two 500 ml Erlenmeyer flasks, Sterilized in an autoclave at 121 ° C for 20 minutes, cooled, inoculated with Arthrobacter globiformis M6 (FERM BP-8448), and cultured at 27 ° C and 230 rpm for 48 hours with shaking as seed culture. . About 20 L of a liquid medium having the same composition as the seed culture is put in a fermenter having a capacity of 30 L, heat-sterilized and cooled to a temperature of 27 ° C. The culture was aerated and stirred for 96 hours while maintaining 8.0. After culturing, the culture solution was extracted from the fermenter, centrifuged (8,000 rpm, 20 minutes) to remove the cells, and about 18 L of culture supernatant was obtained.

  The enzymatic activity of the cyclic maltosyl maltose producing enzyme was measured by the following method. Soluble starch was dissolved in 50 mM acetate buffer (pH 6.0) containing 2 mM calcium chloride so as to have a concentration of 2 w / v% to obtain a substrate solution. 0.5 ml of the enzyme solution was added to 0.5 ml of the substrate solution, and The reaction solution was heated at about 100 ° C. for 10 minutes to stop the reaction, and then α-glucosidase (“transglucosidase L“ Amano ”was used to decompose the remaining soluble starch and contaminated oligosaccharides. ”, Amano Enzyme Co., Ltd.) 4000 units per gram of solids and glucoamylase (“ GlucoTeam ”, sold by Nagase Seikagaku Corporation) 250 units per gram of solids, added at 50 ° C. for 1 hour. The amount of cyclic maltosyl maltose in the treatment solution is quantified by a high performance liquid chromatography (hereinafter abbreviated as “HPLC”) method. One unit of activity of the cyclic maltosyl maltose producing enzyme is defined as the amount of enzyme that produces 1 μmol of cyclic maltosyl maltose per minute under the above conditions. HPLC is performed using “Shodex SUGAR KS-801” (manufactured by Showa Denko KK) for the column, water as the eluent, at a column temperature of 60 ° C. and a flow rate of 0.5 ml / min. This was performed using a differential refractometer RI-8012 (manufactured by Tosoh Corporation).

<Experiment 1-2: Purification of cyclic maltosyl maltose producing enzyme>
Ammonium sulfate was added to about 9.2 L of the culture supernatant to a final concentration of 60% saturation, and the mixture was allowed to stand at 4 ° C. for 24 hours for salting out. The produced salting-out precipitate was collected by centrifugation (11,000 rpm, 30 minutes), dissolved in 10 mM Tris-HCl buffer (pH 7.5), dialyzed against the same buffer, and crude enzyme. About 240 ml was obtained as a liquid. This crude enzyme solution was subjected to anion exchange chromatography (gel volume: 100 ml) using “DEAE-Toyopearl 650S” gel manufactured by Tosoh Corporation. Cyclic maltosyl maltose producing enzyme activity is adsorbed on “DEAE-Toyopearl 650S” gel equilibrated with 10 mM Tris-HCl buffer (pH 7.5) and eluted with a linear gradient from 0 M to 0.4 M in salt concentration. As a result, it was eluted at a salt concentration of about 0.22M. This active fraction was recovered, ammonium sulfate was added to a final concentration of 1 M, and the mixture was allowed to stand at 4 ° C. for 24 hours. After centrifugation, the insoluble matter was removed, and “Phenyl-Toyopearl” manufactured by Tosoh Corporation was used. ) 650M ”gel for hydrophobic chromatography (gel volume 10 ml). Cyclic maltosyl maltose producing enzyme activity is adsorbed on a “Phenyl-Toyopearl 650M” gel equilibrated with 20 mM acetate buffer (pH 6.0) containing 1M ammonium sulfate, and a linear gradient with an ammonium sulfate concentration of 1M to 0M. As a result, it was eluted at an ammonium sulfate concentration of about 0.1M. Table 1 shows the cyclic maltosyl maltose producing enzyme activity, the cyclic maltosyl maltose producing enzyme specific activity and the yield in each step of this purification.

  The purified sample of cyclic maltosyl maltose-producing enzyme after hydrophobic chromatography was subjected to 5-20 w / v% gradient polyacrylamide gel electrophoresis, and the purity of the enzyme preparation was tested. It was a high standard.

<Experiment 2: Preparation of Branched Starch>
In order to investigate the structure and physical properties of the reaction product produced when cyclic maltosyl maltose producing enzyme was allowed to act on liquefied starch, the following experiment was conducted.

<Experiment 2-1: Enzyme reaction>
2,500 g of commercially available waxy corn starch (sold by Sanwa Starch Co., Ltd.) was suspended in 25 L of tap water containing 1 mM calcium chloride and adjusted to pH 6.0 with 2N hydrochloric acid to prepare a 10% concentrated starch milk. . To this starch milk, 20,000 units of α-amylase (trade name “Neospirase PK2”, manufactured by Nagase Seikagaku Corporation) was added, stirred for 30 minutes, and then passed through the continuous liquefaction apparatus at a flow rate of 1 L / min. Liquefied starch (waxy corn starch liquefied liquid) was prepared by heating starch milk in a continuous liquefaction apparatus at 100 ° C. for 25 minutes and then at 140 ° C. for 5 minutes. The obtained liquefied liquid was decolorized with activated carbon, filtered through diatomaceous earth, and then concentrated under reduced pressure to a concentration of 25%. This concentrated liquefied liquid is divided into five equal parts, and the cyclic maltosyl maltose-producing enzyme purified preparation obtained in Experiment 1 is divided into four liquefied liquids at 0.0125, 0.025, 1 gram of waxy corn starch liquefied liquid solids. It added in the ratio of 0.05 or 0.1 unit, and was made to act at 50 degreeC and pH 6.0 for 24 hours. After stopping the enzyme reaction by heating at 100 ° C. for 10 minutes, the cyclic maltosyl maltose content in each reaction solution was measured by HPLC. In addition, the raw material liquefied starch which is not making cyclic maltosyl maltose production | generation enzyme act was made into the control | contrast.

  In addition, HPLC is performed using two “MCIgel CK04SS” columns (manufactured by Mitsubishi Chemical Corporation) connected in series, using water as the eluent, and a column temperature of 80 ° C. and a flow rate of 0.4 ml / min. The detection was performed using a differential refractometer RI-8012 (manufactured by Tosoh Corporation).

  As is clear from Table 2, the cyclic maltosyl maltose content tended to increase as the amount of cyclic maltosyl maltose producing enzyme increased in each reaction solution. However, the content was as low as about 5% even when 0.1 unit per gram of the solid content of the maximum enzyme action amount, waxy corn starch liquefied liquid examined this time, was applied.

<Experiment 2-2: Purification of Branched Starch>
Each reaction solution obtained in Experiment 2-1 was filtered, decolorized with activated carbon according to a conventional method, desalted with H-type and OH-type ion exchange resins and purified, and then concentrated to a solid content concentration of 20% with an evaporator. . Subsequently, in order to remove cyclic maltosyl maltose mixed as a by-product, column fractionation using a strongly acidic cation exchange resin (“Amberlite CR-1310”, Na type, manufactured by Organo Corporation) was performed. The resin was packed in four jacketed stainless steel columns with an inner diameter of 5.4 cm and connected in series to a total resin layer length of 240 cm. While maintaining the temperature in the column at 60 ° C., 5% v / v of waxy corn starch liquefied liquid was added to the resin, and 60 ° C. warm water was added at SV0.13 for fractionation, and the sugar composition of the eluate was determined by HPLC method. The polymer fraction without cyclic maltosyl maltose was collected. The obtained polymer fraction was concentrated to 25% and then vacuum-dried to obtain each branched starch powder in a yield of 90% or more per solid. These branched starches were subjected to the same HPLC analysis as in Experiment 2-1, and it was confirmed that no cyclic maltosyl maltose was contained.

<Experiment 3: Structural analysis of branched starch>
The following test was performed on the branched starch obtained by the method of Experiment 2, and the structure of the branched starch was examined.

<Experiment 3-1: Molecular weight distribution of branched starch>
The molecular weight distribution of the branched starch obtained by the method of Experiment 2 was examined by gel filtration analysis. Gel filtration analysis was performed by connecting two “TSK-GEL ALPHA-M” columns (manufactured by Tosoh Corporation) in series, using a 10 mM acid buffer (pH 7.0) as an eluent, a column temperature of 40 ° C., and a flow rate. Detection was performed under the condition of 0.3 ml / min, and detection was performed using a differential refractometer “RI-8012” (manufactured by Tosoh Corporation). The branched starch was dissolved in 10 mM acid buffer (pH 7.0) and subjected to membrane filtration as a sample for gel filtration analysis. As a control, waxy corn starch liquefied liquid (liquefied starch) before treatment with cyclic maltosyl maltose-producing enzyme was similarly analyzed. The molecular weight of glucan in the sample was calculated based on a molecular weight calibration curve prepared by subjecting a pullulan standard for molecular weight measurement (available from Hayashibara Biochemical Laboratories Co., Ltd.) to gel filtration analysis. The elution pattern in gel filtration chromatography is shown in FIG. In the following figures, a is a control (waxy corn starch liquefied liquid), and b, c, d and e are 0.0125 units and 0.025 cyclic maltosyl maltose producing enzyme per solid content of the liquefied liquid, respectively. It is a branched starch obtained by allowing the unit, 0.05 unit and 0.1 unit to act. Hereinafter, branched starches obtained by acting 0.0125 units, 0.025 units, 0.05 units and 0.1 units of cyclic maltosyl maltose producing enzyme per solid matter of the liquefied liquid are respectively used as branched starches 1 and 2. 3 and 4. In addition, the elution pattern of the gel filtration analysis of the branched starches 1, 2, 3, and 4 showed almost the same pattern when treated with any of the four types of enzyme units. 1 shows only the elution pattern of branched starch 4 (e in FIG. 1) treated with a) in 1 and the maximum amount of enzyme action (0.1 unit).

As is clear from FIG. 1, the control (a) waxy corn starch liquefied liquid showed one peak in gel filtration chromatography. The weight average molecular weight of glucan contained in this peak was calculated to be 1.1 × 10 ⁶ daltons from the calibration curve data. Elution in gel filtration chromatography of branched starch 4 (e in FIG. 1) obtained by allowing cyclic maltosyl maltose producing enzyme to act on 0.1 unit per gram of waxy corn starch liquefied liquid in waxy corn starch liquefied liquid The pattern was not much different from that of the control (a), and it was found that even when a cyclic maltosyl maltose producing enzyme was allowed to act, the molecular weight distribution did not change significantly.

<Experiment 3-2: Hydrolysis rate of branched starch>
The reducing power of the four kinds of branched starches 1, 2, 3, and 4 obtained in Experiment 2-2 or the control waxy corn starch liquefied liquid was measured. The total sugar content of each sample was measured by the anthrone-sulfuric acid method, and the reduced sugar content was improved by the Park Johnson method (Takusaku et al., “Carbohydrate Research”, Vol. 94, pages 205 to 213 (1981). The hydrolysis rate, which means the ratio (%) of the amount of reducing sugars in the total sugar amount, is expressed by the following formula: hydrolysis rate (%) = (reducing sugar amount / total sugar amount) × 100. The results are shown in Table 3.

  As is apparent from Table 3, the amount of action of the cyclic maltosyl maltose producing enzyme is 0.1 units per gram of waxy corn starch liquefied liquid solids, which is the largest, as compared with the control waxy corn starch liquefied liquid. The increase in hydrolysis rate was only about 0.1%, and the hydrolysis caused by the reaction of cyclic maltosyl maltose producing enzyme was almost negligible.

<Experiment 3-3: Methylation analysis of branched starch>
In order to examine the binding mode of glucose as a constituent sugar, the above four kinds of branched starches 1, 2, 3, and 4 obtained in Experiment 2-2 or the control waxy corn starch liquefied liquid were methylated according to a conventional method. Hydrolysis with acid, followed by reduction and acetylation, and the resulting partial methyl-acetylglucitol (hereinafter sometimes abbreviated as “partially methylated product”) is gas chromatographic method (hereinafter “GLC”). The composition of the partially methylated product was examined. The results are shown in Table 4.

  As is apparent from the results in Table 4, since no 2,3,4-trimethylated product was detected in the control waxy corn starch liquefied solution, glucose residues bound to other glucose at the 1-position and 6-position Is not considered to exist. On the other hand, in the branched starch obtained by allowing a cyclic maltosyl maltose producing enzyme to act, 2,3,4-trimethylated product was detected, and the ratio increased as the amount of action of the enzyme increased. This result indicates that as the amount of action of the enzyme increases, the structure in which another glucose is bonded to the 6-position of the glucose residue, that is, the branched structure increases. In addition, since the ratio of 2,3-dimethylated compounds is also increasing, the glucose residues bound to other glucose at the 1st, 4th and 6th positions are considered to be slightly increased. . This indicates that the 6-α-maltosyl transfer caused by the action of the cyclic maltosyl maltose producing enzyme is not only at the 6-position of the non-reducing terminal glucose residue of the substrate starch, but also at the glucose residue located inside the glucose chain constituting the starch. It also suggests that this occurs for the 6-position of the group.

<Experiment 3-4: Product in Pullulanase Digestion of Branched Starch>
For the purpose of investigating the branched structure of the above four kinds of branched starches 1, 2, 3 and 4 obtained by the method of Experiment 2-2 or the control waxy corn starch liquefied liquid, the α-1,6 bond of starch is specifically selected. Hydrolysis pullulanase (EC 3.2.1.41) was allowed to act, and the sugar composition of each pullulanase digest was examined.

  The above four kinds of branched starches 1, 2, 3, and 4 obtained by the method of Experiment 2-2 or the control waxy corn starch liquefied solution is dissolved in deionized water to a final concentration of 1 w / v%, and acetic acid is added. After adding a buffer solution (pH 6.0) to a final concentration of 20 mM, 20 units of pullulanase (reagent crystal product, manufactured by Hayashibara Biochemical Laboratories Co., Ltd.) is added per gram of substrate solid content, and the mixture is heated at 40 ° C. for 24 hours. Reacted. After the reaction, heat treatment was performed at 100 ° C. for 10 minutes to inactivate the pullulanase, and then the sugar composition of the reaction solution was measured by the same HPLC method as in Experiment 2. The results are shown in Table 5. Further, the results of the control waxy corn starch liquefied liquid and branched starch 3 (the amount of action of cyclic maltosyl maltose producing enzyme: 0.05 unit / g-waxy corn starch liquefied liquid) in Table 5 are plotted with the degree of polymerization of glucose on the horizontal axis. FIG. 2 is a graph showing the content (%) in the digest of pullulanase on the vertical axis. In addition, the code | symbol a and d in FIG. 2 means the control waxy corn starch liquefied liquid and the branched starch 3, respectively.

  As is apparent from the results in Table 5 and FIG. 2, in the case of the control waxy corn starch liquefied liquid, malto-oligosaccharides of DP6 or more were produced, and malto-oligosaccharides of DP5 or less were not observed, whereas cyclic maltosyl maltose was produced. In the branched starch prepared by the action of an enzyme, malto-oligosaccharides having a DP of 5 or less, particularly maltose (DP2) and maltotetraose (DP4) in the digest of pullulanase were remarkably increased. From this result, it was found that a new branched starch having a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure was generated by the action of the cyclic maltosyl maltose-producing enzyme.

<Experiment 3-5: β-amylase degradation limit of branched starch (β-amylolysis)>
The above four kinds of branched starches 1, 2, 3, and 4 obtained in Experiment 2-2, or the control waxy corn starch liquefied liquid, each with a final concentration of 1 w / v%, and acetate buffer (pH 5.5) with a final concentration of 20 mM. To the prepared substrate solution, 100 units of β-amylase (from soybean, manufactured by Nagase Seikagaku Corporation) per 1 g of solid matter was added, allowed to act at 50 ° C. for 24 hours, and heat-treated at 100 ° C. for 10 minutes for enzyme reaction. Stopped. One unit of β-amylase activity was defined as the amount of enzyme that produces a reducing power equivalent to 1 μmol of maltose per minute under the conditions of pH 5.5 and 40 ° C. with soluble starch having a concentration of 1% as a substrate. . The content of maltose and β-limit dextrin not degraded by β-amylase in the β-amylase digest of each branched starch and control waxy corn starch liquefaction solution was measured under the same HPLC conditions as in Experiment 1. The results are shown in Table 6.

  As is clear from the results in Table 6, the branched starch shows a lower maltose content produced by β-amylase digestion as the amount of the cyclic maltosyl maltose producing enzyme acted is larger. The content of dextrin was high. Since β-amylase is an enzyme that hydrolyzes starch in maltose units from the non-reducing end and stops the hydrolysis reaction just before the branching point due to α-1,6 bond, the above results indicate that cyclic maltosyl maltose is generated. The enzyme acts on the waxy corn starch liquefaction liquid, and a branched structure is formed by the α-1,6 maltosyl transition. The larger the amount of the cyclic maltosyl maltose producing enzyme, the higher the ratio of the branched structure in the branched starch (the branching is increased). Tells the story)

<Experiment 3-6: Iodine coloration of branched starch>
The above four kinds of branched starches 1, 2, 3, and 4 obtained in Experiment 2-2 or the control waxy corn starch liquefied liquid were dissolved in deionized water to a concentration of 0.15%, respectively. Add 10 ml of 0.02N sulfuric acid to 0.5 ml and 0.5 ml of 0.1N iodine-potassium iodide solution, leave it at 25 ° C. for 25 minutes, and then display the absorption spectrum of the iodine-starch complex in the range of 450 to 700 nm. It was measured. The results are shown in FIG. Note that the symbols a, b, c, d, and e in FIG. 3 are the same as those described in Experiment 3-1, FIG. As apparent from the results of FIG. 3, the branched starch having a larger amount of action of the cyclic maltosyl maltose producing enzyme has a lower absorbance as a whole. The maximum absorption wavelength was approximately 520 nm, and no difference was observed between the samples. From the results of Experiment 3-2, although the hydrolysis due to the action of the cyclic maltosyl maltose producing enzyme was hardly observed, the reason why the absorbance of the branched starch having a large amount of the action of the cyclic maltosyl maltose producing enzyme is lower, It was considered that the branched starch having a larger amount of cyclic maltosyl maltose-producing enzyme had a lower abundance ratio of the straight chain structure of starch forming a complex with iodine, and the amount of iodine bound to starch was reduced.

  From the results of Experiment 3, the branched starch obtained by reacting a liquefied starch (waxy corn starch liquefied liquid) with a cyclic maltosyl maltose-producing enzyme is a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched. It was found to be a novel branched starch having a structure. A schematic diagram showing the structure of this branched starch is shown in FIG. 4 together with that of liquefied starch (waxy corn starch liquefied liquid). In FIG. 4, A and B are schematic views of liquefied starch (waxy corn starch liquefied liquid) and branched starch used in the present invention, respectively. In the schematic diagram shown in FIG. 4, reference numerals 1, 2, and 3 denote a linear structure (amylose structure) in which glucose is linked by α-1,4 bonds in liquefied starch (waxy corn starch liquefied liquid), α, -1 and 6 represent a branched site of the linear structure and a reducing terminal glucose. Reference numerals 4 and 5 denote a 6-α-maltosyl branched structure and a 6-α-maltotetrao in branched starch. This means a sill branch structure.

<Experiment 4: Aging resistance of branched starch>
The above four types of branched starches 1, 2, 3, and 4 obtained by the method of Experiment 2 or the control waxy corn starch liquefied liquid were examined for aging resistance. Each sample heated and dissolved in water to a concentration of 25% was dispensed into a glass test tube and stored in a sealed state at a temperature of 5 ° C. for 10 days, and then the degree of starch aging was observed. The results are shown in FIG. In addition, the symbols a, b, c, d, and e in FIG. 5 are the same as those described in Experiment 3-1 and FIG. As is clear from the results of FIG. 5, the control waxy corn starch liquefied liquid (a) became cloudy under the above-mentioned storage conditions, and also partly solidified, and starch aging was remarkable. On the other hand, all of the branched starches (b to e) prepared by the action of a cyclic maltosyl maltose producing enzyme maintain a transparent solution state, and have a 6-α-maltosyl branched structure and / or 6-α-malto. It was found that the branched starch having a tetraosyl branched structure has remarkable aging resistance.

  Hereinafter, the present invention will be described more specifically with reference to examples. Production examples of the branched starch used in the present invention are shown in Examples 1 to 5, and molded articles containing the branched starch of the present invention are shown in Examples 6 to 24. However, the present invention is not limited to these examples.

  Waxy corn starch (manufactured by Sanwa Starch Kogyo Co., Ltd.) was suspended in tap water, and calcium chloride was added thereto so as to have a final concentration of 1 mM, and the pH was adjusted to 6.0 to prepare starch milk having a concentration of about 10%. To this starch milk, 0.05 mg of heat-resistant α-amylase (trade name “Spitase HS”, sold by Nagase Seikagaku Corporation) is added per gram of starch solid, stirred for 30 minutes, and then flowed to a continuous liquefaction device at a flow rate of 1 L. Per minute. The starch milk was heated in a continuous liquefaction apparatus at 100 ° C. for 25 minutes and then at 140 ° C. for 5 minutes to prepare liquefied starch. Next, this liquefied starch solution was concentrated under reduced pressure to obtain a liquefied starch solution having a concentration of about 25%, and then the cyclic maltosyl maltose-producing enzyme purified preparation obtained by the method of Experiment 1 was added to the 0.1% per gram of starch solids. The mixture was added so as to be a unit and reacted at pH 6.0 and a temperature of 50 ° C. for 20 hours. After stopping the enzyme reaction by heat treatment at 100 ° C. for 20 minutes, the filtrate obtained by cooling and filtration is decolorized with activated carbon and filtered through diatomaceous earth according to a conventional method, and a branched starch solution having a concentration of about 25% is solidified. The yield was about 90% per product. In addition, the pullulanase digest of the obtained branched starch contained 3.7% maltose and 1.7% maltotetraose. Moreover, the partially methylated product of the obtained branched starch contained 8.2% of 2,3,4-trimethylated product. This product contained 96.7% branched starch and 3.3% cyclic maltosyl maltose per solid. This product has moderate viscosity, moisture retention, aging resistance, and clathrate, taste improver, quality improver, water separation inhibitor, stabilizer, discoloration inhibitor, excipient, clathrate, As binders, adhesives, molding agents, shaping agents, thickeners, stabilizers, powdered base materials, various chemical products, industrial products, civil engineering greenery products, agricultural and forestry products, horticultural materials, food and drink It can be advantageously used for the production of various molded articles such as cosmetics, quasi drugs, pharmaceuticals, feeds, feeds, miscellaneous goods.

  The solution-like branched starch obtained in Example 1 was filtered, decolorized with activated carbon according to a conventional method, desalted with H-type and OH-type ion exchange resins and purified, and then concentrated with an evaporator to a solid content concentration of 20%. . Subsequently, it was subjected to column fractionation using a strongly acidic cation exchange resin (“Amberlite CR-1310”, Na type, manufactured by Organo Corporation) to remove cyclic maltosyl maltose mixed as a by-product. For the fractionation, the resin was packed in 4 jacketed stainless steel columns with an inner diameter of 5.4 cm and connected in series, and the resin layer had a total length of 240 cm. On the other hand, 5v / v% was added, and this was performed under the condition that warm water of 60 ° C was flowed at SV 0.13. A polymer fraction not containing cyclic maltosyl maltose was collected and concentrated to 25%, and then dehydrated, dried and powdered by a pulse combustion drying system PULCO (sold by Partec Co., Ltd.). By this operation, a branched starch powder having low hygroscopicity and excellent particle size characteristics was obtained. This product was a powder having a water content of about 10%, was easily dissolved in water up to a solid content concentration of 30%, and the water solubility was good. This product is a taste improver, quality improver, water separation preventive agent, stabilizer, discoloration preventive agent, excipient, inclusion agent, binder, adhesive, molding agent, shaping agent, thickener, stabilization Various chemicals, industrial products, food and drinks, civil engineering greenery products, agricultural and forestry products, horticultural materials, cosmetics, quasi-drugs, pharmaceuticals, feeds, feeds, miscellaneous products, etc. Can be advantageously used in the production of

  A commercially available starch partial degradation product (trade name “Paindex # 100”, sold by Matsutani Chemical Industry Co., Ltd.) is made into an aqueous solution with a concentration of about 30% (w / v), calcium chloride is added so that the final concentration is 1 mM, and pH 6. Adjusted to zero. To this was added 1 unit of gram of substrate-derived maltosyl maltose-producing enzyme obtained by the method of Experiment 1, reacted at 40 ° C. for 48 hours, heated to 100 ° C. and held for 10 minutes for reaction. Was stopped. The reaction solution was decolorized with activated carbon and purified by filtration through diatomaceous earth according to a conventional method, and further concentrated to obtain a branched starch partial decomposition product solution having a concentration of 30% at a yield of about 90% per solid. This product contains 90.8% partially degraded starch having a degree of glucose polymerization of 7 or more, 6.7% malto-oligosaccharides having a degree of glucose polymerization of 1 to 6, and 2.5% cyclic maltosyl maltose per solid. It was. This product has moderate viscosity, moisture retention, aging resistance, and inclusion properties, and is a taste improver, quality improver, water separation inhibitor, stabilizer, discoloration inhibitor, excipient, inclusion agent, and powder. For the production of various chemical products, industrial products, food and drink, cosmetics, civil engineering greenery products, agricultural and forestry supplies, horticultural supplies, quasi-drugs, pharmaceuticals, feeds, feeds, miscellaneous goods, etc. It can be used advantageously.

  In addition, when the raw starch partially decomposed product was digested with pullulanase, it was insolubilized due to aging of the produced amylose, whereas this product pullulanase digested product was resistant to aging and maintained a clear solution. In terms of conversion, it contained 26.3% of maltose and 15.4% of maltotetraose.

  When the cyclic maltosyl maltose-producing enzyme is allowed to act, the reaction is carried out in the same manner as in Example 3 except that 2,500 units of isoamylase is allowed to act per gram of substrate solids. The product was further concentrated to obtain a 30% -branched partial starch solution having a concentration of about 90% per solid. This product contains 69.6% partially decomposed starch having a glucose polymerization degree of 7 or more, 27.3% malto-oligosaccharide having a glucose polymerization degree of 1 to 6, and 3.1% cyclic maltosyl maltose per solid matter. It was. This product has moderate viscosity, moisture retention, aging resistance, and inclusion properties, and is a taste improver, quality improver, water separation inhibitor, stabilizer, discoloration inhibitor, excipient, inclusion agent, binding As chemicals, adhesives, molding agents, shaping agents, thickeners, stabilizers, powdered base materials, etc., various chemical products, industrial products, civil engineering greening products, agricultural and forestry products, horticultural materials, food and drink, It can be advantageously used for the production of various shaped articles such as cosmetics, quasi drugs, pharmaceuticals, feed, feed, sundries.

  The pullulanase digest of this product was a clear solution containing 41.5% maltose and 26.2% maltotetraose. When the maltose and maltotetraose content in the pullulanase digest of this product was compared with that of the pullulanase digest of the partially degraded starch obtained in Example 3, it was about 1.5 times higher. This means that when a cyclic maltosyl maltose producing enzyme is allowed to act while hydrolyzing a branch having a high degree of glucose polymerization with isoamylase, a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure Suggests that the number of can be increased.

  The solution-like branched starch partial decomposition product obtained in Example 4 was filtered, decolorized with activated carbon according to a conventional method, desalted with H-type and OH-type ion exchange resins, purified, and then evaporated to a solid content of 20%. Until concentrated. Subsequently, it was subjected to column fractionation using a strongly acidic cation exchange resin (“Amberlite CR-1310”, Na type, manufactured by Organo Corporation), and oligosaccharides containing mixed cyclic maltosyl maltose were removed. For the fractionation, the resin was packed in 4 jacketed stainless steel columns with an inner diameter of 5.4 cm and connected in series, and the resin layer had a total length of 240 cm. On the other hand, 5v / v% was added, and this was performed under the condition that warm water of 60 ° C was flowed at SV 0.13. A polymer fraction containing no oligosaccharide was collected, concentrated to 25%, dehydrated with a pulse combustion drying system PULCO (sold by Partec Co., Ltd.), dried and powdered. By this operation, a branched starch powder having low hygroscopicity and excellent particle size characteristics was obtained. This product was easily dissolved in water up to a solid content concentration of 30%, and its water solubility was good. This product is a taste improver, quality improver, water separation preventive agent, stabilizer, discoloration preventive agent, excipient, inclusion agent, binder, adhesive, molding agent, shaping agent, thickener, stabilization Various chemicals, industrial products, civil engineering greenery products, agricultural and forestry products, horticultural materials, food and drink, cosmetics, quasi-drugs, pharmaceuticals, feed, feed, sundries, etc. Can be advantageously used in the production of

<Carrot chip>
After washing, 150 g of carrot sliced into thin pieces of about 10 mm × about 10 mm × about 2 mm in thickness, α, α-trehalose (trade name “Treha”, sold by Hayashibara Corporation) and 10% of Example 1 It was immersed in a solution at about 100 ° C. containing 0.5% of the branched starch prepared by the method, and immersed for 15 minutes while maintaining the liquid temperature. The sliced carrot after the immersion treatment was drained, put into a hot air dryer, and dried in an atmosphere heated to 60 ° C. for 6 hours to produce a dried sliced carrot. This product is a carrot chip that is vivid, has good water return even when stored for a long period of time, and retains its carrot flavor.

  This product was pulverized by a conventional method to prepare a carrot powder. This powder is a carrot powder that retains the carrot flavor well even when stored for a long period of time, is colorful, does not solidify, and has high solubility in water.

<Green grass extract powder>
As a powdered base, 0.5 parts by mass of the branched starch prepared in Example 1 and α-cyclodextrin were added to 98 parts by mass of an aqueous extract of green grass (produced by Hayashibara Biochemical Research Institute, solid content: 1.4%). 1.5 parts by mass was added and stirred and dissolved, and this was spray-dried by a conventional method to prepare a cyanobacteria extract powder. This product is excellent in storage stability of useful components such as tryptanthrin and flavonoids contained in the water extract of cyanobacteria, even after long-term storage, without moisture absorption or browning. This product can be used as a raw material for producing foods, drinks, cosmetics, quasi drugs and the like.

  A chocolate containing 0.5% of this indigo extract powder was prepared by a conventional method. This product contains indigo extract and retains its useful components even after long-term storage, so it is used for the prevention of periodontal disease, prevention of hyperlipidemia, treatment and improvement of lipid metabolism. You can also.

<Royal jelly powder>
To 10 parts by weight of the royal jelly extract, 0.5 parts by weight of the branched starch prepared by the method of Example 1 and 1 part by weight of α, α-trehalose were added as a powdered base and stirred and dissolved. The powder was prepared by lyophilization by a conventional method. This product is excellent in the storage stability of useful components contained in royal jelly, even when stored for a long period of time, without moisture absorption or browning.

27 parts by mass of this royal jelly powder, 5 parts by mass of coenzyme Q _10, 5 parts by mass of sucrose, 55 parts by mass of erythritol, 6.5 parts by mass of ascorbic acid, 0.1 part by mass of vitamin B _1, 0.1 part by mass of vitamin B ₂ Vitamin B _{6 (} 0.1 part by mass) and fruit flavor (1.2 parts by mass) were mixed and subdivided into 1 g laminating containers. This product is suitable as a health supplement because its useful components are retained even after long-term storage.

<Powdered peppermint oil>
150 g of water, 70 parts by weight of gum arabic, 4 parts by weight of sucrose fatty acid ester, 2 parts by weight of powdered branched starch prepared by the method of Example 2 as a powdered base, and 20 parts by weight of hydrous crystals α, α-trehalose Was dissolved, heated to 85 ° C. for sterilization, and maintained for 15 minutes. After cooling to 40 ° C., 10 g of peppermint oil was added and emulsified with a homomixer. This was spray-dried by a conventional method to prepare powdered peppermint oil. This product did not absorb moisture, and the powder molded product maintained sufficient powder flowability. In addition, the oxidation and decomposition of peppermint oil is suppressed, not only the generation of deteriorated odor, but also the deterioration odor is masked even if deteriorated odor is generated, so the fragrance of good peppermint oil is stable Held for a long time. This product can be advantageously used as a fragrance for food and drink, cosmetics, quasi drugs, and pharmaceuticals.

  As a control, the peppermint powder prepared in the same manner as in Example 9 and the peppermint powder prepared in Example 9 except that only 20 parts by mass of hydrous crystals α, α-trehalose were used as the powdered base. Each container was placed in a sealed container and stored in a 50 ° C. incubator for 6 months. Using equal parts by weight of the preserved peppermint powder, each gum was prepared by a conventional method, and the flavor of the peppermint was subjected to a sensory test by 10 panelists. As a result, the peppermint powder prepared in Example 9 was used. Eight panelists evaluated that the gums using the scent had a stronger peppermint scent and higher persistence than the control. This result shows that the branched starch used as the powdered base contributes to the improvement of the storage stability of the fragrance.

<Powdered orange oil>
As a powdered base, 300 g of α, α-trehalose, cyclo {→ 6) -α-D-glucopyranosyl- (1 → 3) -α-D-glucopyranosyl- disclosed in WO 02/10361 (1 → 6) -α-D-glucopyranosyl- (1 → 3) -α-D-glucopyranosyl- (1 →} cyclic tetrasaccharide (produced by Hayashibara Biochemical Laboratories, Inc.) 300 g, α, α -A sugar derivative-containing sugar (trade name “Hellodex”, sold by Hayashibara Shoji Co., Ltd.) 40 parts by mass of trehalose and 30 g of the branched starch prepared by the method of Example 1 were dissolved in 200 g of ion-exchanged water. The mixture was heated while stirring at 100 rpm with a three-one motor to a boiling temperature of 135 ° C. 75 g of orange oil was added to the melt while stirring with a high-speed stirrer, and 20 minutes. The emulsified product was transferred to an extrusion kettle, pressurized into a cooling tank containing -25 ° C. isopropyl alcohol, extruded from the injection port, and pulverized while stirring. The isopropyl alcohol on the surface of the particles was removed by drying under reduced pressure at 40 ° C. The dried powder molded product was passed through a sieve of 20 mesh (aperture 840 μm) and 60 mesh (aperture 250 μm). Sieving was performed so as to remain on the sieve to obtain 480 g of a powder molded product remaining on the 60 mesh sieve.This product did not deteriorate perfume even after long-term storage, and the powder molded product had sufficient powder flowability. This product can be advantageously used as a fragrance for food and drink, cosmetics, quasi drugs, and pharmaceuticals.

<Spinning>
1.5 liters of an aqueous solution containing 8% polyvinyl alcohol having an average degree of polymerization of 1150 and a degree of saponification of 99.95%, and 2% of branched starch prepared by the method of Example 2, and reactive dye Kayacion
After 1.5 liters of 1% aqueous solution of Red E-SN7B (manufactured by Nippon Kayaku Co., Ltd.) was mixed and the pH was adjusted to 8 with caustic soda, the mixture was heated and reacted. Further, polyvinyl alcohol having an average polymerization degree of 1150 and a saponification degree of 99.95% and a branched starch and water prepared by the method of Example 2 were added to this colored aqueous solution, and the polyvinyl alcohol was 26% by the method of Example 2. A spinning dope containing 6% of the prepared branched starch and 0.3% of the dye was prepared. This spinning dope was dry-spun using a nozzle with 50 holes, drawn 4.5 times, and heat-treated at 220 ° C. to obtain a red colored yarn having a melting temperature of 93 ° C. This product has sufficient tensile strength and is excellent in durability.

<Lamination of polylactic acid film using branched starch>
A 5% aqueous solution of branched starch prepared by the method of Example 2 was applied onto a 2.5 μm thick polylactic acid film using an applicator and then dried to prepare a 3 μm thick branched starch / cast film. A dampened paper was placed on the branched starch surface of the obtained laminate film and dried while pressing. As a result, a three-layer laminate of a paper and a polylactic acid film using a branched starch as an adhesive layer was obtained. In order to measure the adhesive strength, a 90-degree peel test was performed. The adhesive strength was extremely excellent, and breakage occurred in the paper layer. However, the breakage in the branched starch layer, the branched starch layer and the paper or Neither interface delamination with the polylactic acid film occurred. In addition, since this product is biodegradable, it is an environmentally friendly film.

<Printed coating layer molding>
A pigment slurry comprising 50 parts by mass of heavy calcium carbonate and 50 parts by mass of kaolin was dispersed using a Coreless disperser to obtain a pigment slurry. In this slurry, 10 parts by mass (solid content) of styrene-butadiene copolymer latex, 3 parts by mass (solid content) of the branched starch liquid prepared in Example 1, 0.2 parts by mass of a water-resistant agent, and 0.2 parts by mass of a lubricant. Other additives were added and dispersed to prepare a paint having a solid content of 58%.

Using the coating material, double-side coating was performed with a blade coater so that the dry weight per side of the base paper was 20 g / m ² . After the paper was dried, water was supplied with a roll-up licator, water-coated, and dried. Thereafter, the surface was gloss-finished by super calendering to obtain a coated paper for printing. This product had little bleeding of printing ink and pigment, and had sufficient mechanical strength and durability.

<Paper>
Hardwood bleached kraft pulp was beaten with a Niagara beater and an appropriate amount was added to water to prepare a 400 ml suspension. Branched starch prepared by the method of Example 1 was mixed with the pulp as a powder, and hand-made by a paper sheet machine having a basis weight of 100 g / m ² according to a conventional method. The formed wet paper was dried with a rotary dryer at 90 ° C. for 1 minute and then conditioned at a temperature of 20 ° C. and a humidity of 65% for 24 hours to obtain a kraft paper. This product had sufficient mechanical strength and durability.

<Nonwoven fabric>
Hydrolysis copolymer having a saponification degree of 92% obtained by saponifying 60 parts by mass of a branched starch powder having a water content of about 10% prepared by the method of Example 2 and 30 mol% ethylene and 70 mol% vinyl acetate. A pellet of a biodegradable resin molded product comprising 40 parts by mass of a polymer was prepared. The pellet was melt-spun at a spinning temperature of 140 ° C. using a full flight screw of 0.8 mm, 350 holes and a compression ratio of 2.0 to obtain a regular yarn. To this yarn, 0.3% of the yarn mass was adhered to this yarn with potassium lauryl phosphate as a surface finish. The undrawn yarn was cold drawn at a draw ratio of 1.2 and then cut with a cutter to obtain a biodegradable fiber having a single yarn fineness of 6 d / f and a fiber length of 38 mm. This biodegradable fiber was carded with a card machine to obtain a card web. This web was further processed into a nonwoven fabric with an embossing roll at a temperature of 130 ° C. to obtain a nonwoven fabric. This product also had sufficient water resistance, mechanical strength and durability.

<Gardening sheet>
35 parts by weight of branched starch prepared by the method of Example 2 containing about 10% of water, copolymer of 30 mol% of ethylene and 70 mol% of vinyl acetate (partially hydrolyzed copolymer having a saponification degree of 98%) 60 Mass parts and 5 parts by mass of polycaprolactone were blended to obtain a biodegradable resin molded product. This was used for fiberization. After making this fiber into a web, the sheet | seat was obtained by the needle punch nonwoven fabric method / through-air process. This product can be used as a gardening sheet. This product had high water resistance and sufficient mechanical strength and durability. In addition, since this product is biodegradable, it is an environmentally friendly sheet that decomposes naturally even if left in a field after use.

<Antimicrobial sheet>
Carbohydrate containing α, α-trehalose-containing carbohydrate derivative (trade name “Hellodex”, Hayashibara Shoji Co., Ltd.) 45 parts by mass, sodium lactate 5 parts by mass, water 30 parts by mass, branch prepared by the method of Example 1 A coating solution was prepared by adding 20 parts by mass of starch. This coating solution was applied to kitchen paper so that the solid content was 0.8% with respect to the mass of the paper. This product can be used as an antibacterial sheet for packaging fresh food. Since this coating liquid contains the branched starch, the coating property to kitchen paper improves and it can apply | coat a coating liquid on paper uniformly.

<Sheet>
40 parts by mass of ethylene glycol and 1.4 parts by mass of Methbrene P530A were added to 100 parts by mass of the branched starch prepared by the method of Example 1, mixed at 1000 rpm for 10 minutes with a Henschel mixer, and then at 150 ° C. with a test extruder. Pelletized. After 100 parts by mass of the pelletized plastic molded product and 100 parts by mass of Bionore (# 1001) were tumbled, they were subjected to an extruder again at 150 ° C. to obtain a plasticized starch / bionore composite molded product. Using this molded product, a sheet having a meat pressure of 1 mm was formed with a T-die extruder under a heating temperature of 170 ° C.

  When a seedling pot was prepared by vacuum forming using this sheet, a beautiful molded product (seedling pot) could be obtained. This product also had high water resistance and sufficient mechanical strength and durability. In addition, since this product is biodegradable, it is an environmentally friendly sheet.

<Foamed material>
After mixing 100 parts by weight of the branched starch prepared by the method of Example 4, 1 part by weight of a copolymer of methyl methacrylate and alkyl acrylate, (molecular weight 3.1 million), 30 parts by weight of water with a Henschel mixer at 1000 rpm for 10 minutes. Then, it was pelletized at 100 ° C. with a test extruder (Toyo Seiki Co., Ltd., “Labo Plast Mill”). This was dried to a moisture of 13.5%, and a foam was formed at 190 ° C. using the pellets in a rose-shaped cushioning material manufacturing facility. This product replaces the branched starch used in the foamed material with the above composition, and provides a uniform foaming as compared to the foamed material prepared using starch, as well as resilience, foamability, moisture absorption, and water resistance. It is also excellent in physical properties such as properties, shape retention and durability, and can be advantageously used as a cushioning material. In addition, since this product is biodegradable, it is an environmentally friendly foam material.

<Tray>
100 parts by weight of branched starch (solid content) prepared by the method of Example 1, 20 parts by weight of water, 15 parts by weight of polyethylene glycol, and 0.04 parts by weight of potassium persulfate were placed in a Henschel mixer (sold by Mitsui Miike Chemical Industries) and 600 rpm Stir for 5 minutes and mix. This was pelletized with a lab plast mill type twin screw extruder and a pelletizer (both sold by Toyo Seiki Co., Ltd.). The pellets were injection molded using a molding die using an injection molding machine (sold by Nissei Plastic Engineering Co., Ltd.) to obtain a tray. This product has sufficient mechanical strength and is excellent in shape retention and water resistance, and can be used as a tray for plant cultivation. In addition, because this product is biodegradable, it is an environmentally friendly tray.

<Tray and pile>
A mixture of 47 parts by mass of aliphatic polyester (Bonore 1020, Showa Polymer Co., Ltd.), 47 parts by mass of branched starch prepared by the method of Example 1 and 6 parts by mass of sucrose (sugar) was supplied to an injection molding machine and molded. A molded product was obtained using a tray and a pile mold.

  The appearance of the obtained trays and piles was good, and there was no occurrence of kogation or discoloration due to the added sucrose. Moreover, the mechanical strength as a tray and a pile was sufficient, and washing with water was also possible. These moldings were biodegradable and were almost completely decomposed when buried in soil and allowed to stand at room temperature for 3 months.

<Emulsion>
Based on the following formulation, an emulsion was prepared by a conventional method.
(Prescription) (%)
Polyoxyethylene (20E.O.) polyoxypropylene (2E.O.) cetyl alcohol 1
Silicon KF96 (20cs) (Shin-Etsu Chemical Co., Ltd. sales) 2
Liquid paraffin (medium viscosity) 3
1,3-butylene glycol (1,3-BG) 5
L-ascorbic acid-2-glucoside 2
Glycerin 2
Branched starch prepared by the method of Example 5 0.6
Ethyl alcohol 15
Carboxyvinyl polymer 0.3
Hydroxypropyl cellulose 0.1
2-Aminomethylpropanol 0.1
Indigo extract (trade name “Indigo Luros”, sold by Hayashibara Biochemical Laboratories, Inc. Water extract of indigo plant containing 30% 1,3-BG) 2
Preservative Appropriate amount Purified water is added to make the total amount 100%.

  This product is useful as a skin external preparation for whitening and / or beautifying skin. In addition, this product is excellent in moisture retention, permeability, spreadability, and usability.

<Shampoo>
A shampoo was prepared by a conventional method based on the following formulation.
(Prescription) (%)
Piroctone Olamine 0.5
Edetate disodium 0.3
Photosensitive Element 201 0.002
Citric acid 0.3
Sodium salicylate 0.2
1,3-BG 3
Sodium polyoxyethylene lauryl ether sulfate 6.75
Coconut oil fatty acid diethanolamide 2
Lauryl sulfate triethanolamine 10
Polyoxyethylene lanolinic acid (80 EO) 0.5
2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine 10
Hydroxyethylcellulose hydroxypropyltrimethyl ammonium chloride ether 0.8
Branched starch prepared by the method of Example 2 0.5
Cyanobacteria extract powder prepared by the method of Example 7
Fragrance 0.2
Add purified water to bring the total volume to 100%.
This product has the effect of preventing inflammation and aging of the scalp and has strong antibacterial properties, so it has excellent hair growth effect, suppresses hair loss and keeps the scalp clean. Moreover, it is a shampoo excellent in feeling of use that retains moderate moisture after use and improves hair slipping.

<Hair restorer>
2 g powder of cyanobacteria extract prepared by the method of Example 7
Photosensitive Element 301 0.005g
Assembly extract 3.0ml
0.1g dipotassium glycyrrhizinate
α, α-trehalose 0.01 g
Seaweed extract 0.75g
Glycerin 2g
Purified water is added and stirred and dissolved to make the total volume 100 ml.
This product is a hair restorer that is excellent in hair growth effect and moisture retention, suppresses dandruff, itchiness and hair loss, and has a good feeling of use with good hair slippage.

  Since the branched starch used in the present invention has a dense branch structure and is resistant to aging, it is added to a starch-containing molded product as a starch substitute or a molded product used as a powdered base. The various quality deterioration resulting from aging of starch is reduced. Furthermore, the molded products include not only foods and drinks, cosmetics, quasi-drugs, feeds and feeds, but also chemicals such as sheets, fibers, foamed molded products, adhesives, industrial products, civil engineering greening products, agriculture and forestry. Since it can be used as a product, a horticultural material product, a powder product, a miscellaneous product, a biodegradable and / or slowly-decreasing molded product, the significance of the molded product containing the branched starch of the present invention in each industrial field is Extremely expensive.

a: Control liquefied starch (waxy corn starch liquefied liquid)
b: Branched starch 1 (cyclic maltosyl maltose producing enzyme acting amount 0.0125 unit)
c: Branched starch 2 (cyclic maltosyl maltose producing enzyme acting amount 0.025 unit)
d: Branched starch 3 (cyclic maltosyl maltose producing enzyme acting amount 0.05 unit)
e: Branched starch 4 (cyclic maltosyl maltose producing enzyme working amount 0.1 unit)
4A: Schematic diagram of liquefied starch B: Schematic diagram of branched starch used in the present invention 1: Chain structure in which glucose is linked by α-1,4 bonds (amylose structure)
2: Branched portion of chain structure by α-1,6 bond 3: Reducing terminal glucose 4: 6-α-maltosyl branched structure 5: 6-α-maltotetraosyl branched structure

Claims (2)

A quality improver comprising a branched starch having a 6-α-maltosyl branched structure and / or a 6-α-maltotetraosyl branched structure. The quality improving agent according to claim 1, wherein the quality of the molded product is improved by imparting aging resistance to the molded product containing starch.