JP2006298763A - Method for producing nigerose-containing syrup crystal and crystal nigerose - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method which can obtain crystal nigerose and a syrup-containing crystal containing the same even from a relatively low purity nigerose. <P>SOLUTION: The method for producing a nigerose-containing syrup crystal comprises depositing crystal nigerose from a sugar solution containing ≥50 mass% nigerose based on the solid component and comprises a stirring step for starting crystallization and/or developing crystallization. The method for producing crystal nigerose comprises separating syrup from the nigerose-containing syrup crystal obtained by the above method. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a method for producing crystalline nigerose and honey-containing crystals containing the same.

  Nigerose was found in the acid hydrolyzate of Nigeran, a microbial polysaccharide derived from Aspergillus niger in the 1950s, and was found to have an α-1,3-glucoside bond with glucose as a constituent sugar. It is a disaccharide possessed by nature and hardly exists in nature [Non-patent Document 1].

  Nigerose was isolated from sake and koji soup in Japan and was called salmon biose, a non-fermentable sugar [Non-Patent Document 2]. As the name suggests, nigerose is found in fermented foods such as sake, mirin, miso, beer, and soup. In addition, the presence of honey and other beverages and foods we have taken on a daily basis has been confirmed, and it is known as an important carbohydrate that forms these rich and rich tastes.

  Nigero-oligosaccharides containing nigerose have a taste-improving function such as retention of umami under low salt and improvement of taste of high-intensity sweetener [Patent Document 1, Patent Document 2] and anthocyanins It is known to have a fading suppression effect of a system dye [Patent Document 3]. In other words, nigerooligosaccharides containing nigerose have a salty effect due to their unique taste, and at the same time, can maintain umami under low salt and enhance umami as well as due to the effects of temperature and light during storage. An anthocyanin pigment that easily fades has the effect of suppressing fading due to ultraviolet rays, sunlight, and temperature.

  As the tertiary function of nigerooligosaccharide containing nigerose, anti-cariogenic [Non-patent Document 3] and physiological activity of immunostimulatory / regulatory action have been reported [Patent Document 3]. The production methods of nigerose having such useful functions include organic synthesis [Non-patent Document 4], degradation of microbial polysaccharides erucinan and nigeran, transglycosylation with cyclodextrin glucanotransferase, or condensation with glucoamylase. A method using a reaction, a method capable of allowing acetic acid to act on a cyclic tetrasaccharide composed of α-1,3- and α-1,6-glucoside bonds [Non-patent Document 5] has been reported. To date, the most economical methods have been disclosed in which starch is used as a raw material and enzymatically utilizes a transglycosylation / condensation reaction [Patent Documents 4 and 5].

  However, nigerose obtained by the methods described therein is a liquid or amorphous powder. This amorphous powder has high hygroscopicity and is extremely difficult to maintain the powder state, and easily sticks and causes a so-called crying phenomenon, so that it is not stable as a powder. It was. For this reason, when such amorphous nigerose is used, it is necessary to use a desiccant in combination or to be cryopreserved.

On the other hand, it has been reported that dextran derived from Leuconostoc mesenteroides B strain was subjected to acetic acid decomposition, nigerose was separated by an activated carbon column, and then stored in 80% methanol to produce crystalline nigerose [Non-patent Document 6].
However, this report does not describe any crystallization conditions such as the purity and concentration of nigerose. Furthermore, after this report, there is no report that produced crystalline nigerose or a honey-containing crystal containing the same. Takeo and S.M. A report by Matsuzaki et al. States that nigerose was obtained as an amorphous powder and could not be crystallized [Non-patent Document 7]. Thus, a method for obtaining crystalline nigerose or honey-containing crystals containing the same has not been established at all, and industrial production has been impossible.
S. A. Barker, E .; J. et al. Bourne, M.M. Stacey, Studies of Aspergillus niger. Part I. The structure of the polyglucosane synthesized by Aspergillus niger 152, J. Org. Chem. Soc. , (1953) 3084-3090. Kazuo Matsuda, Kiyoshi Aso, Studies on non-fermentable sugars (IV) About unsolvable disaccharide in sake and rice bran soup, 31 (1953) 211-213. , Kazuo Matsuda, Goro Makishima, Kazuo Shibasaki, Kiyoshi Aso, Studies on non-fermentable sugars (XI) Unfermentable disaccharide in sake and rice bran soup (III), identification of sakebiose, nigerose and y-sugar , 32 (1954) 498-501. S. Imai, K .; Takeuchi, K. et al. Shibata, S .; Yoshikawa, S .; Kitahata, S .; Okada, S .; Araya, T .; Nishizawa, Screening of Sugers Inhibitory Against Sucrose-Dependent Synthesis and Adherence of Insoluble Glucan and Acid Production Products. Dent. Res. 63 (1984) 1293-1297. K. Takeo, S .; Kitamura, Y. et al. Murata, Synthesis of nigero-oligosacharides, Crabohydr. Res. , 224 (1992) 111-122. Abstracts of Annual Meeting of the Japanese Society for Agricultural Chemistry 2003 p. 68 F. Yamauchi, K .; Matsuda, Crystalline β-Nigerose, Nature, 204 (1964) 1088. Crabohydr. Res. , 113 (1983) 281-289. JP-A-10-210949 JP-A-10-234331 Japanese Patent No. 3364442 JP-A-9-299095 JP 2003-169665 A

  The present inventors have invented a method for obtaining crystalline nigerose and honey-containing crystals containing it by crystallization using nigerose with a high purity and a reduced content of contaminating disaccharides. (Japanese Patent Application No. 2005-033918). By this method, it has become possible to provide crystalline nigerose and honey-containing crystals containing the same, which were previously considered impossible to produce. However, since this method uses highly purified nigerose, a process for highly purifying nigerose is required. Therefore, there has been a demand for means capable of obtaining crystalline nigerose and honey-containing crystals containing the same even from relatively low-purity nigerose.

  An object of the present invention is to provide a method capable of obtaining crystalline nigerose and honey-containing crystals containing the same even from relatively low purity nigerose.

  As a result of intensive studies to achieve the above object, the present inventors have conducted stirring for crystallization and / or growth so that a sugar having a nigerose content of 50% by mass or more per solid content can be obtained. It was found that nigerose can be crystallized from the solution, and the present invention has been completed.

That is, the means for achieving the above object is as follows.
[1] A method for producing a nigerose-containing honey crystal in which crystalline nigerose is precipitated from a sugar solution containing 50% by mass or more of nigerose per solid content,
Said method comprising a stirring step for crystallization and / or growth.
[2] The method according to [1], wherein the sugar solution has a solid content concentration of 10% by mass or more.
[3] The method according to [1] or [2], wherein the solvent of the sugar solution is at least one selected from the group consisting of water, ethanol, and methanol.
[4] The method according to [3], wherein the solvent of the sugar solution is an aqueous methanol solution containing less than 80% by volume of methanol.
[5] The method according to any one of [1] to [4], wherein the crystal is precipitated in the presence of a seed crystal.
[6] The method according to any one of [1] to [5], comprising a cooling step for crystallization and / or growth.
[7] The method according to any one of [1] to [6], comprising powdering the nigerose-containing crystals.
[8] A method for producing crystalline nigerose, characterized in that crystalline nigerose is obtained by mashing from nigerose-containing honey crystals obtained by the method according to any one of [1] to [7].
[9] The method according to [8], comprising pulverizing the crystalline nigerose.

  According to the method of the present invention, crystalline nigerose and nigerose-containing honey crystals containing the same can be obtained from relatively low-purity nigerose. Furthermore, according to the method of the present invention, crystalline nigerose and honey-containing crystals containing it can be obtained from a relatively low concentration of the crystal mother liquor.

Hereinafter, the present invention will be described in more detail.

The method for producing a nigerose-containing crystal of the present invention is a method for producing a nigerose-containing crystal in which crystal nigerose is precipitated from a sugar solution containing 50% by mass or more of nigerose per solid content. It is characterized by including the stirring process for. Here, “nigerose-containing crystal” refers to a nectar-containing crystal containing crystalline nigerose.

  The raw material nigerose used in the present invention is not particularly limited, and can be obtained by a known method. Specifically, for example, a method of enzymatically utilizing a sugar transfer reaction / condensation reaction using starch having excellent economic efficiency as a raw material (see JP-A-9-299095 and Japanese Patent No. 3364442), α-1 , 3- and α-1,6-glucosidic cyclic tetrasaccharides can be used, and nigerose obtained by a method capable of allowing acetic acid to act (see Japanese Society for Agricultural Chemistry 2003 Annual Meeting Abstracts p.68) can be used. . Moreover, nigerose marketed as a reagent can be refine | purified and used as needed.

  In the present invention, the raw material nigerose is dissolved in an appropriate solvent to prepare a sugar solution. The sugar solution contains 50% by mass or more of nigerose per solid content. When nigerose prepared by the above-mentioned method or commercially available nigerose has a desired purity, it can be used as it is for preparing a sugar solution. Also, contaminating saccharides other than nigerose, especially contaminating disaccharides, are fractionated by ion chromatography, column chromatography using silica gel or activated carbon, fractionation by organic solvents such as alcohol and acetone, and separation by membranes with appropriate separation performance. The raw material nigerose can be highly purified by removal. Also, fermentation treatment with microorganisms that assimilate or decompose saccharides without using nigerose, such as lactic acid bacteria, acetic acid bacteria, yeast, etc., enzyme treatment with enzymes that decompose saccharides without breaking down nigerose, such as glucoamylase, etc. The raw material nigerose can also be highly purified by removing contaminating saccharides, particularly contaminating disaccharides, by seeds or two or more methods. The sugar composition can be determined by a known method such as high performance liquid chromatography.

  As described above, the sugar solution prepared using the raw material nigerose purified as necessary has a nigerose content per solid content of 50% by mass or more, preferably 60% by mass or more, more preferably 69 to 99. % By mass. The content of disaccharides other than nigerose in the raw material nigerose is 35% by mass or less, preferably 25% by mass or less, more preferably 0.5 to 20% by mass. When the nigerose content per solid content of the sugar solution is less than 50 mass, it is difficult to precipitate crystals.

  The sugar solution used for crystal precipitation can be prepared by dissolving the raw material nigerose in a suitable solvent. When dissolving the raw material nigerose, it is preferable to warm the solution. The heating temperature of the solution can be set as appropriate.

  In the present invention, since stirring is performed for crystallization and / or growth, the solid content concentration of the sugar solution is preferably determined in consideration of easiness of stirring. In order to facilitate crystallization, the sugar solution is preferably a supersaturated solution of nigerose. Considering the above points, the sugar solution is preferably a nigerose solution having a solid content concentration of 10% by mass or more, more preferably a nigerose solution having a solid content concentration of 15% by mass or more, and a solid content concentration of 25%. Even more preferred is a -55 wt% nigerose supersaturated solution.

  As the solvent of the sugar solution, at least one selected from the group consisting of water, ethanol, and methanol can be used, and an ethanol aqueous solution or a methanol aqueous solution is preferably used. The concentration of the aqueous ethanol solution and the aqueous methanol solution can be set as appropriate. In the present invention, for example, an aqueous ethanol solution containing 2% by volume or more, preferably 5 to 99% by volume of ethanol, or an aqueous methanol solution containing less than 80% by volume, preferably 2 to 79% by volume of methanol can be used.

  In the present invention, for example, the sugar solution is placed in an auxiliary crystal can at room temperature (for example, about 10 to 40 ° C.), and gradually cooled while stirring while controlling the temperature as necessary, thereby containing the nigrose containing crystalline nigerose. Crystals (nigerose-containing crystals) can be obtained as a mace kit. The temperature at the time of stirring can be a temperature at which the nigerose solution maintains the supersaturation degree, preferably a temperature in the range of 10 to 90 ° C. Examples of the stirring include (1) a method using a stirring blade, (2) a method using a magnetic stirrer, (3) a method of blowing air, and (4) rotating or reciprocally shaking the auxiliary crystal can itself. It can be performed by a known method such as a method, (5) a method of vibrating the auxiliary crystal can itself by ultrasonic vibration, or (6) a method of combining these methods. In any method, it is preferable to set the stirring conditions in consideration of the absence of foaming of the mother liquor, heat generation, coloring, modification, excessive oxidation, and the like.

  Further, the stirring speed is sufficient if the crystal mother liquor is sufficiently stirred, and the speed at which the crystallization rate is high and economically excellent can be appropriately set according to the size and shape of the container. That is, in order to sufficiently stir in a small container, a high stirring speed is required, and in a large container, a slow stirring speed is sufficient. The stirring speed is, for example, preferably in the range of 0.3 to 200 rpm, more preferably in the range of 0.5 to 170 rpm, and even more preferably in the range of 1 to 150 rpm. The stirring time can be determined while observing the state of crystallization, and can be, for example, 1 hour to 1 month, preferably 10 hours to 2 weeks.

  Crystal precipitation is preferably performed using a sugar solution containing crystalline nigerose or nigerose-containing honey crystals as seed crystals. The amount of the seed crystal can be adjusted as appropriate. In order to easily perform crystallization and / or growth from a sugar solution, it is preferable to cool the sugar solution. The cooling temperature can be set as appropriate.

  The obtained mace kit can be processed by a known method such as a honey method, a block pulverization method, a fluidized granulation method, or a spray drying method. Crystal nigerose can be obtained by subjecting the mace kit to a honey process to remove sugar components other than nigerose (honey). The honey can be performed by a known method. For example, crystal nigerose can be purified from nigerose-containing crystals by washing and drying the crystal component obtained by centrifuging the mace kit. The crystalline nigerose thus obtained can be further pulverized by, for example, the above-mentioned known method. Further, powdered nigellose, that is, nigerose-containing crystal powder containing crystal nigerose and a nectar liquid can be obtained by pulverizing the mase kit without carrying out the honey method. It is also possible to pulverize after adding other saccharides to the mace kit. In addition, after the mace kit is subjected to the honey process, other saccharides can be added to be powdered. The powdered nigerose thus obtained can contain 80% by mass or more of crystalline nigerose, and can contain honey liquor and / or saccharides other than nigerose as components other than crystalline nigerose. . Nectar liquid and / or other saccharides are glucose, fructose, trehalose, cordobiose, maltose, isomaltose, lactose, starch syrup, maltooligosaccharide, isomaltoligosaccharide, fructooligosaccharide, whey oligosaccharide, galactooligosaccharide, mannooligosaccharide, mannose It may contain at least one selected from the group consisting of containing hetero-oligosaccharide, dextrin, and sugar alcohol.

  According to the method of the present invention, crystalline nigerose having a purity of 85% or more, preferably 90 to 99.9%, and nigerose-containing honey crystals containing the same can be obtained. In addition, purity shall mean mass% here. Crystalline nigerose and nigerose-containing crystal powder containing it are less hygroscopic and easier to handle than amorphous nigerose powder. Low hygroscopicity means that when powdered nigerose is handled in a normal air-conditioned room, it does not stick easily and does not cause crying or deliquescence. More specifically, for example, when left in an environment of 25 ° C. and a relative humidity of 65%, it means that 7% or more of weight change or deliquescence does not occur.

  Crystalline nigerose and honey-containing crystals containing it can also be used in admixture with other sweetening ingredients. For example, various sugars such as sucrose, starch syrup, flour koji, glucose, fructose, maltose, isomerized sugar, lactose, honey, coupling sugar, fructosyl oligosaccharide, erythritol, sorbitol, maltitol, xylitol, mannitol, lactitol, One or two selected from sugar alcohols such as reduced xylo-oligosaccharides, reduced glucose syrup, aspartame, alitame, saccharin, glycyrrhizin, stevioside, rebaudioside, sucralose, acesulfame K and other sweeteners such as glycine and alanine It can be used in combination with more than one species. Further, if necessary, it can be used by mixing with a bulking agent such as dextrin and starch.

  Crystalline nigerose and honey-containing crystals containing it are non-toxic, harmless sweeteners and are white powders or solids with a unique sweetness that is rich, rich and rich. It is a stable carbohydrate with excellent heat resistance and acid resistance. It is colored appropriately by heating, and does not generate off-flavors when mixed and processed with other materials. It will damage other mixed materials. There is almost no. Also, osmotic pressure controllability, shaping, shining, moisture retention, viscosity, anti-caking property, anti-caking property, aroma retention, anti-discoloration, stability, anti-crystallization of other sugars, difficult It also has various properties such as fermentability, starch aging prevention, protein denaturation prevention, and lipid degradation prevention. Therefore, the crystalline nigerose and the nigerose-containing honey crystal containing the same, or mixed with other sweetening ingredients, are sweeteners, non-fermentable food materials, taste improvers, flavor improvers, quality improvers, It can be suitably used as a water separation inhibitor, a fragrance, a discoloration inhibitor, a lipid degradation inhibitor, a stabilizer, an excipient, a powdered substrate, and the like. Furthermore, crystalline nigerose and nigerose-containing crystals containing it are low in hygroscopicity and have adhesion and anti-caking properties even in powder form, so one or more of pullulan, hydroxyethyl starch, polyvinylpyrrolidone, etc. It can also be used as a tablet or a sugar-coated tablet in combination with these binders. For this reason, it can be advantageously used in various compositions such as various foods, foods, foods, feeds, feeds, cosmetics, and pharmaceuticals as it is or in combination with known materials as needed. As a known material, for example, a flavoring agent, a coloring agent, a flavoring agent, a reinforcing agent, an emulsifier, an antioxidant, an ultraviolet ray inhibitor, or a medicinal component can be used as appropriate.

  Furthermore, crystal nigerose and nigerose-containing crystals containing it have the effect of enhancing the salty taste and other umami tastes in addition to the effects of sweetening, taste improvement, flavor improvement and quality improvement of ordinary food and drink. Because it has, for example, soy sauce, powdered soy sauce, miso, powder miso, various sprinkles, mayonnaise, dressing, vinegar, three cups vinegar, powdered sushi vinegar, noodle soup, sauce, ketchup, grilled meat sauce, curry roux, stew element, Dashi-no-moto, various complex seasonings, mirin, new mirin, table top sugar, coffee sugar, various Chinese and other seasonings such as tsutsuyu, synthetic liquor, brewed sake, sake, fruit liquor, sparkling liquor, beer It can be effectively used for soft drinks such as coffee, cocoa, juice, carbonated beverages, lactic acid beverages and lactic acid bacteria beverages.

  In addition, it has a low-sweetness and very elegant sweetness, not only as sweetening, but also has an effect of improving sweetness and taste, and also has effects such as glazing, protein denaturation, and richness. Because it has rice crackers, hail, garlic sugar, rice cakes, potatoes, buns, fertilizers, potatoes, sheepskin, jelly, castella, jasper, bread, pie, crackers, biscuits, pudding, waffles, butter cream, custard Various food and drink such as cream, cream puff, sponge cake, donut, ice cream, sherbet, etc., pastes such as flower paste, peanut paste, fruit paste, various jams such as jam, marmalade, etc. Livestock paste products such as pickles, ham, sausage, salmon, chikuwa, fish paste products and surimi of raw materials, various delicacies It can be used to boiled or the like. In order to contain in the above food and drink, etc., in any manufacturing process of the food or drink, for example, mixing, kneading, dissolution, melting, soaking, infiltration, spraying, coating, spraying, pouring, crystallization, solidification A known method such as granulation can be selected as appropriate. In addition to the effects as described above, it also has effects such as prevention of crystallization of food and drink components and stabilization of enzymes such as amylase, glycosidase, lipase, and protease.

  Crystalline nigerose and honey-containing crystals containing it can improve the palatability of feed and feed for domestic animals, poultry, and other domestic animals such as bees, sharks, and fish, as well as the ability to stimulate immune and natural killer cells. Since it has properties such as QOL (Quality of Life) improving ability, it can also be used for purposes such as prevention of infectious diseases, nourishment, health maintenance and stress reduction. When used in the above feeds and feeds, grains such as corn, milo and barley, other carbohydrates such as glucose and starch, proteins derived from plants and animals, lysine, threonine and tryptophan amino acids, vitamin A It can be used by mixing with vitamins such as vitamin B2, vitamin B12, vitamin E, hormones, antibiotics and the like.

  Crystalline nigerose and honey-containing crystals containing it, when applied to the skin, like other natural carbohydrates, are hypoallergenic and effective in retaining moisture in the skin. be able to. For example, as cosmetics, oils and fats, alcohols, surfactants, pigments, fragrances, hormones, vitamins, animal extracts, plant extracts, microbial extracts, salts, UV absorbers, antioxidants, bactericides, antiperspirants, It can mix | blend with the 1 type (s) or 2 or more types suitably selected from a deodorizer, a refreshing agent, saccharides, a chelating agent, a thickener, etc.

  Crystalline nigerose and honey-containing crystals containing it can also be applied to pharmaceuticals such as infusion preparations and enteral nutrients. Crystalline nigerose and honey-containing crystals containing it have the same calories as crystalline glucose and crystalline maltose, immunostimulatory ability, natural killer cell activation ability, interleukin-12 production induction ability, QOL improvement ability, etc. Since it has properties, it can be used for nutritional supplementation, nutrition management, infection prevention, physical condition maintenance, and the like. When used in the above infusion preparations, other carbohydrates such as glucose, fructose, sorbitol and xylose, essential amino acids such as lysine, valine and isoleucine, amino acids such as alanine, arginine and proline, lipids and minerals Can be used by mixing with vitamins, trace elements, etc.

  As described above, nigerose has various properties such as immunostimulatory ability, natural killer cell activation ability, interleukin-12 production inducing ability, QOL improvement ability, and the like. It can be added to foods, pharmaceuticals, feeds, feeds, etc. for the purposes of management, nutrition and tonic, health maintenance, prevention of infectious diseases, physical condition maintenance and stress reduction. Since the crystalline nigerose, nigerose-containing crystals and powdered nigerose of the present invention have low hygroscopicity and are easy to handle, they can be suitably used for the preparation of foods, pharmaceuticals, feeds and feeds. Furthermore, since it has excellent storability and stability, it is suitable for applications that are used and distributed in a state in which crystallinity is maintained after preparation, such as tablets and powder drugs.

In addition, the following literature can be referred for the usage-amount in the case of using nigerose for the said objective, and its effect.
Japanese Patent No. 3396129, Japanese Patent Application Laid-Open No. 11-228425, Japanese Patent Application Laid-Open No. 2001-064174, Japanese Patent Application Laid-Open No. 2002-080364, Japanese Patent Application Laid-Open No. 2002-265366, Japanese Patent Application Laid-Open No. 2002-265385, Japanese Patent Application Laid-Open No. 2002-265385. 325555, Biosci Biotechnol Biochem 63: 373-8, 1999), Int Immunopharmacol 2: 151-9, 2002, Pharmacology and Treatment 29: 815-26, 2001, Pharmacology and Treatment 30: 81-90, 2002.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples. In the following, “%” means “% by mass” unless otherwise specified.
[Reference Example 1]
Preparation of nigerose (1)
Nigerose was prepared as described below using as a raw material a commercially available nigerooligosaccharide-containing syrup containing 40% or more of nigerooligosaccharide per solid content (eclipse taste oligo, manufactured by Nippon Shokuhin Kako Co., Ltd.). After inoculating baker's yeast in a solution of taste oligo in water so that the solid content is 7%, fermenting at room temperature for 24 hours, and assimilating glucose, maltose and maltotriose contained in taste oligo, Amylase was added, dextrin or maltooligosaccharide in the saccharified solution was decomposed to glucose and fermented as needed with yeast to obtain an aqueous solution containing 95% nigerooligosaccharide per solid content. The aqueous solution was decolorized with activated carbon and purified with an ion exchange resin, and then concentrated to a solid concentration of 50% to obtain syrup containing 95% of nigerooligosaccharide. This syrup was applied to an activated carbon column previously equilibrated with water, and after removing glucose, nigerose was eluted with a 2% ethanol solution (fraction A), and nigerose and other sugars were eluted with a 5% ethanol solution. (Fraction B). Eluted fractions A and B were collected, ethanol was removed by filtration and concentration operations, purified by ion exchange resin, and lyophilized to obtain 200 g of non-crystalline powder and 300 g of fraction B. The respective sugar compositions were fraction A (glucose 2%, nigerose 91%, trisaccharide 7%) and fraction B (glucose 2%, nigerose 72%, trisaccharide 26%). When diffraction X-rays of the powder of fraction A were measured by a powder X-ray diffraction method, diffraction was not shown as shown in FIG. The method for analyzing the sugar composition uses high performance liquid chromatography, and the analysis conditions are as follows.

Column: Shodex Asahipak NH2P-50 4E (4.6 mm ID × 250 mm)
Eluent: acetonitrile / water = 75/25
Flow rate: 0.9 ml / min
Detector: Differential refractometer Column temperature: 30 ° C

[Reference Example 2]
Preparation of nigerose (2)
Nigerose was prepared as described below, using as a raw material a commercially available nigerooligosaccharide-containing syrup (Nissan Taste Oligo, manufactured by Nippon Shokuhin Kako Co., Ltd.) containing 40 mass% or more of nigerooligosaccharide per solid content. After inoculating baker's yeast into a solution in which taste oligo is dissolved in water so that the solid content becomes 7% by mass, fermenting at room temperature for 24 hours, and assimilating glucose, maltose and maltotriose contained in taste oligo, Glucoamylase was added, and dextrin and maltooligosaccharide in the saccharified solution were decomposed to glucose and fermented as needed with yeast to obtain an aqueous solution containing 95 mass% of nigerooligosaccharide per solid content. This aqueous solution was decolorized with activated carbon and purified with an ion exchange resin, and then concentrated to a solid concentration of 50% by mass to obtain syrup containing 95% by mass of nigerooligosaccharide. The syrup was applied to an ion exchange column (resin, XFS-43278) that had been parallelized with water in advance, and a disaccharide fraction (glucose 11 mass%, nigerose 36 mass%, trisaccharide 53 mass%) was recovered (fraction X ). This was concentrated to a solid concentration of 50% by mass and applied to an activated carbon column previously equilibrated with water. After removing glucose, nigerose was eluted with a 5% by volume ethanol solution (fraction C). The eluted fraction C was collected, ethanol was removed by filtration and concentration, purified with an ion exchange resin, and freeze-dried to obtain 300 g of an amorphous powder. The sugar composition was fraction C (glucose 1% by mass, nigerose 86% by mass, trisaccharide 13% by mass). The method for analyzing the sugar composition was the same as in Reference Example 1.

[Reference Example 3]
Preparation of nigerose (3)
Fraction B and fraction C obtained in Reference Examples 1 and 2 were mixed at a weight ratio of 15:85, and 100 g of amorphous powder was prepared as fraction D. Further, 100 g of an amorphous powder in which fraction C and fraction X were mixed at a weight ratio of 50:50 was fractioned E, and amorphous powder in which fraction C and fraction X were mixed at a weight ratio of 70:30. 100 g of this powder was prepared as fraction F, and 100 g of amorphous powder prepared by mixing fraction C and fraction X in a weight ratio of 75:25 was prepared as fraction F. Each sugar composition consists of fraction D (glucose 1% by mass, nigerose 84% by mass, trisaccharide 15% by mass), fraction E (glucose 6% by mass, nigerose 61% by mass, trisaccharide 33% by mass), fraction. F (glucose 8% by mass, nigerose 51% by mass, trisaccharide 41% by mass) and fraction G (glucose 8.5% by mass, nigerose 48.5% by mass, trisaccharide 43% by mass). The method for analyzing the sugar composition was the same as in Reference Example 1.

[Example 1]
Powdered nigerose (fraction A; nigerose content 91%) obtained in Reference Example 1 was dissolved at a concentration of 14 to 50% by mass in a 78% by volume methanol solution while preparing 5 ml each. 2 mg each of crystalline nigerose was added to this mother liquor and left at 25 ° C. with stirring (150 rpm). When stirring was continued overnight, the mother liquor was found cloudy, and a maze kit that was completely clouded in 2.5 days was obtained. Each mace kit was separated by centrifuging, and the crystals were washed and dried to obtain a white powder. When these powders were observed with a polarizing microscope, a polarized image was observed, and it was confirmed to be a crystalline substance. Moreover, when these powders were analyzed under the same conditions as in Reference Example 1, the sugar composition was found to have a nigerose content of 95% by mass or more. The nigerose content in the honey solution was determined from the sugar composition, solid content, and mass of the honey solution obtained by mashing the mace kit by centrifugation, and the crystallization rate was calculated from the following formula. The results are shown in FIG. If the crystallization rate is 10% or more, it can be determined that crystals have been obtained.

[Examples 2-7, Comparative Example 1]
Powdered nigerose from fractions B to G were dissolved at a concentration shown in Table 1 while heating in a 78% by volume methanol solution to prepare 5 ml each. 2 mg each of crystalline nigerose was added to the mother liquor and left at 25 ° C. with stirring (200 rpm). When stirring was continued overnight, in Examples 2 to 7, the mother liquor was found cloudy, and a mace kit that was completely clouded in 2.5 days was obtained. On the other hand, in Comparative Example 1 in which the nigerose content per solid content in the sugar solution was 48.5% by mass, crystallization did not occur.
Each Mase kit obtained in Examples 2 to 7 was subjected to honey by centrifugation, and the crystals were washed and dried to obtain a white powder. When these powders were observed with a polarizing microscope, a polarized image was obtained, and it was confirmed to be a crystalline substance. Moreover, when these powders were analyzed under the same conditions as in Reference Example 1, the sugar composition was 90% by mass or more for the nigerose content. The crystallization rate was calculated in the same manner as in Example 1. The results are shown in Table 1.

[Example 8]
Powdered nigerose (fraction D; nigerose content 84%) was dissolved in a 75% by volume methanol solution while heating to prepare a crystal mother liquor having a solid content of 50% by mass and 35% by mass. The mixture was left at 25 ° C. with stirring at 50 rpm and 150 rpm, respectively. One week after the start of stirring, the mother liquor became cloudy, and a maze kit that became cloudy in 3 weeks was obtained. Each mace kit was separated by centrifuging, and the crystals were washed and dried to obtain a white powder. When these powders were observed with a polarizing microscope, a polarized image was obtained, and it was confirmed to be a crystalline substance. Moreover, when these powders were analyzed under the same conditions as in Reference Example 1, the sugar composition was 90% by mass or more for the nigerose content. The crystallization rate was calculated in the same manner as in Example 1. The results are shown in Table 2. From this result, it can be seen that the crystallization rate increases when the crystal mother liquor is sufficiently stirred.

  As described above, according to the present invention, crystalline nigerose and honey-containing crystals containing it can be obtained from nigerose having a low purity and a low concentration. According to the present invention, crystalline nigerose and honey-containing crystals containing it can be industrially produced in large quantities. In addition, amorphous powdered nigerose has high hygroscopicity and lacks storage stability and stability, whereas crystalline nigerose obtained by the present invention and honey-containing crystals containing it have low hygroscopicity. Excellent stability. Crystalline nigerose and honey-containing crystals containing it have low sweetness, rich body taste, moderate viscosity, and formability, sweetener, taste improver, flavor improver, quality improver, water separation inhibitor As a stabilizer, an excipient, a powdered substrate, etc., it can be effectively used in various foods, cosmetics, pharmaceuticals and other compositions.

The powder X-ray diffraction peak of the lyophilized powder fraction A (amorphous) is shown. The relationship between the crystal mother liquor concentration of nigerose and the crystallization rate is shown.

Claims (9)

A method for producing a nigerose-containing crystal comprising precipitating crystalline nigerose from a sugar solution containing 50% by mass or more of nigerose per solid content,
Said method comprising a stirring step for crystallization and / or growth. The method according to claim 1, wherein the sugar solution has a solid content concentration of 10% by mass or more. The method according to claim 1 or 2, wherein the solvent of the sugar solution is at least one selected from the group consisting of water, ethanol, and methanol. The method according to claim 3, wherein the solvent of the sugar solution is an aqueous methanol solution containing less than 80% by volume of methanol. The method according to claim 1, wherein the crystal is precipitated in the presence of a seed crystal. The method according to any one of claims 1 to 5, comprising a cooling step for crystallization and / or growth. The method according to any one of claims 1 to 6, comprising pulverizing the nigerose-containing honey crystals. A method for producing crystalline nigerose, characterized in that crystalline nigerose is obtained by mashing from a nigerose-containing crystal obtained by the method according to any one of claims 1 to 7. 9. The method of claim 8, comprising pulverizing the crystalline nigerose.