JP2011200225A - Saccharide composition and food and drink - Google Patents

Abstract

The present invention provides a sugar composition that does not impair the flavor of food.
The sugar composition of the present invention is a sugar composition containing dietary fiber having glucose as a constituent sugar, and the numerical values of x, y and z in the following formulas are 0.20 ≦ x ≦ 0.75 and 0.25, respectively. A sugar composition in which ≦ y ≦ 0.80 and z ≦ 0.30.
x: Content of dietary fiber with a degree of polymerization of 3 and 4 with glucose as a constituent sugar (g) / Total content of dietary fiber with glucose as a constituent sugar (g)
y: Content of dietary fiber with a degree of polymerization of 5 to 9 using glucose as a constituent sugar (g) / Total content of dietary fiber containing glucose as a constituent sugar (g)
z: Content of dietary fiber with a degree of polymerization of 10 or more with glucose as a constituent sugar (g) / Total content of dietary fiber with glucose as a constituent sugar (g)
Such a sugar composition does not impair the flavor of the food when added to the food.
[Selection figure] None

Description

  The present invention relates to a sugar composition and food and drink using the same. More specifically, the present invention relates to a sugar composition containing dietary fiber and a food or drink using the same.

Conventionally, sugar compositions such as indigestible dextrin and polydextrose have been widely used in the food field such as beverages and confectionery as water-soluble dietary fibers.
Indigestible dextrin is produced by roasting starch (such as corn starch) in the presence of an acid. In the process of starch production, during starch roasting, intramolecular dehydration is performed with the reducing group of the glucose residue at the reducing end, or the dissociated glucose residue is randomly transferred to other hydroxyl groups. In addition to these bonds, 1, 2, 1, and 3 glucoside bonds are generated in the molecule (see Non-Patent Document 1 below). Although it is a sugar composition that can be produced at a low cost, there is a drawback that when added to food, the “crisp” of the taste of the food is canceled and the taste of the food is blurred (see Patent Document 1 below). Indigestible dextrin is a dextrin having a 1-6 anhydroglucose structure in which the glucose residue at the reducing end is dehydrated in the molecule, and has a unique flavor that occurs during the roasting process. There is also a problem that it affects.

  Polydextrose is produced by subjecting glucose and sorbitol to an acid heat treatment. The production cost of polydextrose is also low, but it is said that bitterness is produced in the process of acidification, and that bitterness affects the flavor of food. (See Patent Document 2 below).

JP 2009-142233 A JP-A-5-255402

Dietary Fiber Fundamentals and Applications Third Edition Daiichi Publishing Co., Ltd. 65 pages 2008

  Therefore, the main object of the present invention is to provide a sugar composition that does not adversely affect the flavor of food and drink.

  In order to solve the above-mentioned problems, intensive studies were conducted on a sugar composition containing dietary fiber, and as shown in the following formula, dietary fiber having a degree of polymerization of 3 to 4 with glucose as a constituent sugar, and glucose as a constituent sugar. If it is a saccharide composition containing dietary fiber having a degree of polymerization of 5 to 9 and a dietary fiber having glucose as a constituent sugar each containing a specific amount of dietary fiber having a degree of polymerization of 10 or more having glucose as a constituent sugar, food and drink The present inventors have found that when used in the present invention, it does not adversely affect the flavor of the food and drink, and can improve taste and quality such as aftertaste.

That is, the present invention relates to the following.
A sugar composition containing dietary fiber having glucose as a constituent sugar,
A sugar composition in which the numerical values of x, y, and z in the following formulas are 0.20 ≦ x ≦ 0.75, 0.25 ≦ y ≦ 0.80, and z ≦ 0.30, respectively.
x: Content of dietary fiber with a degree of polymerization of 3 and 4 with glucose as a constituent sugar (g) / Total content of dietary fiber with glucose as a constituent sugar (g)
y: Content of dietary fiber with a degree of polymerization of 5 to 9 using glucose as a constituent sugar (g) / Total content of dietary fiber containing glucose as a constituent sugar (g)
z: Content of dietary fiber with a degree of polymerization of 10 or more with glucose as a constituent sugar (g) / Total content of dietary fiber with glucose as a constituent sugar (g)

In the sugar composition of the present invention, the total content of dietary fiber containing glucose as a constituent sugar is preferably 5% by weight or more based on the total weight of sugars containing glucose as a constituent sugar.
The sugar composition of the present invention can be produced by allowing a transferase to act on a sugar raw material, and the transferase is selected from the group consisting of α-glucan, α-glucooligosaccharide, and glucose. Aspergillus derived from genus Aspergillus, which acts on any one or more sugar raw materials to produce a sugar having an α1,2 glucoside bond and a sugar having an α1,3 glucoside bond, and capable of enzymatic reaction at a temperature of 65 ° C. or higher. It is preferable to use the transferase.
As the sugar raw material for causing the transferase to act, it is preferable to use a partially decomposed starch obtained by partially hydrolyzing starch with an enzyme other than the transferase or an acid.
As an enzyme other than the transferase, α-amylase is preferably used.

Further, the sugar composition of the present invention is preferably used for food and drink. The sugar composition does not affect the flavor of the food or drink, such as not giving a bitter taste or other miscellaneous taste to the food or drink. It is also possible to improve the taste quality. Therefore, it can utilize widely in food fields, such as food / beverage products, health food / beverage products, and food for specified health uses. The food or drink using the same is preferably, for example, a fermented food or drink. In the case of fermented foods and drinks, it is advantageous in that the carbohydrate of the present invention remains without being decomposed even after the end of fermentation, and exerts its effect.
Further, the present invention provides a method for improving taste quality such as sharpness and aftertaste without adversely affecting the flavor of food by using the sugar composition.

  Since the sugar composition of the present invention does not adversely affect the flavor of food and drink, it can be widely used in the food field and the like.

Electrophoresis photograph (a) of transferase that can be used in the present invention, and deduced amino acid sequence (b) The graph which shows an example of the transferase which can be used for this invention, the relationship between temperature and enzyme activity. The graph which shows an example of the transfer enzyme which can be used for this invention, and the relationship between pH and enzyme activity. Example of transferase that can be used in the present invention, temperature stability graph Example of transferase that can be used in the present invention, pH stability graph An example of a reaction using maltose as a raw material using a transferase, an HPLC chart (a) showing the polymerization degree distribution of the reaction solution, and an HPLC chart (b) showing saccharide structural isomers in the reaction solution 2-3 sugars. Example of reaction using maltose as a raw material using transferase, graph (a) showing change in polymerization degree distribution over time, graph showing change over time in α1,3-linked oligosaccharide, α1,3-linked oligosaccharide, dietary fiber content (B)

Hereinafter, the present invention and terms used in the present invention will be specifically described.
<Sugar composition of the present invention>
The present invention is a sugar composition containing dietary fiber containing glucose as a constituent sugar.
And the saccharide | sugar composition of this invention has the main component of a saccharide | sugar composition as a dietary fiber (x) of polymerization degree 3,4 which uses glucose as a constituent sugar, and the foodstuff of polymerization degree 5-9 which uses glucose as a constituent sugar. If it is a fiber (y), it will not specifically limit. Therefore, x, y, z, which are values indicating the degree of polymerization of dietary fiber contained, are 0.20 ≦ x ≦ 0.75, 0.25 ≦ y ≦ 0.80, and z ≦ 0.30, respectively. It is a sugar composition to contain. That is, the main component of the sugar composition may be x and y, and the z component may be a sugar composition of 0.30 or less.
x: Content of dietary fiber having a degree of polymerization of 3 and 4 with glucose as a constituent sugar (g) / Total content of dietary fiber with glucose as a constituent sugar (g)
y: content of dietary fiber having a polymerization degree of 5 to 9 with glucose as a constituent sugar (g) / total dietary fiber content with glucose as a constituent sugar (g)
z: Content of dietary fiber with a degree of polymerization of 10 or more with glucose as a constituent sugar (g) / Total content of dietary fiber with glucose as a constituent sugar (g)

At this time, preferably 0.20 ≦ x ≦ 0.75, 0.28 ≦ y ≦ 0.75, z ≦ 0.30, more preferably 0.25 ≦ x ≦ 0.75, 0.28 ≦ y ≦ 0.75, z ≦ 0.15, and more preferably 0.20 ≦ x ≦ It is preferable that 0.70, 0.30 ≦ y ≦ 0.75, and z ≦ 0.02.
The sugar composition of the present invention can be produced by the production method of the present invention described later. However, by adjusting, separating, and purifying the enzyme degradation conditions, dietary fibers having a degree of polymerization of 3 to 4 with glucose as a constituent sugar ( x) It is also possible to adjust each content of dietary fiber (y) having a polymerization degree of 5 to 9 having glucose as a constituent sugar and dietary fiber (z) having a polymerization degree of 10 or more having glucose as a constituent sugar.
For example, the content of dietary fiber (z) having a polymerization degree of 10 or more with glucose as a constituent sugar can be further reduced from z ≦ 0.30. In other words, the numerical value range of z can be 0 <z, and z ≦ 0.15, z ≦ 0.02, and z = 0.
Further, for example, the content of dietary fiber (x) having a polymerization degree of 3 or 4 with glucose as a constituent sugar and dietary fiber (y) with a polymerization degree of 5 to 9 with glucose as a constituent sugar, preferably 0.20 ≦ x ≦ 0.75, 0.25 ≦ y ≦ 0.80, more preferably 0.25 ≦ x ≦ 0.75, 0.28 ≦ y ≦ 0.75, and more preferably 0.25 ≦ x ≦ 0.70, 0.30 ≦ y ≦ 0.75. Is advantageous.

That is, in the dietary fiber containing glucose as a constituent sugar, a dietary fiber having a polymerization degree of 3 or 4 using glucose as a constituent sugar (hereinafter also referred to as “dietary fiber having a polymerization degree of 3 or 4”) and glucose are used as constituent sugars. It is desirable to increase the ratio (x, y) of dietary fiber having a polymerization degree of 5 to 9 (hereinafter also referred to as “dietary fiber having a polymerization degree of 5 to 9”) and use it in foods and drinks.
By reducing the proportion (z) of dietary fiber with a degree of polymerization of 10 or more, it becomes possible to make it more easily water-soluble dietary fiber. Both are good.
Therefore, by using dietary fiber with a polymerization degree of 3 or 4 and dietary fiber with a polymerization degree of 5-9 as the main component of dietary fiber with glucose as a constituent sugar, it can be easily dissolved when used in food and drink. It is easy to use for solids and liquids.

<Dietary fiber>
Dietary fiber is a kind of carbohydrate, which is either water-soluble or sparingly water-soluble. Since the sugar composition of the present invention is water-soluble, it is easily dispersible in water and has a higher utility value as an additive for beverages, foods and the like than those with poor water solubility.
The dietary fiber in the present invention is a high-performance liquid chromatographic method described in the enzyme-HPLC method (Eshin No. 13 (Analytical methods for nutritional components in nutrition labeling standards, etc., April 26, 1999)). This method is also used in the AOAC INTERNATIONAL total dietary fiber quantification method (AOAC 2001.03)). In this method, components of three or more sugars remaining after the enzyme treatment are quantified as dietary fiber.
The enzyme-HPLC method is generally used as a method for analyzing dietary fiber. In the present invention, the content of dietary fiber and the degree of polymerization thereof are the content of sugar composition after enzyme treatment and the degree of polymerization in enzyme-HPLC method. And

The HPLC conditions for measuring the dietary fiber content for each degree of polymerization in the enzyme-HPLC method are as follows.
Column: CK04S (Mitsubishi Chemical Corporation),
Column temperature: 65 ° C
Moving bed composition: DW (distilled water)
Detection: RI (differential refraction) detection Flow velocity: 0.35 mL / min

  In the enzyme-HPLC method, when separation of dietary fiber by HPLC is difficult, identification is performed using an oligosaccharide having a known degree of polymerization. In particular, since a dietary fiber having a polymerization degree of 10 is difficult to separate, a component after the elution time of maltooligosaccharide (10 sugars) was defined as a dietary fiber having a polymerization degree of 10.

  The dietary fiber having a polymerization degree of 3 or 4, the dietary fiber having a polymerization degree of 5 to 9, and the dietary fiber having a polymerization degree of 10 or more include α1,2 glucoside bond (α1,2 bond) and / or Alternatively, α1,3 glucoside bond (α1,3 bond) [hereinafter also referred to as “α1,3 bond and / or α1,3 bond carbohydrate”]. Those containing a saccharide having the following formula are preferred.

  The total content of dietary fiber containing glucose as a constituent sugar (preferably a dietary fiber having a polymerization degree of 3 to 9) is preferably 5% by weight or more, more preferably 20%, based on the total weight of carbohydrates containing glucose as a constituent sugar. From the viewpoint of efficacy, it is preferable to set the weight% or more.

<Method for producing sugar composition>
Next, an example of the manufacturing method of the sugar composition of this invention is demonstrated below.
Although the manufacturing method of the saccharide | sugar composition of this invention is not specifically limited, Desirably, it can manufacture by making an enzyme act on saccharide | sugar raw materials, such as a starch decomposition product.

<Description of enzyme>
An example of an enzyme that can be used for production of the sugar composition of the present invention is a transferase, and is a starch degradation product (including a partial starch degradation product) containing at least one selected from α-glucan, α-glucooligosaccharide, and glucose. It acts on sugar raw materials such as to produce the dietary fiber of the present invention, and has the property of producing a sugar composition characterized by the degree of polymerization of the dietary fiber.
More preferably, the transferase has the property of acting on a sugar raw material to produce a sugar having an α1,2 glucoside bond (α1,2 bond), an α1,3 glucoside bond (α1,3 bond), An enzyme reaction is possible at a temperature of 65 ° C. or higher, and Aspergillus transferase can be exemplified as the thermostable enzyme. Hereinafter, the enzymes exemplified here will be described.

A saccharide containing an α1,2 bond means a saccharide having two or more saccharides containing at least one α1,2 bond in the molecule. In addition to an oligosaccharide consisting of only an α1,2 bond, Oligosaccharides consisting of other bonds are also included.
Specifically, cordobiose [O-α-D-glucopyranosyl- (1 → 2) -OD-glucopyranose]], which is a disaccharide, and cordobiosyl glucose [O-α-D-glucopyranosyl, which is a trisaccharide, -(1 → 2) -O-α-D-glucopyranosyl- (1 → 4) -D-glucopyranose] and the like.
In addition, it is possible to determine whether 1,2 bonds are contained in the structure of trisaccharide or higher components by methylation analysis (Journal of Biochemistry Vol. 55, page 205, 1964) or NMR analysis.

The carbohydrate containing α1,3 bond means that the molecule contains one or more α1,3 bonds, in addition to oligosaccharides consisting of only α1,3 bonds, α1,3 bonds and other bonds. An oligosaccharide consisting of Specifically, in addition to nigerose [O-α-D-glucopyranosyl- (1 → 3) -OD-glucopyranose], which is a disaccharide, nigerosyl glucose [O-α-D-glucopyranosyl, which is a trisaccharide. -(1 → 3) -O-α-D-glucopyranosyl- (1 → 4) -D-glucopyranose], nigerotriose [O-α-D-glucopyranosyl- (1 → 3) -O-α-D -Glucopyranosyl- (1 → 3) -D-glucopyranose] and the like.
Moreover, it can be judged by methylation analysis method, NMR analysis method, etc. also about the component more than 3 sugars whether the structure contains a 1,3 bond.

The carbohydrate containing α1,2 bond and α1,3 bond means that the molecule contains one or more α1,2 bond and α1,3 bond, respectively, and α1,2 bond and α1,3 bond. And oligosaccharides composed of other bonds.
Here, when there are more α1,2 bonds than α1,3 bonds in the molecule, the saccharide includes α1,2 bonds, and when there are more α1,3 bonds than α1,3 bonds, α1 , Carbohydrate containing 3 bonds.

  The determination of whether a 1,2-bond or a 1,3-bond is contained in the structure of a component having three or more sugars can also be determined by the dietary fiber content calculated by the enzyme-HPLC method.

  By the way, the binding mode between glucose in starch sugar is α1,4 bond and α1,6 bond, and normal starch sugar has almost no α1,2 bond or α1,3 bond. From the analysis example using the enzyme-HPLC method described above, when the starch is acid roasted, the content of 1,2 bonds and / or 1,3 bonds (α and β binding modes are unknown) increases and dietary fiber It is described that the content increases (Journal of Applied Glycoscience 49, 479, 2002). However, when the transferase of the present invention is used, α1,3 and α1,3 linked carbohydrates are markedly increased. Can be increased.

The transferase has a residual activity of 90% or more at 65 ° C. and acts on a sugar solution containing at least one selected from α-glucan, maltooligosaccharide, and glucose, and a carbohydrate containing α1,2 bond and α1,3 bond An enzyme that exhibits the action of producing
Filamentous fungi (Absidia, Acremonium, Actinomadura, Alternaria, Aspergillus, Chaetomium, Coprinus, Coriolus, Geotrichum, Humicola, Monascus, Mortierella, Mucor, Nocardiopsis, Oidiodendron, Penicillium, Rhizomucor, Rhizopus, Trichoderma, Verticillium)
Basidiomycetes (Coliolus, Corticium, Cyathus, Irpex, Polyporus, Pynoporus, Trametes),
Bacteria (Aeromonas, Agrobacterium, Alcaligenes, Agrobacterium, Alteromonas, Arthrobacter, Bacillus, Brevibacterium, Chromobacterium, Corynebacterium, Crypnohectria, Erwinia, Escherichia, Flavobacterium, Klebsiella, Lactobacillus, Lactococcus, Leuconostoc, Microbacterium, Micrococcus, Pimelobacter, Plesiomonas, Protaminobacter, Pseudomonas, Serratia, Streptococcus , Streptoverticillium, Sulfolobus, Thermus, Xanthomonas)
Actinomydra (Actinomadra, Actinomyces, Actinoplanes, Amycolatopsis, Eupenicillium, Nocardiopsis, Streptomyces, Thermomonospora)
A strain having an example of use in food production such as yeast (Aureobasidium, Candida, Irpex, Kluyveromyces, Pycnoporus, Saccharomyces, Trichosporon) is desirable.

Among them, Aspergillus genus such as Aspergillus niger genus or Aspergillus awamori genus is preferable, among them Aspergillus niger (ATCC [American Type Culture Collection] 10254), Aspergillus niger van Tiger. niger fsp. Henbergii Blochwitz ex Al-Musallam (NBRC [NITE Biological Resource Center] 4043), Aspergillus awamori (ATCC 14331), etc. are mentioned as suitable strains.
When obtaining a target transferase using a bacterium belonging to the genus Aspergillus, a known method is appropriately employed for the culture. For example, both liquid culture and solid culture can be arbitrarily used. .
Microorganisms to be used are not limited to wild strains. For the above wild strains, ultraviolet rays, X-rays, radiation, various chemicals [NTG (N-methyl-N′-nitro-N-nitrosoguanidine), EMS (ethyl methanesulfonate), etc.], etc. Mutant strains mutated by the artificial mutation means used can also be used as long as they are thermostable transferases that produce carbohydrates having α1,2 bonds and α1,3 bonds.

  As the carbon source of the medium used in the culture using Aspergillus bacteria, for example, glucose, fructose, sucrose, lactose, starch, glycerin, dextrin, lecithin and the like are used alone or in combination, and nitrogen As the source, both organic and inorganic nitrogen sources can be used. Among them, examples of the organic nitrogen source include peptone, yeast extract, soybean, kinako, rice bran, corn steep liquor, meat extract, casein, amino acid. On the other hand, ammonium sulfate, ammonium nitrate, phosphate-ammonium phosphate, diammonium phosphate, ammonium chloride, etc. will be used as the inorganic nitrogen source. Furthermore, examples of inorganic salts and micronutrients added to such a medium include sodium, magnesium, potassium, iron, zinc, calcium, manganese salts, vitamins, and the like. Moreover, it is also possible to use natural products, such as wheat bran, as a culture medium component containing said various components.

  Cultivation using Aspergillus bacteria is generally performed at a temperature of 10 to 40 ° C., preferably a culture temperature of 25 to 30 ° C. is advantageously employed, and the medium pH is 2.5 to 8.0. I just need it. The required culture period varies depending on the cell concentration, medium pH, medium temperature, medium composition, etc., but is usually about 4 to 9 days, and when the target transferase has reached its maximum. The culture is stopped.

In this way, after culturing the microorganism, the transferase is recovered. The activity of the transferase is found in both the cells of the culture and the medium, and can be purified and used by a known method.
As an example, after dialysis of a crude enzyme preparation obtained by concentrating a treated product of a culture solution, anion exchange column chromatography using a gel “TOYOPEARL DEAE-650M” manufactured by Tosoh Corporation, followed by GE Healthcare Gel filtration chromatography in which the columns “HiLoad 16/60 upperdex 200 pg” and “HiLoad 16/60 Superdex 75 pg” manufactured by Japan are combined, and then the column “MonoP 5/200 GL” manufactured by GE Healthcare Japan is used. By performing the isoelectric point chromatography used, a single enzyme can be obtained electrophoretically.

Enzymatic chemistry when Aspergillus-derived enzyme is used is as follows.
(1) Action An α-glucosidase that cleaves the α-glucoside bond at the non-reducing end of the substrate into an exo-type, which hydrolyzes α1,2 and α1,3 bonds in addition to α1,4 bonds, while sugar donors Glucose transfer reaction is also performed in which the glucose residue from is transferred to the hydroxyl group at any of the 2, 3, and 4 positions of the glucose residue of the sugar receptor.
Since the transfer product in which the α-glucosyl group is transferred to the hydroxyl group at the 4-position is decomposed again by the present enzyme, a saccharide having α1,2, α1,3 bonds is finally accumulated as the transfer product. In addition, even when a derivative having a glucose skeleton or a compound having an OH group is used as a receptor, it has transglycosylation activity. Accordingly, glycosides can be collected by acting on a substrate obtained by adding a glucose derivative to α-glucan, α-glucooligosaccharide, glucose and the like.
As the substrate, α-glucan such as starch, amylose, amylopectin, glycogen, dextrin, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose, maltooligosaccharide, cordobiose, α-glucooligosaccharide such as nigerose, Glucose can be used.

(2) Molecular weight Based on native-gel electrophoresis, the molecular weight is 110,000 to 90,000 daltons, preferably 97,000 daltons. At this time, Native-gel electrophoresis analysis may be performed by GE Healthcare Japan Co., Ltd., and FastSystem.
From SDS-gel electrophoresis, two bands having a molecular weight of 60,000 to 40,000 daltons, preferably a molecular weight of 48,000, and a molecular weight of 70,000 to 50,000 daltons, preferably a molecular weight of 59,000 daltons were obtained. Have. At this time, the SDS-gel electrophoretic analysis may be performed by PhasSystem, manufactured by GE Healthcare Japan.
(3) Isoelectric point The isoelectric point of the enzyme is pI: 4.0 to 6.0, preferably 4.9 to 5.5, and the center value is 5.2. At this time, isoelectric focusing (Phast System, manufactured by GE Healthcare Japan) may be performed together with a standard protein having a known isoelectric point.
(4) Optimum temperature It is 60-70 degreeC by the reaction for pH 4.0 and 10 minutes, Preferably it is 65 degreeC.
(5) Optimum pH
After reaction at 50 ° C. for 10 minutes, the pH is 3.0 to 4.0, preferably pH 3.5.
(6) Temperature stability (heat resistance)
After treatment at 65 ° C. for 30 minutes, 80% or more, preferably 90% or more, more preferably 95% or more of the initial activity remains.
(7) pH stability When stored at 4 ° C. for 24 hours, the pH is 2.5 to 5.5, preferably pH 3.0 to 5.0.

(8) Substrate specificity At least one of nigerose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose, particularly high affinity for nigerose or high reactivity Those having the following are preferred. In addition, maltose (each 10 mM solution) may be used as a reference.
(9) Influence of metal ions Those which are inhibited by zinc ions and copper ions are preferred. Those that are activated with magnesium ions and manganese ions to the same level or higher are preferred.
(10) Amino acid sequence Those having one or more of the following amino acid sequences are preferred.
Sequence number 1: LLVEYQTDERLHVMIYDADEVYQVPESVLPR
Sequence number 2: TWLPDDPYVYGLGEHSDPMR
Sequence number 3: IPLETMWTDIDYMDKR
SEQ ID NO: 4: VFTLDPQR
Sequence number 5: WASLGAFYTFYR

  In the amino acid sequence shown in each SEQ ID NO, the amino acid sequence in which one or several amino acids are substituted, deleted or added means an amino acid sequence functionally equivalent to each SEQ ID NO. An amino acid sequence in which 1 to 6, preferably 1 to 6, more preferably 1 to 3 amino acids are substituted, deleted or added, and still has the enzymatic property. In addition, addition includes addition of 1 or several, preferably 1 to 6, more preferably 1 to 3 amino acids to both ends.

<Specific example of sugar composition production method>
The transferase (preferably the transferase obtained from the above-mentioned cells and culture medium) is allowed to act on a sugar raw material (substrate) comprising at least one selected from α-glucan, α-glucooligosaccharide, and glucose as substrates. Thus, a sugar composition containing a saccharide having α1,2 and / or α1,3 bonds can be obtained.
Examples of substrates (sugar raw materials) that can efficiently produce carbohydrates containing α1,2 bonds and α1,3 bonds include α-glucans such as starch, amylose, amylopectin, glycogen, dextrin, maltose, maltotri It is possible to use α-glucooligosaccharide such as oose, maltotetraose, maltopentaose, maltohexaose, malto-oligosaccharide, cordobiose, nigerose, and glucose alone or in combination.

Moreover, you may use the starch degradation product obtained by partially hydrolyzing starch substances, such as starch, amylopectin, and amylose, with an amylase or an acid as a substrate (sugar raw material). “Partially hydrolyzing” includes, for example, adjusting to a specific DE (dextrose equivalent).
At this time, the starch (partial) decomposition product (preferably starch liquefaction liquid) is DE (dextrose equivalent) 5-40, preferably DE8-30. Moreover, although starch is not specifically limited, potato starch, wheat starch, corn starch, waxy corn starch, etc. are mentioned, You may use these in combination of 1 type, or 2 or more types.
Examples of amylases that partially hydrolyze starch include α-amylase, maltopentaose-producing amylase, maltohexaose described in Handbook of Amylases and Related Enzymes (Pergamon Press, Tokyo, 1988). A generation amylase or the like is used. A combination of these amylases and debranching enzymes such as pullulanase and isoamylase can also be advantageously carried out.

When glucose is used as a substrate (sugar raw material), unlike other substrates, carbohydrates containing α1,2 and α1,3 bonds are produced by a condensation reaction that is the reverse reaction of the hydrolysis reaction. However, since the yield is lower than that of the transfer reaction, carbohydrates higher than maltose are desirable from the economical point of view.
As an α-glucosyl group receptor, it is only necessary to have a glucose residue at the non-reducing end. Specifically, glucose, maltose, maltotriose, cordierbiose, cordierioose, cordierbiosylglucose, nigerose, Nigerotriose, nigerosyl glucose, isomaltose, isomaltotriose, panose, cellobiose, sophorose, laminaribiose, gentiobiose, trehalose, sucrose and the like. A glucose derivative or a compound having an OH group can also serve as a receptor.

When an enzyme that generates α1,2 bond or α1,3 bond is allowed to act on a substrate (sugar raw material), it is hydrolyzed with α-amylase, β-amylase, glucoamylase, α-glucosidase, etc., or branching enzyme It is also possible to adjust the molecular weight, sweetness, reducing power, etc., or reduce the viscosity by acting glucosyltransferase, etc.
Among these enzymes, if a thermostable enzyme that does not inactivate at 50 ° C. to 65 ° C. is selected, it can be treated at a high temperature exceeding 50 ° C. when acting together with the transferase of the present invention.
These enzymes may be allowed to act simultaneously with transferases that generate α1,2 bonds and α1,3 bonds, or may be reacted separately. When the other enzyme is a starch degrading enzyme, when it is caused to act simultaneously with the transferase of the present invention, the production of a partially decomposed starch and the production of sugar from the decomposed product proceed simultaneously. In addition, it is optional to carry out further processing such as hydrogenation of the obtained saccharides having α1,2 bonds and α1,3 bonds to form sugar alcohols to eliminate the reducing power.

The substrate concentration in the reaction may be a concentration that dissolves in the reaction solution, in the range of 1 to 80% by weight, in order to obtain a carbohydrate more efficiently, 10 to 60% by weight, more preferably 20 to 45% by weight. It is desirable to carry out under the following conditions.
The temperature used for the reaction may be a temperature range in which the enzyme is stable in the reaction solution, and it is appropriate to perform the reaction at 50 to 90 ° C, preferably 60 to 80 ° C, more preferably 60 to 70 ° C. The pH used for the reaction is usually from 3.5 to 7.0, preferably from 4.0 to 6.5, more preferably from 4.0 to 6.0. When a plurality of enzymes are used for the reaction, the above reaction temperature and reaction pH may be appropriately adjusted according to the enzyme used and the enzyme reaction step.
The reaction period is 10 minutes to 7 days, preferably 1 hour to 4 days.
A higher enzyme concentration used for the reaction is advantageous because it shortens the reaction time. If the enzyme concentration is low, the yield of saccharides having α1,2 bonds and α1,3 bonds will deteriorate.

Reaction solutions containing carbohydrates with α1,2 bonds and α1,3 bonds are filtered, centrifuged, etc. to remove insoluble matters, then decolorized with activated carbon, H-type, OH-type ion exchange Desalinate with resin and concentrate to syrup product. Furthermore, it is optional to dry it into a powdered product.
If necessary, further advanced purification (separation purification) is optional. Thereby, the dietary fiber content in the sugar composition can be adjusted, and the dietary fiber content in which glucose is a constituent sugar can be adjusted with respect to the total weight of carbohydrates in which glucose is a constituent sugar.
For example, it can be highly purified by fractionation by ion exchange (cation exchange and anion exchange) column chromatography, fractionation by activated carbon column chromatography, or fractionation by gel filtration column chromatography.

As ion exchange column chromatography, column chromatography using a cation exchange resin using a salt-type strongly acidic cation exchange resin disclosed in JP-A-58-23799, JP-A-58-72598, and the like, A method of removing contaminating saccharides and collecting a high-content fraction can be advantageously carried out. It is desirable to adjust the dietary fiber content and the like in the sugar composition by a column chromatography method using a cation exchange resin using a salt-type strongly acidic cation exchange resin.
At this time, it is optional to adopt any of a fixed floor method, a moving floor method, and a simulated moving floor method.

  Although the more detailed specific example of the saccharide | sugar composition of this invention is demonstrated below, this invention is not limited to this.

<Example 1>
30% by weight of DE12 potato starch liquefied liquid (starch partially decomposed product) was adjusted to a temperature of 65 ° C. and a pH of 6.0, and then the thermostable transferase purified by the method of Reference Examples 1 and 2 described later was used with a solid content of 3 U. α-amylase (Termamyl 120L, manufactured by Novozymes) was added at 0.005% by weight of solid content and allowed to act for 36 hours. The starch decomposition product solution was subjected to activated carbon / ion purification treatment / concentration to obtain 75% by weight of the sugar composition of Example 1.
DE is dextrose equivalence (DE), and is an indicator of the degree of degradation of a starch degradation product. Smaller values are larger in molecule and higher in viscosity. The titer of thermostable transferase and the measurement method will be described later.

<Example 2>
After adjusting 35% by weight of DE18 corn starch liquefied liquid (starch partially decomposed product) to a temperature of 50 ° C. and a pH of 5.5, the heat-resistant transferase purified by the method of Reference Example 1 described later acts on a solid content of 30 U for 24 hours. I let you. Α-amylase (Termamyl 120L, manufactured by Novozymes) vs. solid content of 0.15% by weight, and glucoamylase (AMG, manufactured by Novozymes) vs. solid content of 0.1% by weight are added to the sugar solution after the reaction to act for 24 hours. I let you. This starch decomposition product solution is activated carbon, ion-purified, concentrated, prepared into a 50 wt% aqueous solution, and packed with a strongly acidic cation exchange resin (FX1040, manufactured by Organo) heated to 60 ° C. Using an apparatus (Tresone, manufactured by Organo), low molecules were removed.
The fractionated starch degradation product solution was subjected to activated carbon / ion purification, then concentrated, and powdered with a spray dryer to obtain the carbohydrate of Example 2.

<Example 3>
After adjusting 40% by weight of DE10 waxy corn starch liquefied liquid (starch partially decomposed product) to a temperature of 65 ° C. and a pH of 5.5, the thermostable transferase purified by the method of Reference Examples 1 and 2 described later is used with a solid content of 10 U, α-Amylase (Termamyl 200L, Novozymes) was added at 0.05% by weight of solid content and allowed to act for 48 hours. To the sugar solution after the reaction, glucoamylase (AMG, manufactured by Novozymes) vs. 0.3 wt% solid content was added and allowed to act at 40 ° C. for 24 hours. This starch decomposition product solution is activated carbon / ion-purified / concentrated, prepared to a 50% by weight aqueous solution, and subjected to a gel filtration column (TOYOPEARL HW-40S, manufactured by Tosoh Corporation) heated to 60 ° C. Removed.
The fractionated starch degradation product solution was subjected to activated carbon / ion purification, concentrated, and powdered with a spray dryer to obtain the carbohydrate of Example 3.

<Example 4>
After adjusting 20% by weight of DE8 corn starch liquefied liquid (starch partially decomposed product) to a temperature of 60 ° C. and a pH of 6.0, the heat-resistant transferase purified by the method of Reference Example 1 described later was added to a solid content of 6 U, α-amylase ( Termamyl 120L (manufactured by Novozymes) with a solid content of 0.02% and β-amylase (β-amylase # 1500, manufactured by Nagase Seikagaku Corporation) with a solid content of 0.03% by weight were added and allowed to act for 36 hours. The starch decomposition product solution was subjected to activated carbon / ion purification treatment, concentrated, and powdered with a spray dryer to obtain the saccharide of Example 4.
In addition, the starch liquefaction liquid of Examples 1-4 can be prepared by the enzyme liquefaction by (alpha) amylase (Termamyl 120L, Novozymes company make). Moreover, what was concentrated can be used for the thermostable transferase used in Examples 1-4. The addition amount of the thermostable transferase used in Examples 1 to 4 indicates the enzyme amount (U) per gram of the solid content.

<Example 5>
The sugar composition of Example 4 and maltose (special grade) manufactured by Kanto Chemical Co., Ltd. were mixed at a ratio of 18:82 per solid content and dissolved by adding water. Powdered to obtain the sugar composition of Example 5.

<Example 6>
20% by weight of DE30 corn starch liquefied liquid (starch partially decomposed product) prepared by acid liquefaction with hydrochloric acid was adjusted to a temperature of 65 ° C. and pH 6.5, then purified by the method of Reference Example 1 described later, and concentrated heat resistance. 0.5 U per gram of transferase, α-amylase (Termamyl 200L, Novozymes) vs. 0.1% solids, debranching enzyme (Chrytase PLF, Amano Enzyme) vs. 0.3% solids Added and allowed to act for 96 hours. The starch decomposition product solution was subjected to activated carbon / ion purification, concentrated, and powdered with a spray dryer to obtain the saccharide of Example 6.

<Example 7>
After adjusting 45% by weight of DE30 potato starch liquefied liquid (starch partially decomposed product) prepared by enzyme liquefaction with α-amylase (Termamyl 120L, manufactured by Novozymes) to a temperature of 70 ° C. and pH 4.0, Reference Example 1 to be described later After purification by the method 2, the concentrated thermostable transferase is 60 U per gram of solid content, α-amylase (Termamyl 200L, Novozymes) is 0.5% by weight of solid content, debranching enzyme (Chrytase PLF, Amano) Enzyme)) 0.8% by weight of the solid content was added and allowed to act for 1 hour. To the sugar solution after the reaction, glucoamylase (AMG, manufactured by Novozymes) vs. 0.3 wt% solid content was added and allowed to act at 40 ° C. for 24 hours. This starch degradation product solution is activated carbon, ion purified and concentrated, prepared to a 50 wt% aqueous solution, and subjected to a gel filtration column (TOYOPEARL HW-40S, manufactured by Tosoh Corporation) heated to 60 ° C. Removed.
The fractionated starch degradation product solution was subjected to activated carbon / ion purification, concentrated, and powdered with a spray dryer to obtain the carbohydrate of Example 7.

<Comparative Examples 1-3>
As Comparative Example 1, trade name “Fibersol 2” (digestible dextrin) manufactured by Matsutani Chemical Industry Co., Ltd., as trade name “IMO900P” manufactured by Showa Sangyo Co., Ltd. as Comparative Example 2, Kanto as Comparative Example 3 Chemical maltose (special grade) was used.

<Sugar composition>
About the saccharide | sugar composition of Examples 1-7 and Comparative Examples 1-3, it comprises the dietary fiber of polymerization degree 3 and 4 which uses glucose as a constituent sugar, the dietary fiber of 5 to 9 degree of polymerization which uses glucose as a constituent sugar, and glucose. The total of dietary fiber having a polymerization degree of 10 or more as sugar and the dietary fiber having glucose as a constituent sugar is determined by an enzyme-HPLC method. From the obtained content, x, y, z, ie, the total of dietary fiber, The ratio (weight ratio) of dietary fiber for each degree of polymerization was determined.

As is apparent from Table 1 above, the sugar compositions of Examples 1 to 7 have at least the same total content of dietary fiber as measured by the enzyme-HPLC method as compared to commercially available products (Comparative Examples 1 to 3). There may be more than a commercial thing.
When a dietary fiber material is added to a food or drink, the effect is sufficiently exhibited if the dietary fiber content of glucose as a constituent sugar is 5% by weight or more with respect to the total weight of carbohydrates as a constituent sugar of glucose. Moreover, the saccharide | sugar composition of Examples 1-5 is soluble in water, and it turns out that the saccharide | sugar composition by this invention has the characteristic as water-soluble dietary fiber.
In the sugar compositions of Examples 1 to 7 produced by the thermostable enzyme, the values of x, y, and z are in the ranges of 0.20 ≦ x ≦ 0.75, 0.25 ≦ y ≦ 0.80, and z ≦ 0.30, respectively. On the other hand, Comparative Examples 1 to 3 and the commercially available sugar composition are at least one of x, y, and z outside the scope of the present application. Compared with known products, the sugar composition of the present invention is a food. It can be seen that it clearly exhibits different properties in terms of fiber composition.
In addition, as will be described later, Examples 6 and 7 are considered to be the best in terms of flavor, sharpness, and aftertaste in a liquid, semi-solid, and solid food or drink with a good balance. Moreover, about z, it is thought that it is hard to influence the flavor of food-drinks, so that there is little as a tendency.
Therefore, sugar compositions in which the numerical values of x, y, and z are in the ranges of 0.20 ≦ x ≦ 0.75, 0.25 ≦ y ≦ 0.80, and z ≦ 0.30 are preferable, and z may be 0.

Next, the following evaluation test (food addition test) was performed on the sugar compositions of Examples 1 to 7 and Comparative Examples 1 to 3. In the evaluation test, 9 subjects have sampled the sugar composition-added food, and when 8 or more responded that they were good, ◎, when 7-6 answered that they were good, ○, 5-4 A case where a person answered that the person was good was evaluated as △, and a case where three or less persons answered that it was good was evaluated as x.
Flavor is good: Does not adversely affect the original flavor of food (not affected by bitterness, bitterness, smell, etc.).
Good sharpness: The taste is refreshing (the taste that tastes in the mouth is less).
Good aftertaste: Good taste quality in the mouth after eating and drinking.
In addition, the dietary fiber content in the following Tables 2 to 13 is the amount of dietary fiber in the food produced in each test.
<Food additive test 1>
An evaluation test was performed on a 5% by weight aqueous solution of each sugar composition. The results are shown in Table 2 below.

<Food additive test 2>
Wort extract 40g, maltose 100g, hop extract 5g, soybean peptide 1.7g, yeast 0.6g in Examples 1, 2, 6 or Comparative Examples 1, 2, 3 (prepared to 75% solid content) Was mixed to 30 g and adjusted to a final weight of 1000 g by adding water, followed by fermentation for 7 days. Regarding Example 5, 125 g of Example 5 was mixed with 40 g of wort extract, 5 g of hop extract, 1.7 g of soybean peptide, and 0.6 g of yeast, and after adding water to adjust the final weight to 1000 g, 7 days Fermentation was performed. After fermentation, the yeast was removed with diatomaceous earth to produce a beer (sparkling liquor). The evaluation test is shown in Table 3 below.

<Food additive test 3>
350 g of wheat flour, 15 g of sugar, 30 g of dry yeast, 1.5 g of yeast food, 10 g of sodium chloride, 15 g of skim milk powder and 200 g of water were mixed to produce a medium seed. The sugar compositions of the powder Examples and Comparative Examples are preliminarily added with water to form a suspension having a solid content of 75%. After adding 33 g of the sugar composition of Example or Comparative Example in terms of solid content, the mixture was kneaded with a mixer for 15 minutes. Next, the mixed dough was divided and rolled to produce an intermediate dough. Next, the intermediate dough was put in a polyethylene bag, rapidly frozen, and stored in a freezer at -30 ° C. for one week. After freezing for one week, it was thawed and fermented using a dough conditioner. Then, the fermented dough was divided, re-fermented with a proofer, then baked to make a prototype of bread. The evaluation results are shown in Table 4 below.

<Food additive test 4>
110 g of unsalted butter softened with a microwave oven was kneaded with a hand mixer until it became creamy, and then the sugar composition of the example or the comparative example was added in a total of 100 g in three portions while mixing. Next, 100 μg of vanilla essence was added, and 29 g of egg yolk was added in three portions and kneaded. Then, 124 g of weak flour and 2.5 g of baking powder were added and lightly mixed, and then rolled into a 3 cm diameter ball. Next, they were placed on oven paper, lightly crushed to a uniform thickness, and then baked in an oven preheated to 160 ° C. for about 30 minutes to produce a cookie. The evaluation results are shown in Table 5 below.

<Food additive test 5>
1 g of sugar was added to 97 g of black tea extract, and 2 g of the sugar composition of the example or comparative example was added to make a black tea. The evaluation results are shown in Table 6 below.

<Food additive test 6>
36g of Examples or Comparative Examples (prepared to a solid content of 75% in advance) were added to 38g of fermented skim milk, 13g of stabilizer, 0.35g of fragrance, and 0.05g of water, respectively, and homogenized with a homogenizer to drink yogurt Prototyped. The evaluation results are shown in Table 7 below.

<Food additive test 7>
25 g of shochu shellfish (triangle, manufactured by Sapporo Beer) with an alcohol content of 20% by weight, 2 g of the sugar composition of the example or comparative example were added, water was added to a total weight of 100 g, and the mixture was stored at 4 ° C. for 1 day. An alcoholic beverage was prototyped. The evaluation results are shown in Table 8 below.

<Food additive test 8>
Salt 0.5g, vitamin C 0.03g, vitamin B1 0.03g, magnesium chloride 0.2g, calcium lactate 0.2g, citric acid 2.4g, sodium citrate 1.7g, flavor 2g, glucose 80g, confectionery 13g Then, 60 g of the sugar composition of Example or Comparative Example was mixed with 1500 g of water, and heat sterilized to produce a sports drink as a prototype. The evaluation results are shown in Table 9 below.

<Food additive test 9>
Add and mix 4 g of the sugar composition of the example or comparative example (prepared to a solid content of 75% by weight) into 4.5 g of unsalted butter, 10.5 g of skim milk powder, 11 g of sugar, 8 g of egg yolk, and 62 g of water. Ice cream was produced. The evaluation results are shown in Table 10 below.

<Food additive test 10>
26.7 g of dried egg yolk reconstituted with 51.3 g of water, 36.0 g of sugar, and 36 g of the sugar composition of the example or comparative example were put into a bowl and mixed with a frothing device. 16.0 g of sieved flour was added and further mixed with a frothing device. To this, 200 g of milk warmed to 50 ° C. was added little by little, and after passing through a strainer, it was stirred until it became creamy over medium heat to produce a custard cream. The evaluation results are shown in Table 11 below.

<Food additive test 11>
A uniform slurry was prepared by adding the same amount of water to 100 g of raw bean paste. Next, 100 g of the sugar composition of Examples / Comparative Examples in terms of solid content was added to the slurry and boiled to prepare a red bean kojian prototype. The evaluation results are shown in Table 12 below.

<Food additive test 12>
30 g of Examples or Comparative Examples (prepared to a solid content of 75%) were added to 33 g of vinegar, 2 g of sugar, 0.5 g of salt, 8.4 g of thin soy sauce, and 26.3 g of soup stock to make a non-oil dressing. The evaluation results are shown in Table 13 below.

As apparent from Tables 2 to 13 above, the sugar compositions of Examples 1 to 7 are superior to the sugar compositions of Comparative Examples 1 to 3 in all evaluation results such as flavor, sharpness, and aftertaste.
Therefore, sugar compositions with x, y, and z values in the ranges of 0.20 ≦ x ≦ 0.75, 0.25 ≦ y ≦ 0.80, and z ≦ 0.30, respectively, have a dietary fiber content equal to or higher than that of commercial dietary fiber. Nevertheless, it has been demonstrated that it can be used without adversely affecting the flavor of the food in the use of various foods.

  Detailed examples of the transferase used in the production of the sugar composition of the present invention will be described below, but the present invention is not limited thereto.

<Analysis method>
The activity of transferase was evaluated by hydrolysis activity using maltose as a substrate. As the partial activity of the transferase, the hydrolysis activity using maltose as a substrate was measured as follows. To 100 μL of 20 mM maltose, 16 μL of 200 mM acetate buffer (pH 4.0) and 84 μL of enzyme solution were added and reacted at 50 ° C. for 10 minutes. Then, the reaction was stopped by treating in a boiling bath for 5 minutes. The amount of glucose produced in the reaction solution was measured with Glucose CII Test Wako (manufactured by Wako Pure Chemical Industries). The amount of enzyme that decomposes 1 μmol of maltose per minute was defined as 1 U.

The composition analysis of the reaction solution in which various substrates were allowed to act on transferase was carried out using the following analysis method.
The polymerization degree distribution of the reaction solution was analyzed by gel filtration column under the following conditions.
HPLC measurement conditions: Column: Mitsubishi Chemical CK04S,
Column temperature: 65 ° C
Moving bed composition: D.D. W. (Distilled water)
Detection: RI (differential refraction detector)
Flow rate: 0.35 mL / min

  Among the 2, 3 sugar components of the resulting composition, oligosaccharides having α1,2 bonds (Cojibies, Kojibiosylglucose), oligosaccharides having α1,3 bonds (Nigellose, Nigerosylglucose, Nigerotriose) are Analysis was carried out by the phosphate-phenylhydrazine method for labeling the reducing end of sugar (Japanese Patent No. 2846059). Each peak was identified by comparing with the elution time of a standard sugar analyzed in advance under the same conditions, and the production amount was calculated from the peak area.

Commercially available reagents were used as standard sugars for disaccharides, and commercially available reagents and compositions obtained by the following methods were used as standard sugars for trisaccharides. Nigerotriose and nigerosyl glucose were prepared according to the procedure described in Biochimica et Biophysica Acta, Vol. 1700, page 189, 2004, and used as preparations. Codybiosyl glucose and nigerosyl glucose were prepared according to the method described in JP-A No. 2003-169665 and used as preparations.
The specific analysis method was performed as follows.
HPLC measurement conditions: Column: Unison UK-Amino 250 × 4.6 mm (manufactured by Intact Corporation)
Column temperature: 35 ° C

Detector: Fluorescence detector Ex 330nm, Em 470nm
Flow rate: Eluent: 1.0 mL / min, Reaction solution: 0.4 mL / min In order to evaluate whether or not 1,2-bonded and 1,3-bonded saccharides are contained in the whole composition obtained, A methylation analysis method (Journal of Biochemistry, Vol. 55, p. 205, 1964) was performed.
In order to evaluate whether or not 1,2-bonded and 1,3-linked saccharides are contained in the components of 3 or more sugars of the obtained composition, an enzyme-HPLC method was performed. Ingredients containing 3 or more sugars detected by this analysis method were defined as dietary fiber content.
Journal of Applied Glycoscience Vol. 49, page 479 It is known from the analysis example in 2002 that the presence of 1,2 and 1,3 bonds in the molecule of α-glucan increases the dietary fiber content of carbohydrates. Therefore, it is considered that 1,2-bond and 1,3-bond exist in components of 3 or more sugars detected by this analysis method.

<Relationship between low DE component aging and reaction temperature>
Corn starch was liquefied using α-amylase by a conventional method to obtain a starch liquefaction solution having a concentration of 30% and DE10. Subsequently, this starch liquefaction liquid was stored at each temperature of 50, 55, 60, 65, and 70 ° C., and the reaction was continued to decompose to DE28. In addition, DE was measured by the method described in the powdered sugar-related industrial analysis method and the Food Chemical Newspaper Co., Ltd.
Hydrochloric acid was added at the end of the reaction, and the reaction solution was adjusted to pH 4.0 and stored at 80 ° C. for 1 hour to inactivate α-amylase. In order to remove foreign substances in each of the obtained reaction liquids, a filter cloth is put on a 9 cm Nutsche, and 20 g of a filter aid (Celite KC580, manufactured by Celite, USA) is overlaid to form a cake and let the reaction liquid pass. It was. Turbidity (spectrophotometer UV-1200, Shimadzu Corporation, 1 cm quartz cell) was measured in order to evaluate the decomposability and properties of the obtained sample. The results are listed in Table 17 below.

As shown in Table 17, there is no significant change in turbidity at a reaction temperature of 65 ° C. or higher.
A change in turbidity was confirmed at 60 ° C., and a significant increase in turbidity was observed in reactions below 55 ° C. As the reaction temperature decreased, it was considered that the turbidity was increased because the components having high molecular weight and high crystallinity in the starch liquefaction liquid were precipitated and the enzyme could not act. From the above, it can be seen that an enzyme reaction with a low DE component (particularly DE 0 to 20) is preferably performed at a reaction temperature of 65 ° C. or higher.

<Reference Example 1: Production of transferase from ATCC 10254 strain>
As media, starch: 2%, peptone: 0.25%, yeast extract: 0.25%, soybean flour: 1%, monopotassium phosphate: 0.03%, magnesium sulfate: 0.01%, calcium chloride: 0.01% and sodium chloride: 0.01% containing pH = 6.5 was prepared, and 100 mL of the solution was placed in a 500 mL Erlenmeyer flask and steam sterilized, and then the Aspergillus niger ATCC 10254 The strain was inoculated and cultured with shaking at a temperature of 30 ° C. for 3 days.
The above seed was diluted 80-fold with sterilized water, 10 mL of the seed was transplanted into 10 g of wheat bran sterilized by autoclaving contained in a 500 mL Erlenmeyer flask, and stirred well so that the water content became uniform. Solid culture was carried out at 4 ° C. for 4 days. After completion of the culture, wheat bran was washed with 100 g of sterilized water and centrifuged at 16,000 × g for 30 minutes at 4 ° C. to remove insoluble matters. As a result of evaluating the enzyme activity by the maltose decomposition activity described in the above <Analysis method>, 53 U of activity was obtained per 1 g of medium raw material.

<Reference Example 2: Purification of ATCC 10254 strain-derived transferase>
The culture solutions 1.8 L and 9,700 U obtained by the method of Reference Example 1 were concentrated, and the following four stages of chromatographic separation were performed.
(1) Anion exchange chromatography (first time): The culture solution was replaced with 20 mM sodium acetate buffer pH 5.5 by ultrafiltration, and the solution passed through a 0.2 μm filter was used as the enzyme stock solution. The separation resin was TOYOPEARL DEAE-650M (manufactured by Tosoh Corporation), and the amount of resin (hereinafter referred to as CV) was 100 mL.
Using 20 mM sodium acetate buffer pH 5.5 as the first buffer, 0 → 0.4M
Elution was performed with a linear concentration gradient (8 CV) of sodium chloride, and the fraction containing maltose-degrading activity described in <Analysis method> was collected.

  (2) Anion exchange chromatography (second time): The fraction obtained in (1) is collected, replaced by the initial buffer solution having the same composition as (1) by ultrafiltration, and again separated by cation exchange resin. Went. Elution was performed with a linear concentration gradient of CV = 25 mL, 0 → 0.25 M sodium chloride (10 CV), and fractions containing maltose-degrading activity were collected.

  (3) Gel filtration chromatography: The fraction obtained in the step (2) was collected, and then purified by gel filtration chromatography. Separation was performed with a column connecting two HiLoad 16/60 Superdex 200 pg (manufactured by GE Healthcare Japan) and one HiLoad 16/60 Superdex 75 pg (manufactured by GE Healthcare Japan). The sample was replaced with 20 mM acetate buffer pH 5.5 containing 0.2 M sodium chloride by ultrafiltration and simultaneously concentrated to 1 mL. It was applied to a column equilibrated with a buffer having the same composition, and a fraction containing maltose-degrading activity was recovered from the obtained fraction.

(4) Isoelectric focusing (chromatographic focusing): The fraction obtained in the step (3) was collected and chromatofocused. MonoP 5/200 GL (manufactured by GE Healthcare Japan) was used as the column. Chromatography with a gradient of pH 5.5 to 3.5 using 0.025M histidine-hydrochloric acid buffer pH 5.5 as an initial buffer and Polybuffer 74-hydrochloric acid buffer pH 3.5 (manufactured by GE Healthcare Japan) as an elution buffer. The fraction containing 2 U of maltose-degrading activity was collected.
The obtained fraction was evaluated for purity by Native-polyacrylamide gel gel electrophoresis (PhastSystem, manufactured by GE Healthcare Japan, hereinafter referred to as Native-PAGE), and the results are shown in FIG. 1a (left). It was a single band of 97,000 daltons.

<Reference Example 3: Properties of transferase from ATCC 10254 strain>
The result obtained by analyzing the enzyme obtained by the method of Reference Example 2 by SDS-polyacrylamide gel electrophoresis (PhastSystem, manufactured by GE Healthcare Japan, hereinafter referred to as SDS-PAGE) is shown in FIG. 1a (right side). Show. Two molecules having molecular weights of 48,000 and 59,000 were detected. Compared with the results by Native-PAGE, this enzyme was considered to have a heterodimeric structure.
Moreover, it analyzed by isoelectric focusing (PhastSystem, GE Healthcare Japan), and the isoelectric point was about 4.9-5.5 (center value is 5.2). Since proteins are modified with sugar chains and the like, they are considered to show a wide distribution.

<Optimum temperature and pH>
The effects of temperature and pH on the enzyme activity were examined according to the method for measuring maltose decomposition activity described in <Analysis method> above. The evaluation of the influence of pH was carried out using Briton-Robinson buffer instead of acetate buffer.
The results are shown in FIG. 2 (effect of temperature) and FIG. 3 (effect of pH). The optimum temperature of the enzyme was pH 4.0 at a reaction of pH 4.0 for 10 minutes, and the optimum pH was 3.5 at a reaction of 50 ° C. for 10 minutes.

<Temperature stability, pH stability>
The stability of temperature and pH with respect to the enzyme activity was examined according to the method for measuring maltose degrading activity described in <Analysis method> above. The pH stability was evaluated using Briton-Robinson buffer instead of acetate buffer.
For temperature stability, an enzyme solution (20 mM acetate buffer, pH 4.0) was held at each temperature for 30 minutes, cooled with ice water, and then the remaining enzyme activity was evaluated. The pH stability was determined by maintaining the enzyme solution (20 mM Briton-Robinson buffer solution at each pH) at 4 ° C. for 24 hours, adjusting the pH to 4.0, and then evaluating the remaining enzyme activity.
The respective results are shown in FIG. 4 (temperature stability) and FIG. 5 (pH stability). As for the temperature stability of this enzyme, 95% or more of the initial activity remained at 65 ° C, and 90% or more remained at 70 ° C. The pH stability was in the range of 3.0 to 5.0.
<Substrate specificity>
The substrate specificity of this enzyme was evaluated under the conditions of 50 ° C. and pH 4.0. It shows in Table 18 below.

It works well on maltose, disaccharides such as cordobiose, nigerose and sucrose, and malto-oligosaccharides having a polymerization degree of 2 to 7, amylose, soluble starch, etc. to produce glucose. Nigerose, which is a disaccharide having an α1,3 bond, is the best substrate.
With respect to isomaltoligosaccharides having a degree of polymerization of 2 to 4, panose, α-cyclodextrin, amylose and glycogen, the activity is lowered. A slight reaction was observed for paranitrophenyl α-glucoside, methyl α-glucoside, and γ-cyclodextrin. No reaction was observed with trehalose or β-cyclodextrin.

<Influence of metal ions>
The influence of each metal ion of this enzyme was carried out according to the method for measuring maltose degrading activity described in <Analysis method> above. Table 19 shows the results obtained by measuring various ions added to the reaction system.
The activity of this enzyme is inhibited by zinc ions and copper ions. EDTA did not affect the maltose degradation activity of this enzyme.

<Amino acid sequence>
The amino acid sequence contained in the transferase was analyzed by the following method. A protein band was cut out from the SDS-PAGE gel, and the S—S bond in the protein molecule was cleaved by reductive alkylation treatment, followed by peptide fragmentation by trypsin digestion to prepare an analytical sample. LC / MS (high performance liquid chromatograph mass spectrometer, Agilent 1100 series, manufactured by Agilent Technologies) was used. As a result of comparison with the sequence of Aspergillus niger CBS 513.88 strain, it was predicted that the following sequence was contained from the band having a molecular weight of 59,000.
Sequence I: LLVEYQT DERLHVMIYDADEVYQVPESVLPR,
Sequence II: TWLPDDPYVYGLGEHSDPMR,
Sequence III: IPLETMWTDIDYMDKR,
Sequence IV: VFTLDPQR
Moreover, it was estimated that the following sequences were contained from the band of molecular weight 48,000.
Sequence V: WASLGAFYTFYR

The comparison result between the above sequence and the known enzyme amino acid sequence is shown in FIG. 1b. It is predicted that there are multiple types of α-glucosidase produced by Aspergillus niger (Nature Biotechnology, Vol. 25, p. 221, 2007). The amino acid sequence described in FIG. 1b is one of them (Gene agdB, Accession no. An01g10930).
The transferase used in the present invention is the sequence indicated by the underlined portion in FIG. 1b, that is, the sequence of Gene agdB at five positions of amino acid numbers 64-95, 146-165, 295-310, 312-319, 619-630. Since there is a match with (SEQ ID NO: 6), there is a high possibility that it is Gene agdB. Until now, it has not been found that Gene agdB has substrate characteristics, transfer characteristics, heat resistance characteristics, and the like, as in the enzyme used in the present invention.

As a comparative sequence, known α1,4, α1,6-linked oligosaccharide-producing enzyme (α-glucosidase derived from Aspergillus niger, Gene agdA, Accession no .: ang_An04g06920, Agricultural and Biological Chemistry, Vol. 1, Volume 91, p. 3, α1,6-linked oligosaccharide-forming enzyme (Lactobacillus johnsonii-derived α-glucosidase, Gene ljag31, Biochimie 91, 1434, 2009), α1,2, α1,3-linked oligosaccharide-forming enzyme (buckwheat-derived α-glucosidase) Kai 2002-65273, Agricultural and Biological Chemistry Vol. 32, 92 9 page 1968), the amino acid sequence was compared with that of the above thermostable transferase. Moreover, the heat resistance of the enzyme having these comparative sequences is lowered to 40% or less at 65 ° C. for 15 minutes with the enzyme derived from Aspergillus niger. The enzyme derived from Lactobacillus johnsonii decreases to 20% or less at 55 ° C. for 10 minutes. In the buckwheat-derived enzyme, no activity remains at 65 ° C. for 10 minutes.
From the above, heat resistance was inferior.

  α1,3-linked oligosaccharide-producing enzyme: Acrenium sp-derived α-glucosidase (described in JP-A-7-59559 and JP-A-11-9276, strain name: S4G13) has a sequence similar to sequences III, IV, V However, each sequence III, IV, V had two differences from the thermostable transferase. This α-glucosidase produces carbohydrates having α1,3 bonds and α1,4 bonds, but almost no α1,2 bond transition is observed, and the heat resistance is poor as described above (at 55 ° C. for 30 minutes). Activity reduced to 65%).

Other α1,3-linked oligosaccharide-producing enzymes: α-glucosidase derived from Acremonium implicatum (Biochimica et Biophysica Acta Vol. 1700, page 189 2004 and JP 2004-173650 A), similar to sequences III, IV, V However, there were two differences in the sequences III and IV from the transferase of the present invention and one difference in the sequence V. This α-glucosidase produces carbohydrates with α1,3 bonds and α1,4 bonds, but there is almost no α1,2 bond transition, and heat resistance is poor (described as having no activity at 60 ° C for 15 minutes) ).
Therefore, it can be seen that a substance matching any one or more of the above sequences I to V has heat resistance and can produce a novel sugar composition.
The sequence of α1,2 and α1,3-linked oligosaccharide-producing enzyme (α-glucosidase derived from Paecilomyces lilacinus, JP-A No. 2003-169665) was not disclosed.

<Reference Example 4: Transfer of maltose by purified enzyme>
The purified enzyme obtained by the method of Reference Example 2, 0.5 mL (0.7 U / mL) was allowed to act on 1 mL of 45% maltose (final concentration 30%), and the reaction was started at 65 ° C.
After the reaction for a predetermined time, the enzyme was inactivated by heating in a boiling bath for 10 minutes.
The polymerization degree distribution in the reaction solution was analyzed by gel filtration chromatography described in <Analysis method> above. Further, the composition of isomers of disaccharides and trisaccharides was analyzed by the post-column method described in <Analysis Method>, and the composition of each component was calculated from two types of HPLC. Chromatograms of two types of HPLC analysis 48 hours after the start of the reaction are shown in FIGS. 6a and 6b.

The molecular weight distribution 48 hours after the start of the reaction was 25.2% monosaccharide, 29.5% disaccharide, 20.8% trisaccharide, 24.5% over 4 sugars. Analysis of isomers in 2,3 sugars revealed that 15.6% of low molecular weight carbohydrates having α1,2 bonds (hereinafter referred to as α1,2 linked oligosaccharides) and low molecular weight carbohydrates having α1,3 bonds ( Hereinafter, α1,3-linked oligosaccharide) was 14.9%. Among the cord oligosaccharides, 11.2% of cordobiose, which is a disaccharide, and 4.4% of cordobiosyl glucose, which is a trisaccharide, were obtained. Among the nigerooligosaccharides, disaccharide nigerose was 8.5%, trisaccharide nigerotriose was 2.5%, and nigerosyl glucose was 3.9%. The ratio of α1,2 linked oligosaccharide + α1,3 linked oligosaccharide in the disaccharide and trisaccharide components was 61%.
In addition, the dietary fiber content was calculated according to the method described in <Analysis Method> above, and was 31%. In the reaction liquid, the component having 3 or more sugars was 45.3%, and it was considered that 68% of the components having 3 or more sugars contained carbohydrates having α1,2 and α1,3 bonds.

  Changes with time up to 72 hours after the reaction is opened are shown in FIGS. 7a and 7b. In the polymerization degree distribution, the maltose as a substrate decreases and trisaccharide components are produced, and the polymerization degree further increases. Along with the synthesis of trisaccharide and tetrasaccharide or higher components, increases in α1,2 linked oligosaccharides and α1,3 linked oligosaccharides are observed, and the dietary fiber content also increases. Therefore, it can be seen that the amount of carbohydrates having α1,2 bonds and α1,3 bonds accumulates as transfer products with the reaction time.

<Reference Example 5: Transfer product of maltose by crude enzyme>
The crude enzyme 0.5 mL (4.0 U / mL) obtained by the method of Reference Example 1 was allowed to act on 1 mL of 45% maltose (final concentration 30%), and the reaction was started at 65 ° C. 72 hours after the start of the reaction, the enzyme was deactivated by heating in a boiling bath for 10 minutes. The polymerization degree distribution in this reaction solution, disaccharide, and trisaccharide isomer analysis were performed in the same manner as in Reference Example 4. .
The results are shown in Table 20 below. As in Reference Example 4, α1,2-linked oligosaccharides and α1,3-linked oligosaccharides were produced, and the α1,3-linked oligosaccharides and α1,3-linked oligosaccharides in the disaccharide and trisaccharide components of the reaction solution were observed. The total percentage was 52%.
As a result of calculating the dietary fiber content, it was 31%, and it was considered that 61% of the components of the reaction solution having 3 or more sugars contained carbohydrates having α1,2, α1,3 bonds.

<Reference Example 6: Preparation of other Aspergillus-derived transferases>
Aspergillus niger van Tieghem var. niger fsp. Henbergii Blochwitz ex Al-Musallam (NBRC [NITE Biological Resource Center] 4043) and Aspergillus awori (ATCC14331) were used in the same manner as in Reference Example 1 for main culture using a liquid culture medium and a solid culture medium. Culture extracts having 41 U and 35 U activities per gram were obtained.
From each extract, the anion exchange chromatography and the gel filtration chromatography were performed twice according to the method of Reference Example 2 to obtain a partially purified enzyme. The properties of these enzymes were evaluated according to the method described in Reference Example 3. The results are summarized in Table 21 together with the ATCC10254 strain.

  Using these partially purified enzymes, according to Reference Example 5, a transfer reaction was carried out when maltose was used as a substrate, and the composition of the product was analyzed. As a result, α1,2 bond as in the case of the enzyme derived from ATCC10254, It was confirmed that carbohydrates having α1,3 bonds were produced.

<Reference Example 7: Transferred product of maltopentaose by purified enzyme>
0.5 mL (0.7 U / mL) of the purified enzyme obtained by the method of Reference Example 2 was allowed to act on 1 mL of 45% maltopentaose (final concentration 30%), and the reaction was started at 65 ° C. The enzyme was inactivated by heating for 10 minutes in a boiling bath 72 hours after the start of the reaction. The polymerization degree distribution in this reaction solution, isomer analysis of disaccharide and trisaccharide were performed using the same method as in Reference Example 4, and the composition analysis of each component was performed. The analysis results are shown in Table 22.
As in Reference Example 4, α1,2-linked oligosaccharide and α1,3-linked oligosaccharide were produced, and the ratio of α1,2-linked oligosaccharide and α1,3-linked oligosaccharide in the disaccharide and trisaccharide components of the reaction solution The total was 54%. As a result of calculating the dietary fiber content, it was 44%, and it was considered that 90% of the components of the reaction solution having 3 or more sugars contained carbohydrates having α1,2, α1,3 bonds.

<Reference Example 8: Simultaneous reaction of liquefied liquid transfer product, α-amylase, and debranching enzyme with purified enzyme>
The transfer reaction using a polymeric α-glucan as a substrate was carried out as follows. 100 g of a 30 wt% corn starch liquefied solution of DE10 was adjusted to pH 6.0. At a liquid temperature of 65 ° C., the purified enzyme obtained by the method of Reference Example 2 was 0.7 U per gram of solid content, α-amylase (Termamyl 200L, Novozymes) vs. solid content of 0.01% by weight, debranching enzyme ( (Christase PLF, manufactured by Amano Enzyme Co.) was added to the solid content of 0.1% by weight and reacted for 72 hours.
The enzyme was inactivated by heating for 10 minutes in a boiling bath 72 hours after the start of the reaction. The polymerization degree distribution in this reaction solution, isomer analysis of disaccharide and trisaccharide were performed using the same method as in Reference Example 4, and the composition analysis of each component was performed. The analysis results are shown in Table 23.
As in Reference Example 4, α1,2-linked oligosaccharides and α1,3-linked oligosaccharides were produced, and the α1,3-linked oligosaccharides and α1,3-linked oligosaccharides in the disaccharide and trisaccharide components of the reaction solution were observed. The total percentage was 57%.
As a result of calculating the dietary fiber content, it was 55%, and it was considered that 77% of the components of the reaction solution having 3 or more sugars contained carbohydrates having α1,2, α1,3 bonds.

<Reference Example 9: Transfer of liquefied liquid by crude enzyme, simultaneous reaction with α-amylase and debranching enzyme, separation by gel filtration column>
The transfer reaction using a polymeric α-glucan as a substrate was carried out as follows. 100 g of 30% (W / W) corn starch liquefied liquid of DE10 was adjusted to pH 6.0. The purified enzyme obtained by the method of Reference Example 2 at a liquid temperature of 65 ° C. was subjected to 4.0 U per 1 g of solid content, α-amylase (Termamyl 200L, manufactured by Novozymes), 0.01% by weight of solid content, debranching enzyme ( (Christase PLF, manufactured by Amano Enzyme Co., Ltd.) was added in an amount of 0.1% by weight of solid content and reacted for 72 hours.
The enzyme was inactivated by heating for 10 minutes in a boiling bath 72 hours after the start of the reaction. The polymerization degree distribution in this reaction solution, isomer analysis of disaccharide and trisaccharide were performed using the same method as in Reference Example 4, and the composition analysis of each component was performed. The analysis results are shown in Table 24.
As in Reference Example 4, α1,2-linked oligosaccharides and α1,3-linked oligosaccharides were produced, and the α1,3-linked oligosaccharides and α1,3-linked oligosaccharides in the disaccharide and trisaccharide components of the reaction solution were observed. The total percentage was 53%. As a result of calculating the dietary fiber content, it was 55%, and it was considered that 74% of the components of the reaction solution having 3 or more sugars contained carbohydrates having α1,2, α1,3 bonds.

  The sample obtained by measuring the dietary fiber content was purified and concentrated to obtain a processed product having a solid content of 1 g. The sample was added to a gel filtration column (φ6 cm × 100 cm) packed with gel filtration resin Bio-Gel P2 (Bio-Rad), and chromatographic separation was performed to recover a trisaccharide or higher component. Purification, concentration, and lyophilization were performed by a conventional method to obtain 0.2 g of a fraction.

  In order to confirm the binding mode contained in the fractionated sample, the methylation analysis described in <Analysis method> was performed. The results are shown in Table 25 below. It can be seen that 1,3 bonds and 1,3 bonds are contained in the components of three or more sugars calculated as dietary fiber content.

<Reference Example 10: Transfer of liquefied liquid by crude enzyme (derived from ATCC 14331), simultaneous reaction with α-amylase and debranching enzyme, separation by simulated moving bed chromatographic separation device>
Using the crude enzyme derived from Aspergillus awamori (ATCC 14331) obtained by the method of Reference Example 6, a reaction using a polymeric α-glucan as a substrate was performed by the method described in Reference Example 9. The polymerization degree distribution in this reaction solution, isomer analysis of disaccharide and trisaccharide were performed using the same method as in Reference Example 4, and the composition analysis of each component was performed. The analysis results are shown in Table 26.
As in Reference Example 4, α1,2-linked oligosaccharides and α1,3-linked oligosaccharides were formed, and the ratio of α1,2-linked oligosaccharides and α1,3-linked oligosaccharides in the disaccharide and trisaccharide components of the reaction solution The total was 44%. As a result of calculating the dietary fiber content, it was 55%, and it was considered that 65% of the components of the reaction solution having 3 or more sugars contained carbohydrates having α1,2, α1,3 bonds.

30 wt% and 50 kg of the resulting reaction solution were adjusted to pH 5.0, α-amylase (Termamyl 200L, Novozymes) was 3% by weight in solids, and glucoamylase (AMG, Novozymes) was in solids 8 The reaction was carried out at 60 ° C. for 1 hour after addition by weight. The sample after enzymatic degradation was purified and concentrated to obtain a processed product having a solid content of 10 kg.
The enzyme-treated product is separated by a continuous chromatographic separation device (Tresone, manufactured by Organo) packed with a salt-type strongly acidic cation exchange resin (FX1040, manufactured by Organo) to fractionate components of 3 or more sugars. Purification, concentration, and spray drying were performed to obtain a fraction having a solid content of 2 kg. The sugar composition of the fractionated samples was 0.0, 0.0, 27.1, and 72.9% for monosaccharide, disaccharide, trisaccharide, and tetrasaccharide or more. Further, the dietary fiber content was calculated according to the method described in <Analysis method>, and as a result, it was 93%.
In order to confirm the binding mode contained in the obtained sample, methylation analysis was performed in the same manner as in Reference Example 9. The results are shown in Table 27 above. The chromatographic fraction contains 1,2 bonds,
It can be seen that there are 1,3 bonds.

  The sugar composition of the present invention can be used as an additive in a wide range of fields such as pharmaceuticals, cosmetics and livestock feed as well as foods and beverages such as beverages and confectionery. The sugar composition of the present invention is easily dispersible in water, and can be used as a water-soluble dietary fiber that has a high utility value as an additive for beverages and foods.

Claims (8)

A sugar composition containing dietary fiber having glucose as a constituent sugar,
A sugar composition in which the numerical values of x, y, and z in the following formulas are 0.20 ≦ x ≦ 0.75, 0.25 ≦ y ≦ 0.80, and z ≦ 0.30, respectively.
x: Content of dietary fiber with a degree of polymerization of 3 and 4 with glucose as a constituent sugar (g) / Total content of dietary fiber with glucose as a constituent sugar (g)
y: Content of dietary fiber with a degree of polymerization of 5 to 9 using glucose as a constituent sugar (g) / Total content of dietary fiber containing glucose as a constituent sugar (g)
z: Content of dietary fiber with a degree of polymerization of 10 or more with glucose as a constituent sugar (g) / Total content of dietary fiber with glucose as a constituent sugar (g)   2. The sugar composition according to claim 1, wherein the total content of dietary fiber containing glucose as a constituent sugar is 5% by weight or more based on the total weight of the sugar containing glucose as a constituent sugar.   The sugar composition according to claim 1 or 2, wherein the sugar composition is produced by allowing a transferase to act on a sugar raw material.   The transferase acts on any one or more sugar raw materials selected from the group consisting of α-glucan, α-glucooligosaccharide, and glucose, and a carbohydrate having an α1,2 glucoside bond, α1 The saccharide composition according to claim 3, which is a transferase derived from the genus Aspergillus, which produces a saccharide having a glucoside bond and is capable of enzymatic reaction at a temperature of 65 ° C or higher.   The sugar composition according to claim 3 or 4, wherein the sugar raw material is a partially decomposed starch obtained by partially hydrolyzing starch with an enzyme or acid other than the transferase.   The sugar composition according to claim 5, wherein the enzyme is α-amylase.   The food / beverage products containing the sugar composition of any one of the said Claims 1-6.   The said food / beverage products are fermented food / beverage products, The food / beverage products of Claim 7.