JPH0579315B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Field of Application] The present invention has a low sweetness and has an effect of preventing aging and improving the shelf life of foods and drinks such as confectionery, and is particularly suitable for use as a sweetener with an anti-caries effect, such as in foods and drinks or medicines. The present invention relates to a method for producing a branched oligosaccharide syrup that can be used. [Conventional technology] In recent years, as people's tastes have diversified, a so-called sweet taste phenomenon has occurred.At the same time, as interest in health has increased, sugar has been avoided and other low-calorie sweeteners have become a hot topic. It started to look like it was climbing. There are a variety of low-calorie sweeteners available, both natural and synthetic, but branched oligosaccharides are indigestible, have low cariogenicity, and have other physiological properties, and are used in foods, drinks, pharmaceuticals, etc. , which can be used in a wide range of applications. Until now, such saccharides have not been industrially produced, and isomaltose and the like have only been supplied as expensive reagents. The present inventors focused on so-called branched oligosaccharides containing α-1,6 bonds of glucose molecules, such as isomaltose, and first explained and applied for an industrial and economical manufacturing method (Patent Application No. −245470). According to the above-mentioned industrial production method invented by the present inventors, a product containing about 40% (solid content) of branched oligosaccharides can be obtained in the form of syrup by utilizing the condensation reaction of saccharides. By further purifying this syrup, the concentration of branched oligosaccharides can be reduced to approximately
It is now possible to obtain highly concentrated syrup with a concentration of up to 90% (based on solids). The branched oligosaccharides contained therein mainly include isomaltose, followed by panose, isomaltotriose, and the like. Such sugar syrup products have hardly ever been available on the market, and it is expected that their uses and demand will continue to expand due to the properties of branched oligosaccharides mentioned above. [Problems to be Solved by the Invention] The present invention aims to more effectively advance the condensation reaction of saccharides in the method for producing branched oligosaccharide syrup invented by the present inventors, and to produce branched oligosaccharide syrup more economically. It's something you get. [Means for Solving the Problems] The present invention uses a glucose aqueous solution having a solid content concentration of 60% or more and a glucose content of 70% or more of the solid content as a substrate,
An enzyme capable of condensing saccharides is applied to this at 55°C or higher to produce branched oligosaccharides in the aqueous solution, and then, if necessary, saccharides other than branched oligosaccharides are further separated and removed to form a branched oligosaccharide syrup. This is the manufacturing method. What is the branched oligo here?
It is a general term for oligosaccharides having α-1,6 bonds of glucose molecules, such as isomaltose, panose, and isomaltotriose. As the enzyme having the action of condensing sugars used in this method, glucoamylase is usually selected from the viewpoint of easy availability and ease of use. Examples of glucoamylase include those originating from the genus Aspergillus, such as Aspergillus niger (for example, AMG 300L manufactured by Novo), and those originating from microorganisms of the genus Rhizopus (for example, Sumitym, manufactured by Shin Nihon Kagaku Co., Ltd.). When these enzymes are used in their most common use, the starch hydrolysis reaction for glucose production, the enzymes are activated at temperatures exceeding 60°C for Aspergillus niger glucoamylase and 55°C for Rhizopus glucoamylase. Deactivation occurred, making it practically unusable at higher temperatures. However, the present inventors surprisingly discovered that when these enzymes are made to act on a highly concentrated sugar solution, the enzymes do not deactivate even if the temperature exceeds the above range, but rather the reaction is accelerated. In other words, as shown in Fig. 1, for example, Aspergillus
Glucoamylase of niger origin (AMG manufactured by Novo)
300L), the enzyme is not inactivated even at high temperatures of 60°C or higher, and as the temperature is further increased, the production rate of branched oligosaccharides actually accelerates. Enzyme deactivation occurs at temperatures above 80°C and with reaction time. The same is true for glucoamylase originating from microorganisms of the genus Rhizopus.
At temperatures above the rate of branched oligosaccharide production increases,
It finally begins to lose its activity at temperatures above 80°C. Such behavior of the enzyme in the present invention has not been predicted in the past, and the industrial benefits of utilizing this behavior are large. In other words, it brings about many effects such as increasing the reaction rate by raising the reaction temperature, improving workability by reducing the viscosity of the liquid, and preventing undesirable side reactions by suppressing the actions of other enzymes. It is. The amount of enzyme added is approximately 0.05 to 3.0% (based on solid content).
The reaction PH is between 4.0 and 6.0, and these conditions may depend on the usual usage of these enzymes. The glucose aqueous solution used for substrate loss has a solid content concentration.
60% or more is essential, and as the solid content concentration increases, the amount of branched oligosaccharides produced increases. When the solid content concentration is 85%, the amount of branched oligos produced reaches 60% or more (relative to solid content). Furthermore, as the solid content concentration increases, the heat resistance of the enzyme improves. As mentioned above, it begins to lose its activity when the temperature exceeds 80°C. The solid content of the substrate preferably contains 70% or more of glucose, and may also contain other disaccharides, trisaccharides, etc. Examples of such substrates that can be used practically include glucose crystal dissolution solution, purified glucose solution, etc., as well as nectar after collecting glucose crystals, fructose from isomerized sugar solution, etc., which are conventionally considered by-products in glucose production. Examples include roughinate after separating. These can be used as raw materials for the present invention as they are, or after being adjusted to contain predetermined solid content and glucose content. The sugar solution thus obtained is a mixed sugar solution containing branched oligosaccharides such as isomaltose, panose, and isomaltotriose, as well as glucose, maltose, and the like. This can be used as it is as a branched oligosaccharide syrup, or, if necessary, can be further processed by the following method to obtain a branched oligosaccharide syrup with a higher concentration. (1) A method of concentrating and slowly cooling a sugar solution to crystallize and remove glucose, etc. (2) A method of adding salt to a sugar solution to crystallize and remove glucose-salt double salt crystals. (3) A method in which an organic solvent such as alcohol or acetone is added to a sugar solution to precipitate and remove sugars other than branched oligosaccharides. (4) A method of removing sugars other than branched oligosaccharides using molecular fractionation chromatography such as an activated carbon column or gel filtration, or ion exchange chromatography using an ion exchanger. (5) A method of assimilating sugars other than branched oligosaccharides by allowing yeast to act on a sugar solution. [Example] Example 1 Commercially available glucoamylase originating from Aspergillus niger (manufactured by Novo, AMG 300L) was added to a high-purity glucose solution (DE97.5, manufactured by Showa Sangyo Co., Ltd.) with a concentration of 75%.
was added at a solid content of 0.6% (v/w) and reacted for 72 hours at a pH of 4.5 and a reaction temperature of 60, 65, 70, 75, and 80°C to obtain the reaction products shown in the table below. Ta.

[Table] Not analyzed.
Example 2 A commercially available glucoamylase originating from the genus Rhizopus (Sumizyme, manufactured by Shin Nihon Kagaku Co., Ltd.) was added to a high-purity glucose solution (DE97.6, manufactured by Showa Sangyo Co., Ltd.) at a concentration of 75% with a solid content of 0.9% (solid content: 0.9%). w/w), the pH was set to 4.8, and the reaction temperature was
The reaction was carried out at temperatures of 55, 60, 65, 70, and 75°C for 120 hours to obtain the reaction products shown in the table below.

【table】

[Table] Not analyzed. Analysis was performed using high performance liquid chromatography.
Example 3 The following four types of substrates were prepared at a concentration of 75%, and commercially available glucoamylase (manufactured by Novo, AMG 300L) was used.
0.6% (v/w) was added and reacted at 70°C and PH4.5 for 120 hours to obtain a branched oligosaccharide syrup as shown in the table below. (1) Crystalline glucose solution (glucose purity 99.8%) (2) Purification Produced glucose solution (glucose purity 95.0%) (3) Honey obtained by separating the crystals (Glucose purity 92.1%) (4) Roughinate from chromatographic separation of isomerized sugar syrup (glucose purity 83.6%)

[Table] None.
Example 4 Remaining honey after removing glucose crystals (Showa Sangyo Co., Ltd.)
commercially available glucoamylase (manufactured by Novo, AMG) originating from Aspergillus niger.
300L) was added at a solid content of 0.5% and reacted at pH 4.5 and 70°C for 96 hours to obtain the products shown in the table below.

[Table] After this reaction product was purified using activated carbon and ion exchange resin and concentrated to a concentration of 60%, it was purified using a styrene-based strongly acidic Na-type cation exchange resin (Amberlite).
Monosaccharides were mainly removed by partition chromatography using a column packed with CG-20) to obtain the following highly pure branched oligosaccharide syrup.

[Table] Example 5 Hydrous crystalline glucose (manufactured by Showa Sangyo Co., Ltd.) was dissolved to adjust the concentration to 60, 70, 75, 80, and 85%, and commercially available glucoamylase originating from Aspergillus niger (manufactured by Novo) was dissolved. , AMG 300L) with a solid content of 1.0%
(v/w) and reacted at PH4.5 and 75°C for 120 hours.
The reaction products shown in the table below were obtained.

【table】

[Table] represents the content of.
[Effects of the Invention] The present invention provides an industrial method for efficiently producing branched oligosaccharide syrup. The branched oligosaccharide syrup obtained in this way has a refreshing sweetness that is lower than sugar, has anti-aging effects, and improves shelf life, so it has a wide range of uses as a sweetener or flavor improver for various foods. be. Furthermore, since it has an anti-caries effect, it can be widely used as a sweetener for the purpose of preventing dental caries. In addition, since branched oligosaccharides have the effect of inhibiting the crystallization of sugars, they can also be used as a crystallization inhibitor by adding a small amount to a sugar solution such as sugar, glucose, high fructose sugar, or maltose.

[Brief explanation of the drawing]

Figure 1 shows the production of branched oligosaccharides versus reaction time at each reaction temperature when 0.6% glucoamylase (based on solid content) originating from Aspergillus niger is applied to a substrate with a solid content concentration of 75% (glucose 95%). It is a graph showing the amount.

Claims (1)

[Scope of Claims] 1. Glucose in solid content with a solid content concentration of 60% or more
A method for producing a branched oligosaccharide syrup by a high-temperature condensation reaction, which comprises causing an enzyme having a saccharide condensation action to act on a 70% or more glucose aqueous solution at 55°C or higher to produce a branched oligosaccharide in the aqueous solution. . 2. The method for producing a branched oligosaccharide syrup according to claim 1, wherein the enzyme having the action of condensing saccharides is glucoamylase derived from a microorganism of the genus Aspergillus. 3. The method for producing a branched oligosaccharide syrup according to claim 1 or 2, wherein the enzyme having the action of condensing sugars is glucoamylase derived from a microorganism of the genus Rhizopus.