JPS6030695A - Production of nonfermentable sugar containing highly hygroscopic isomaltose as main component - Google Patents

Abstract

PURPOSE:To produce a nonfermentable sugar containing highly hygroscopic isomaltose as a main component, by treating a maltose-containing liquid with transglucosidase, thereby removing the existing glucose, etc. having poor hygroscopicity. CONSTITUTION:A liquid containing nonfermentable sugar is prepared by treating a maltose-containing liquid with transglucosidase, and the obtained sugar liquid is added with glucoamylase to decompose the existing maltose and maltooligosaccharide to glucose. The resultant mixture composed of nonfermentable sugar and glucose is passed through an alkali metal-type strongly acidic cation exchange resin or an alkaline earth metal-type strongly acidic cation exchange resin to separate the nonfermentable sugar fraction from the glucose fraction. A nonfermentable sugar composition containing >=70% nonfermentable sugar can be produced by this process.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing non-fermentable sugar containing isomaltose, which has good hygroscopic properties, as a main component. Further, L< is to remove glucose, maltose, and maltooligosaccharides that have poor hygroscopic properties from a non-fermentable sugar-containing liquid obtained by allowing transglucosidase to act on a maltose-containing liquid,
The present invention relates to a method for producing non-fermentable sugar containing isomaltose as a main component, which has good hygroscopicity and is characterized by obtaining non-fermentable sugar.

In the present invention, non-fermentable sugars refer to trisaccharides, trisaccharides, tetrasaccharides, etc. having α-1,6 bonds, and specific examples include isomaltose, isomaltotriose, panose, nigerose, isomaltotetraose, Examples include cordibiose.

Until now, sugar has been widely used as a sweetener in foods, but in recent years there has been a tendency to refrain from using sugar as a sweetener that tends to cause tooth decay.

For this reason, starch syrup containing fructose, glucose, or maltose as the main ingredients has gradually come to be used as a sweetener instead of sugar, but fructose and glucose tend to discolor when heated and may not be used in some foods. Although maltose poses no problem in terms of coloring, it has poor hygroscopicity, so foods that use maltose syrup as a sweetener tend to dry out easily and become dull.
Therefore, other sweeteners with good coloring and hygroscopicity have been sought to replace sugar. .

Therefore, the present inventors focused on non-fermentable sugars, which have not been used in the past, and considered applying them to sweeteners. Until now, there have been reports on the production of non-fermentable sugars containing isomaltose as a main component. That is, this is a method for obtaining non-fermentable sugars by allowing transglucosidase produced by the filamentous fungus Aspergillus to act on a maltose-containing solution (Japanese Patent Publication No. 40-27319 and Japanese Patent Application Laid-open No. 56-51.982). However, in these methods, the non-fermentable sugar content (ratio of non-fermentable sugars in total sugars) is only about 50%, and the proportion of glucose, maltose, and maltooligosaccharides in addition to non-fermentable sugars is only about 50%. of about 50%, therefore, it could not be used as a sweetener with good hygroscopicity.

On the other hand, attempts have also been made to separate specific components such as isomaltose from mixtures of non-fermentable sugars.

Separation methods such as paper chromatography or activated carbon column chromatography have been used for a long time. However, these separation means are complicated to use and difficult to scale, and cannot be said to be a method that can be used industrially.

Recently, after allowing transglucosidase to act on a maltose-containing solution, glucoamylase is added to hydrolyze the coexisting oligosaccharides into glucose, and yeast is further added.
Subsequently, a method for obtaining isomaltose by fractionating it using Toyo Pearl HW-40 (trade name, manufactured by Toyo Tsudani Gyo Co., Ltd.) column chromatography has also been reported (Page 216 of the 1981 Japan Agricultural Chemistry Conference Abstracts) . However, this method uses the principle of molecular sieves, which has problems with scale-up, and it is a method that isolates only isomaltose, and the isomaltose obtained by this method has a low recovery rate from maltose, which is the raw material. 25%
The sweetener is weak and expensive, and cannot be used as it is as the sweetener of the present invention.

The present inventors conducted studies to improve the shortcomings of these conventional methods. That is, transglucosidase is applied to a maltose-containing solution to obtain a sugar solution with a non-fermentable sugar content of about 50%, and then the glucose, maltose, and
An attempt was made to separate sugars with poor hygroscopic properties such as malto-oligosaccharides from non-fermentable sugars with good hygroscopic properties. Among the mixed glucose, maltose, and maltooligosaccharides, maltose and maltooligosaccharides are decomposed into glucose by glucoamylase to form a mixture of non-fermentable sugar and glucose, and then the mixture is treated with strong acidic cations. By passing the liquid through an exchange resin, they succeeded in separating the non-fermentable sugar fraction and the glucose fraction. The non-fermentable sugar that is mainly composed of isomaltose obtained in this way is obtained by allowing transglucosidase to act on a maltose-containing liquid and is recovered almost as is, and the resin used can be easily scaled up. Therefore, it is an industrial production method, and the non-fermentable sugar whose main component is isomaltose obtained in this way has a moderate sweetness and is not colored.
The present invention was completed based on the knowledge that it has good hygroscopicity.

The starting material maltose-containing liquid used in the present invention is an aqueous solution of commercially available maltose, or corn starch, potato starch, sweet potato starch, soluble starch, etc., mixed with acid or α-
Either β-amylase alone or a maltose-containing liquid obtained by simultaneously acting β-amylase, pullulanase, and/or isoamylase on the liquefied starch obtained by liquefying it with amylase can be used.

Further, in the method of the present invention, the action conditions for producing non-fermentable sugars by acting transglucosidase on maltose-containing liquid are preferably 40 to 60°C for 15 to 48 hours and pH 4.0 to 6.0. In addition, when transglucosidase is applied to produce non-fermentable sugars, in addition to the above methods, simultaneous addition of β-amylase and transglucosidase, or simultaneous addition of β-amylase, pullulanase and glucosidase to the liquefied starch solution may be used. good.

The strongly acidic cation exchange resin used in the present invention is a sulfonic acid type, for example, Amberlite I R
-116, Amberlight IR41B, Amberlight IR-120B,, XT 1022E, XT
471F (all product names, manufactured by Organo); Diaion 5KIB, Diaion 5K102, Diaion 5K104, Diaion 5KLOG, Diaion 5
KIIO, Diaion 5K112, Diaion 5K
116, Diaion FROI (all trade names, manufactured by Mitsubishi Kasei Corporation), etc. are preferable. And these resins are 6ε before use.
Used as alkali metal type or alkaline earth metal type. In addition, in order to improve the separation of non-fermentable sugars and glucose, the flow rate when passing through the column was set at 5V = 0.1 to 0.5.
A range of is preferred. If it is slower than this, the workability will be poor,
This is because if it is too early, the separation of non-fermentable sugars and glucose will deteriorate.

Furthermore, if the temperature at which the liquid is passed is too low, viscosity will occur and the flow rate will become too slow, so it is preferable to carry out the process at room temperature to 70°C.

The strongly acidic cation exchange resin used in the present invention can efficiently separate non-fermentable sugars and glucose, so that the desired isomaltose with a high non-fermentable sugar content can be obtained just by passing the mixture through it once. Although both main components can be obtained, re-chromatography is performed to further reduce the amounts of glucose, maltose and full-l-oligosaccharides, and increase the non-fermentable sugar content and isomaltose. By re-chromatography, it was possible to separate and obtain non-fermentable sugars with a non-fermentable sugar content of 95% or more and a ratio of isomaltose to non-fermentable sugars of 70% or more.

Furthermore, the present invention uses non-fermentable sugar containing isomaltose as a main component as a sweetener, and for this purpose it is necessary to obtain the non-fermentable sugar fraction as efficiently as possible. That is, it is also possible to pass a mixed solution of non-fermentable sugar and glucose through a strongly acidic cation exchange resin to obtain a non-fermentable sugar fraction using a continuous recycling method. In this method, a mixture of 0.1 to 0.5 volumes of the column volume is passed through the column, recycled, and then arranged in the order of elution: fractions with non-fermentable oligos, 2) isomaltose fraction, (3) isomaltose-rich fraction, ( 4) Glucose lynch fraction, (5
) of the glucose fractions, first obtain both fractions (2), take out the fractions (1) and (5), and send the fractions (3) and (4) to the column again. flow to. At that time, the same amount of sugar solution and water as the volume taken out of the system is added into the system. The order of addition is (3), stock solution, and (4), water. By repeating this process, a sugar solution fraction containing isomaltose as a main component with a non-fermentable sugar content of 70% or more is efficiently obtained.

By appropriately concentrating the non-fermentable sugar solution obtained in this way, a non-fermentable sugar containing isomaltose with good hygroscopicity as a main component can be obtained.

The non-fermentable sugar containing isomaltose as a main component of the present invention has a much weaker Maillard reaction than fructose and glucose, and is not inferior to starch syrup containing maltose as a main component.

Furthermore, in terms of hygroscopicity, it is superior to both maltose and glucose, and in terms of sweetness, it has almost the same sweetness as conventional starch syrup whose main ingredient is maltose. It has been found that the non-fermentable sugar contained as a main component can be satisfactorily used as a sweetener.

The present invention will be specifically explained below with reference to Examples.

In addition, the α-amylase activity used in the present invention, β-
Each method for measuring amylase activity, transglucosidase activity, and glucoamylase activity will be explained below.

α-Amylase activity: 2% barley! ! Add enzyme solution 1- to starch (pH 6,0) 10-, react for 10 minutes at 40°C, and then add 1-1 to 10+
0 of f. Add IN hydrochloric acid to stop the reaction. This liquid 1
- is added to 0.005% iodine solution of 10- and exposed to 660 nm.
Measure the absorbance. The time when the iodine coloring decreases by 1% in 1 minute under these conditions is defined as 1 unit.

β-amylase activity: Add enzyme solution 1- to 9 ml of 0.56% soluble starch solution (pH 5,0) and react at 40°C for 30 minutes. The sugar produced under these conditions was measured using the Fehring-Lehmann-Schul method.
When producing reducing sugar equivalent to 10■ of glucose, 1
Unit.

Transglucosidase activity: 4% alpha methyl D glucoside solution 0.5 mF and 0
．． Add enzyme solution 0.57 to a mixture of 0.02M acetate buffer (pH 5,0) and 0.5-0, and perform an enzyme reaction at 40°C for 60 minutes. The reaction is stopped by adding 0.5-O,IN sodium hydroxide solution. After about 2 hours, 0. Add 0.511 Il of IN salt #&solution to neutralize. Glucose produced by the enzymatic reaction was quantified by the glucose oxidase method, and the activity of producing 1 pg of glucose in 60 minutes at 40° C. was defined as 1 unit.

Glucoamylase activity: The pH of the substrate solution was set to 4.5, and the measurement was performed in exactly the same manner as the β-amylase activity.

Example 1 1000n+n of 25% maltose aqueous solution at pH 5.5
and transglucosidase (manufactured by Ohno Pharmaceutical Co., Ltd., 15
00,000 units/g) by adding 75,000 units to 53
The transfer reaction was carried out at 18°C for 18 hours to produce non-fermentable sugars.

Thereafter, the enzyme activity was stopped by heating at 100°C for 10 minutes, and 500 units of glucoamylase (manufactured by Ohno Pharmaceutical Co., Ltd., 6,000 units/g) was added and reacted at 55°C for 24 hours to coexist. Decomposed maltose and maltooligosaccharides into glucose. The thus obtained mixed liquid consisting of non-fermentable sugar and glucose is concentrated under reduced pressure to a solid content of 50%, and this concentrated liquid is then used as a product of Diaion FROINa type (product name), which is an alkali metal strongly acidic cation exchange resin. name, Mitsubishi Kasei product) Length 200 filled with 21
A non-fermentable sugar fraction was collected by passing the solution through a cm0 column (diameter 4.0Ω) and then passing 4p of water through it.

Thereafter, the non-fermentable sugar fraction was concentrated, and the non-fermentable sugar content was 7.
150 g (solid content 75%) of non-fermentable sugar containing isomaltose as a main component and containing 5% or more was obtained. When the sugar composition of this product was analyzed by paper chromatography, it was found to be 46.8% isomaltose, 14.6% isomaltotriose, 20.6% panose, and 2.4% maltose.
, glucose 8.9%, and other 6.7%.

Example 2 1.2 g of amylase Amano AD-1 (manufactured by Ohno Pharmaceutical Co., Ltd., 10,000 units/g), which is a liquefied amylase, was added to a 30% concentration cornstarch suspension FOA, and DE
After cooling to 55°C, 4 g of Biozyme M (manufactured by Ohno Pharmaceutical Co., Ltd., 4000 units/g) having β-amylase activity was added to the starch liquefied liquid No. 12, and at the same time transglucosidase (manufactured by Ohno Pharmaceutical Co., Ltd.) was added. , 1.5 million units/g
) was added, a transfer reaction was carried out for 18 hours to produce non-fermentable sugars, and then heated at 100°C for 10 minutes to stop the enzymatic action, and glucoamylase (manufactured by Ohno Pharmaceutical Co., Ltd., 6000 units /
g) 2g was added and reacted at 55 degrees for 24 hours to decompose coexisting maltose and maltooligosaccharides into glucose.

The thus obtained mixed liquid 20β consisting of non-fermentable sugar and glucose was concentrated under reduced pressure (solid content 50%), and the concentrated liquid 41
was passed through a strongly acidic cation exchange resin, XT471F-Na type (organo product) (column length: 200cm).
nl, diameter 10c111. resin amount 151), then water io
A is passed through water and fractionated into a non-fermentable oligosaccharide fraction, an isomaltose fraction, an isomaltose-rich fraction, a glucose-rich fraction, and a glucose fraction to obtain an isomaltose fraction and a non-fermentable oligosaccharide fraction. The oligosaccharide fraction and the glucose fraction were discharged from the system, and the isomaltose-rich fraction and the glucose-rich fraction were again supplied to the column. At that time, the same amount of sugar solution and water as the volume taken out of the system was added to the system.

The order of addition was the isomaltose-rich fraction, the stock solution, the glucose-rich fraction, and water. By repeating this for 4 cycles, 6 kg of non-fermentable sugar (solid content 50%) containing isomaltose as a main component was obtained.

The sugar composition of this product was investigated by paper chromatography: 34.9% isomaltose, 11.1% isomaltotriose, 26.1% panose, 3% nigerose.
．． 9%, isomaltotetraose 1.9%, maltose 3.9%, glucose 6.6%, and other 5.9%. This product was further chromatographed using the same column, and the same procedure was performed to separate both non-fermentable sugar fractions. The mixture was then concentrated under reduced pressure to obtain 2.7 kg of non-fermentable sugar containing isomaltose as a main component with a solid content of 75%. The sugar composition of this product was investigated by paper chromatography and found to be 75.8% isomaltose and 5% isomaltotriose.
．． 6%, panose 2.3%, nigerose 3.7%, cordibiose 8.3%, glucose 2.3%, other 2.
It was 0%.

Example 3 Non-fermentable sugar (solid content 75%) containing isomaltose as a main component obtained according to Example 1 was heated at 80°C.
The mixture was heated to dryness under reduced pressure to obtain a dry product.

The thus obtained non-fermentable sugar containing isomaltose as a main component of the present invention, maltose powder, and glucose powder were stored at 20°C for 15 hours at a relative humidity of 79%,
The water absorption rates of each were compared. The results are shown in Figure 1. That is, after 15 days, the water absorption rates of the non-fermentable sugar products containing isomaltose as a main component, maltose powder, and glucose powder of the present invention were 18.0% and 9.9%, respectively.
and 6.8%. As is clear from these results, the non-fermentable sugar containing isomaltose as a main component obtained by the present invention is significantly superior in terms of hygroscopicity compared to flu-1-ose and glucose. .

Example 4 The degree of coloring due to heating was compared between the non-fermentable sugar containing isomaltose as a main component of the present invention obtained according to Example 1, and maltose and glucose. i.e. 30g each
0.2% sodium glutamate was added to each aqueous solution obtained by dissolving 100% of water, and the degree of coloring was measured at a wavelength of 420 nm after heating at 85°C for 150 minutes.

When the absorbance was measured using a cell with a layer length of 100 cranes, it was 0.083 and 0.089, respectively, and the non-fermentable sugar containing isomaltose as a main component of the present invention and maltose showed almost the same degree of coloration.

Example 5 Soy sauce 51,
2 kg of non-fermentable sugar (solid content 75%) containing isomaltose as a main component obtained in Example 1 and 2.5 kg of sugar
kg was added and simmered to obtain clam tsukudani.

This tsukudani was appetizing not only in taste and aroma, but also in color and luster.

Example 6 60 kg of bleached bean paste, 70 kg of sugar, 40 kg of non-fermentable sugar (solid content 75%) containing isomaltose as a main component obtained in Example i, and 150 g of agar were added, and this was placed in an airtight pot. Reduce the vapor pressure to 0 while stirring gently.
．． The pressure was increased to 5 kg/cJ, then the vapor pressure was gradually lowered while stirring, and the mixture was cooled to produce Ogura bean paste.

This product had good flavor and color, and the bean paste bread produced by adding this product did not become loose.

[Brief explanation of drawings]

FIG. 1 is a diagram comparing the hygroscopic properties of powders of non-fermentable sugar, glucose and maltose containing isomaltose as a main component of the present invention at RH79 and 20°C. Patent applicant Amano Pharmaceutical Co., Ltd.

Claims (1)

[Scope of Claims] 1. A non-fermentable sugar-containing liquid is obtained by allowing transglucosidase to act on a maltose-containing liquid, and then glucoamylase is added to decompose maltose and maltooligosaccharides mixed in the sugar liquid into glucose, After forming a liquid mixture consisting of non-fermentable sugar and glucose, the liquid mixture is passed through a strongly acidic cation exchange resin to separate the non-fermentable sugar fraction and the glucose fraction, and then the non-fermentable sugar fraction is separated from the glucose fraction. A method for producing a non-fermentable sugar containing isomaltose as a main component with good hygroscopicity, characterized by obtaining the following: 2. Isomaltose with good hygroscopicity according to claim 1, wherein the strongly acidic cation exchange resin is either an alkali metal type strongly acidic cation exchange resin or an alkaline earth metal type strongly acidic cation exchange resin. A method for producing non-fermentable sugar contained as a main component. 3. Hygroscopicity according to claim 1, wherein the content of non-fermentable sugars separated and obtained by passing a mixture of non-fermentable sugars and glucose through a strongly acidic cation exchange resin is 70% or more. A method for producing non-fermentable sugar containing good isomaltose as a main component.