KR0131134B1 - Preparation process of isomalto oligo saccharide in hign yield and high purity - Google Patents

Abstract

Isomalto oligosaccharide with high yield rate and high purity is produced by (1) applying sugar-transition enzyme to sugar solution liquidizing and sugaring dextrin and then obtaining isomalto oligosaccharide solution, (2) adding yeast to the above oligosaccharide solution and cultivate the yeast to be fermented about 25-45 deg C for 2-7 days, and eliminating oligosaccharide in the straight chain type such as dextrose included in isomalto oligosaccharide solution, maltose, maltotriose or the like, (3) separating yeast from the above culture fluid.

Description

Process for producing high yield and high purity isomaltooligosaccharide

The present invention relates to a method for producing a high yield, high purity isomalto oligosaccharides, which will be described in more detail. The isomaltooligosaccharide solution obtained by adding a sugar transfer enzyme to the liquefied and saccharified sugar solution is fermented with yeast and contained in isomaltooligosaccharide. The present invention relates to a method for producing high yield and high purity isomalto oligosaccharides by removing straight oligosaccharides such as glucose and maltose.

In general, isomaltooligosaccharide is used as a sweetener for various foods and commercial drinks as a substitute for sugar or fructose. As a method of producing such isomaltooligosaccharide with high purity, a sugar transfer enzyme derived from microorganism is applied to a sugar solution liquefied and saccharified from starch to obtain isomaltooligosaccharide, and a method of increasing the purity using chromatography is used. According to this method, however, maltose and maltotriose, which are composed of α-1.4 bonds, are present in the isomalto oligosaccharide solution. Thus, low molecular weight isomalto oligosaccharides having similar molecular weights, i.e., panos, isomaltose, or isomaltriotriose, etc. It is not easily separated by chromatography, and thus has a disadvantage in that it is difficult to raise the purity to 90% or more.

Korean Patent Application No. 93-17510 filed by Applicant filed Sept. 2, 1993 in order to solve the above disadvantages is a method of increasing the separation efficiency by decomposing the linear oligosaccharides into low molecular weight glucose by adding glucoamylase to the isomalto oligosaccharide solution. Presented. However, although the purity can be increased to 90% or more by the above method, the isomaltooligosaccharide is partially decomposed to low molecular weight by glucoamylase, and the isomaltoligosaccharide is released into the glucose fraction during the chromatography fraction, and the yield is lowered to 70% or less. There was a downside.

Accordingly, an object of the present invention is to provide a method for producing high purity isomalto oligosaccharides in high yield by supplementing the above disadvantages.

The method of the present invention for achieving the above object is a sugar transfer enzyme to the liquefied, saccharified sugar solution from the starch to obtain isomalto oligosaccharide solution, and then add the yeast to the oligosaccharide solution at a temperature suitable for the yeast fermentation It consists of culturing for a period of time to remove glucose contained in the isomaltooligosaccharide solution and linear oligosaccharides such as maltose and maltotriose, and to remove the fermented strain from the culture solution.

The method of the present invention is described in more detail below. According to the present invention, the sugar solution liquefied and saccharified from starch is added to obtain sugar isomalto oligosaccharide solution by adding a sugar transfer enzyme, and then yeast is added and cultured for about 2-7 days at about 25-45 ° C. to glucose, maltose or Linear oligosaccharides such as maltotriose are removed, and then yeast is removed using centrifugation or a filtration agent, followed by treatment with activated carbon treatment and ion exchange resin to prepare purified isomaltooligosaccharide.

The raw sugar solution of the sugar transition reaction used in the present invention is a sugar solution liquefied and saccharified in a conventional manner from starch, and isomalto oligosaccharide solution is added with 0.01-1.0% of a sugar transfer enzyme to the sugar solution of about 20% or more solid concentration. It is obtained by reacting for about 15-48 hours at 45-70 ° C, and contains about 40% or more of non-fermentable sugars. The raw sugar solution may be commercially available general sugars such as malt sugar or mortgage, and isomalto oligosaccharides may be commercially available oligosaccharides.

Yeast may be added directly to the isomalto oligosaccharide solution in order to add yeast to the isomalto oligosaccharide, or to prepare more purified oligosaccharide, the isomalt oligosaccharide is separated by chromatography using an ion exchange resin, and then the yeast is separated. It may also be added. The isomaltooligosaccharide may be treated with alpha glucosidase to decompose maltose into glucose so that the yeast is easy to use, and then yeast may be added. By the added yeast, components such as glucose, maltose and maltodriose remaining in the isomaltooligosaccharide solution are fermented and removed to obtain high-purity isomaltooligosaccharide.

The fermentation concentration of the isomaltooligosaccharide prepared as described above is about 10-40%, which is inferior when the fermentation concentration is lower than about 10% and the volume of the fermentation broth becomes inefficient. This is because the fermentation rate is slowed down.

The yeast may be any of yeast such as baker's yeast and brewer's yeast, and may also use yeast itself or immobilized yeast immobilized on an insoluble carrier. When yeast is simply added, it is preferable to add about 0.5-10.0 g of yeast with respect to 100 g of isomaltooligosaccharide solids, and about 0.5-20.0 ml by total volume in the case of immobilized yeast. This is because when the yeast is added to about 0.5g or less, the incubation time is too long, and when it is added to about 10.0g or more, the efficiency decreases.

When using immobilized yeast, any carrier may be used as long as it is a commercially available product capable of adsorbing yeast. In addition, it is better to use immobilized yeast rather than directly using the yeast for fermentation, because immobilized yeast is reused or immobilized yeast is easily purified.

The sugar solution obtained by the above-described process is composed of about 90% or more of isomaltooligosaccharide and a small amount of straight chain oligosaccharide to obtain high-purity isomaltooligosaccharide.

Hereinafter, the method and effect of the present invention will be described in more detail with reference to Examples and Comparative Examples, but the scope of the present invention is not limited to the following Examples.

Preparation Example 1

5L of a solution (33%) suspended in water for maltose solution was adjusted to pH 5.5, 1.5 ml of glycotransferase (Japan, Amanoshi, Transglucosidase) was added and reacted at 60 ° C. for 30 hours to give isomaltooligosaccharide. After obtaining it was maintained for 10 minutes at 90 ℃ deactivated the enzyme. The sugar solution was analyzed by high pressure liquid chromatography (HPLC), and the sugar composition is shown in Table 1 below.

Example 1

The baker's yeast was immobilized with 2.5% sodium alginate to prepare an immobilized carrier having a diameter of 2-3 mm. 100 ml of the sugar solution of Preparation Example 1 was adjusted to 20% concentration, and 30 ml of immobilized carrier was added to the total volume including the void volume, followed by incubation at a temperature of 27 ° C. for 3 days. After the filtration process to separate the immobilized yeast from the culture solution, the sugar composition separated from the immobilized yeast was analyzed by the high-pressure liquid chromatography and shown in Table 1 below. In addition, the total content of isomaltooligosaccharide after yeast culture was compared with that before yeast culture to obtain a yield, which is shown in Table 1 below. The purity of isomaltooligosaccharide was 92.2% and the yield was 95.1%.

Example 2.

1 L of the sugar solution of Preparation Example 1 was desalted in an ion exchange resin and concentrated under reduced pressure so as to have a solid content of 50%. A column having a diameter of 2 cm and a length of 110 cm was filled with a cation exchange resin (Dow Chemical Co., Ltd., Dowex 99 ') and separated by chromatography to obtain an isomaltoligo oligosaccharide fraction. After adjusting the fraction of isomaltooligosaccharide to 30% concentration, 100 ml of the mixture was added, 10 ml of immobilized yeast was added to the total volume, and the temperature was adjusted to 27 ° C., followed by incubation for 2 days. The obtained sugar solution was filtered and analyzed by high pressure liquid chromatography. As a result, the purity of isomaltooligosaccharide was 97.5%, and the yield was 85.3%.

Example 3.

The sugar solution of Preparation Example 1 was adjusted to 20% concentration, 100 ml was taken, and 5 g of brewer's yeast was added thereto, followed by incubation at 30 ° C. for 2 days. The cells were centrifuged and filtered to remove the cells. The resulting sugar solution was decolorized by activated carbon treatment, desalted with an ion exchange resin, and concentrated to give 10 g of a high purity oligosaccharide having a concentration of 75%. The sugar solution thus obtained was filtered and analyzed by high pressure liquid chromatography, and the sugar composition and yield are shown in Table 1 below, and the results of 92.6% purity and 93.0% yield of isomaltooligosaccharide were obtained.

Example 4.

The yeast fixation column was prepared by filling immobilized yeast with 20 ml in a circular column with a water jacket. The temperature of the immobilization column was adjusted to 30 ° C. and the isomalto oligosaccharide solution adjusted to 20% concentration was continuously flowed at a rate of 3 ml per hour. The sugar solution obtained here was analyzed by high pressure liquid chromatography, and the sugar composition and yield are shown in Table 1 below. In this example, the purity of isomaltoligoda was 91.1% and the yield was 95.5%.

Example 5

0.1% of alphaglucosidase was added to 1 L of the sugar solution of Preparation Example 1, reacted at 50 ° C for 48 hours, adjusted to 20% concentration, and 100 ml of this solution was added to 30 ml of immobilized yeast in a total volume and 3 days at a temperature of 27 ° C. Incubated. The culture solution was filtered to obtain a sugar solution from which immobilized yeast was obtained, which was analyzed by high pressure liquid chromatography, and the sugar composition and yield are shown in Table 1 below. As a result, the isomaltooligosaccharide showed a purity of 93.5% and a yield of 94.5%.

Comparative Example 1.

The method was compared with Korean Patent Application No. 93-17510. The method was prepared by adjusting glucoamylase (US, Solvay, Dizyme) to 0.001% of sugar solution 1 in Preparation Example 1 and adjusting the pH to 5.0. After reacting at 60 ° C. for 48 hours, the resultant was desalted in an ion exchange resin and concentrated under reduced pressure to 60%. A column having an inner diameter of 2 cm and a length of 110 cm was filled with a cation exchange resin (Dowex 99, Japan, Dow Chemical Co., Ltd.) and chromatographed to obtain an isomaltole oligosaccharide fraction. The sugar solution obtained at this time was analyzed by high pressure liquid chromatography, and the sugar composition and yield are shown in Table 1 below. The purity of isomaltooligosaccharide was 95.3%, but the yield was 65.0%.

As can be seen from the above examples, by removing the straight chain oligosaccharides such as glucose and maltose by the method of the present invention using the yeast action, isomaltole oligosaccharides can be produced in high purity, and the yield of isomaltooligosaccharide is at least 85. A high effect was obtained above%. In addition, there is an advantage that the immobilized yeast can be reused by immobilizing the yeast cells on the carrier. The high-purity isomaltooligosaccharide prepared in this way has excellent bifidus growth and caries suppression effect, so that it can be used not only for general foods but also for diet foods, beverages, health functional beverages and medicines, and for the purpose of reinforcing functionalities. Can be used.

Claims (9)

Isomalto oligosaccharide solution was obtained by acting a sugar transfer enzyme on the liquefied and saccharified sugar solution from starch, and then yeast was added to the oligosaccharide solution, and incubated at about 25-45 ° C. for about 2-7 days for the yeast to ferment. Method for producing a high-yield, high-purity isomaltooligosaccharide characterized in that the glucose and maltose, maltotriose, and straight chain oligosaccharides contained in the maltooligosaccharide solution are removed, and the yeast is separated from the culture solution. The method of claim 1, wherein the solid content of the sugar solution is about 20% or more. The method according to claim 1, wherein the sugar solution is added to the sugar transfer enzyme 0.01-1.0% and reacted for about 15-48 hours at about 45-70 ℃ to obtain an isomalt oligosaccharide containing at least about 40% non-fermentable sugar A method for producing a high yield, high purity isomaltooligosaccharide characterized by the above-mentioned. The method for producing a high yield, high purity isomaltooligosaccharide according to claim 1 or 2, wherein the sugar solution is maltose or mortgage. The method of claim 1, wherein the yeast is added after adjusting the fermentation concentration of the isomaltooligosaccharide to about 10-40%. 6. The method of claim 5, wherein the yeast is added to the yeast itself or an immobilized yeast immobilized on an insoluble carrier. 2. The high yield and high purity according to claim 1, wherein the isomalto oligosaccharide solution to which the yeast is added is isomaltole oligosaccharide itself or isomaltooligosaccharide high content liquid obtained by chromatography of isomaltole oligosaccharide by chromatography using an ion exchange resin. Method for producing isomaltooligosaccharide The method of claim 1, wherein the yeast isomalto oligosaccharide solution to which the yeast is added by treating with alpha glucosidase so that yeast is easy to use, after decomposing maltose and maltotriose to glucose, characterized in that the addition of yeast Yield, high purity isomaltooligosaccharide manufacturing method. The method according to claim 6, wherein when the yeast is added directly, about 0.5-10.0 g is added to 100 g of isomaltooligosaccharide solution, and when the immobilized yeast is added, about 0.5-20.0 ml is added as a total volume. Method for producing isomalto oligosaccharides.