JPS619266A - Preparation of sweetener to promote multiplication of lactobacillus bifidus from molasses of sugar beet - Google Patents

Abstract

PURPOSE:The titled sweetener comprising raffinose and an oligosaccharide mainly, capable of multiplicating useful bacteria in the intestines even if human takes it, by converting sucrose in molasses of sugar beet with a specific enzyme into a fructooligosaccharide, treating it with an ion exchange resin. CONSTITUTION:Molasses of sugar beet containing raffinose is treated with fructosyltransferase contained in immobilized mold of Aureobasidum pullulans AHV9549, so that sucrose in molasses is converted into fructooligosaccharide. The prepared saccharide solution is treated with a Ca type strongly cacidic cation exchange resin, to give the aimed sweetener comprising fructooligosaccharide and raffinose as main components.

Description

Detailed Description of the Invention "Industrial Application Field" This invention produces a saccharide mixture containing fructooligosaccharide and raffinose as main components from sugar beet molasses. This invention relates to the production of a sweetener that promotes the proliferation of all useful bacteria in the intestines, which serve as carbon sources for bacteria.

``Prior Art'' Sugar beet molasses is a by-product during the production of sugar beet and has a unique odor, and is mainly used as feed or a raw material for fermentation, and is extremely rarely used directly as a sweetener.

On the other hand, fructooligosaccharide is disclosed in JP-A-56-154969
As described in the issue, bacteria of the genus Aureobasidium and molds of the genus Aspergillus having the ability to produce fructosyltransferase are cultured in a sucrose solution, or asparagus, Jerusalem artichoke, etc. are cultured in a sucrose solution. It is also known that Bifidobacterium has the ability to assimilate oligosaccharides obtained by catalyzing them with enzymes isolated from plants. Furthermore, it is known that roughnose exists in raw sugar beet and migrates into sugar beet molasses, and that many bifidobacteria have assimilability.

``Means and actions that solved the problem'' The inventors conducted research on sweeteners that can be used in food processing, etc. from sugar beet molasses, and found that if hooktooligosaccharides and raffinose coexist, bifidobacterium Even under the conditions where raffinose becomes a medium for fructosyltransferase and converts sea-close in sugar beet molasses into fructooligosaccharides, most of it remains due to its weak substrate specificity, and this We learned that raffinose and fructooligosaccharides flow out in almost the same fraction when chromatographically separated using a resin column, and we used the Caa-type molasses obtained in the first step of treating raffinose Kanakura and sugar beet molasses with fructosyltransferase. Combined with the second step of passing the liquid through a strongly acidic cation exchange resin column to obtain a sugar solution containing fructooligosaccharides and raffinose, when ingested by humans, it becomes a carbon source for Bifidobacterium in the intestines and promotes its growth. The problem was solved by using a sweetener.

The sugar beet molasses used in this invention is molasses obtained by a purification method such as a stephen method, a non-stephen method, or an ion exchange method during sugar production, and an example thereof is shown in Table 1.

As shown in Table 1 above, the ratio of raffinose in the six vegetable molasses varies depending on the manufacturing method, but any molasses can be used in this invention.

1. Converting the sucrose from the above molasses to fructo-oligosaccharides
IIi conversion is preferably performed using immobilized fructosyltransferase, and all known bacterial strains can be used as the fructosyltransferase source, such as Aureobasidam pullulans AHV9549.
M strain contains 20% sucrose, 1% l”JaNOs, M
g5O<・71 (20+.

0.05%, K2HPO4Q, 5% corn staple liquor, 2%, and 0.4% urea. The mixture is kneaded and added to a 10% calcium chloride solution to form granules, which are then shaped into immobilized boxes. The immobilized enzyme box produced in this manner usually has a fructosyltransferase activity of 20 to 40 units/d dry matter.

The reaction between the molasses and the immobilized enzyme can be carried out in a fixed bed system or a fluidized bed system.
L1 A jacketed column with a height of 50α was filled with 31k of the above immobilized boxed enzyme, and I-filtered molasses adjusted to pH 5 was heated to 40 to 60°C + 0 per m-body enzyme valley volume.
The liquid is passed upward from the bottom of the column at a flow rate of 2 volumes (600 cc/H). Table 2 shows the average composition of the reaction solution when the above ion exchange resin method was used for 30 consecutive days.

Table 2 However, % is the ratio to the total sugar, and GF3 and GF4 also include the transferred sugar of raffinose. Sea-crose also contains melibiose.

As is clear from Table 2, the amount of raffinose in molasses is about 7
0ti remains, and when the produced fructooligo 4i is added, 58.1% of the total sugars are occupied by oligosaccharides. Furthermore, it is also possible to return the reaction solution to the stock solution side and cause the reaction to occur again to improve the rearrangement rate.

The first step of the present invention is as described above, and in order to separate glucose, etc. from the treated sugar solution, chromatographic separation is performed using a column of Ca type strongly acidic cation exchange resin as the second step. The ion exchange resins used are Amberlite ■几-120, DOWEX 50'WX6,
A resin such as Diaion 5K-IA (both are part names) with a degree of crosslinking of 4 to 6 and a particle size of 50 to 100 mesh is particularly preferred.

The resin is treated with a solution such as fcaclz to form a Ca type, and the resin is filled into a long and narrow resin tower. Next, the treated sugar solution is passed through the tube at a temperature of 30 to 60 DEG C., and extruded with hot water at the same temperature. Now, the results of using DOWEX sOwx6 (product name) are shown in the 3rd page.
Shown in the table.

As is clear from Table 3, the sugar that first flows out is fructooligosaccharide, and raffinose also shows almost the same pattern. The next sugar that flows out is sucrose, and finally monosaccharides such as glucose.

Fractions 1 to 5 and fractions 1 to 9 in Table 3 were collected, concentrated to Bx 50, decolorized with activated carbon, and further concentrated to Bx 7.
5, the sugar mixture composition shown in Table 4 is obtained.

Table 4 As is clear from Table 4, the sugar mixture is mainly oligosaccharides, and 7-lactones 1 to 5 contain 97.4% oligo S, 1 to
9 is 90%, raffinose also accounts for about 1/4 of the total sugar, and contains a small amount of sucrose, making it a sweetener with a mild taste.

The sugar mixture in Table 4 is an artificial gastric juice (02% salt, 0 pepsin).
32% and adjusted to pH 1.5).
Even if kept at
When cultured with bifidus such as idbacterium tongum, it shows extremely good growth. From this, the sweetener of this invention reaches the intestines after being taken into the body, and is well known as a useful bacteria for humans (for example, Tomozoku Mitsuoka, "Clinical and A1 Kyō" Vol. 2, t3), p. 197 (
(1975)) serves as a carbon source for bifidus.

The sweetener of this invention may be used as it is, may be mixed with other foods, or may be mixed with a sugar having a bifidus effect such as lactulose.

Example Next, this invention will be explained in detail.

Bx sugar beet molasses using the ion exchange resin desalination method shown in Table 1
After diluting to 60% and adjusting to pH 5, the 16kg't-
201! Aureobasidum pullulans AHV9549 alginate containing 30 units/Ing dry matter of fructosyltransferase was added with calcium entrapment immobilization (500 S'e in total) and kept at that temperature for 5 hours with stirring. After the reaction was completed, the female body and the sugar solution were separated by centrifugation, and the m1 body was returned to the same tank and the enzymatic reaction was repeated 10 times.

Next, the inner diameter is 87 mm, the cylinder length is 250 mm, and the resin layer height is 170 m.
Particle size 5.0 to 100 in jacketed stainless steel column
Metsuyu's DOWEX 5 (lX4 (product name) side?l
- 1 m3 filled with the above enzyme treatment solution 1251 at a temperature of 60 ° C.
It was passed through at s v 1.3 and extruded with hot water at 60'C to obtain 9I in sugar solution 'Th230 of oligosaccharide section.

By completely lacking this sugar solution, I type H DOWEX HCR-vV2 (
Product name) 21 columns and 0) (type Revachit Ca924
9 (trade name) 4J column for desalting. Next, it was compressed to 13x50 in vacuum, decolorized by adding activated carbon, and after P was removed, it was further compressed to Bx75 to obtain 20 kg of a sugar solution with an oligo shield purity of 90%. The obtained sugar solution containing fructooligosaccharides and raffinose was a sweetener suitable for promoting the growth of bifidobacteria and had a mild sweet taste.

Effects of the Invention The present invention, as described above, involves making the sucrose contained in inexpensive sugar beet molasses contain raffinose.I. Producing fructooligosaccharide and recovering it from a mixture containing fructooligosaccharide and raffinose as main components. This method has the advantage that a sweetener that is effective for the growth of bifidobacteria can be produced at an extremely low cost and in high yield.

Claims (1)

[Claims] The first step is to treat sugar beet molasses containing raffinose with fructosyltransferase to convert sucrose into fructooligosaccharides, and the sugar solution obtained in the first step is separated using a column of Ca-type strongly acidic cation exchange resin. A method for producing a sweetener that promotes the growth of bifidobacteria from sugar beet molasses, comprising a second step of obtaining a mixture containing sugar and raffinose as main components.