JP2000041694A - Substance having high lactosucrose, its production and use - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a substance having a high lactose content and to provide a method for producing the substance and the use of the substance. SOLUTION: An aqueous solution containing sucrose and lactose is treated with a glycosyltransferase to give a lactosucrose-containing solution, from which contaminants are removed to give a solution having a high lactosucrose content of >=45 w/w% lactosucrose in a lactosucrose composition. The solution is collected as it is or spray-dried to produce a liquid or powdery substance having a high lactosucrose content. The substance having a high lactosucrose content is included to establish an oral or transluminal intake material showing selective proliferation promotion effect on bifidobacteria in vivo.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a lactosucrose-rich substance, a method for producing the same, and a use thereof. More specifically, the present invention relates to an aqueous solution containing sucrose and lactose, which is obtained by reacting an aqueous solution containing sucrose with lactose. Contaminant saccharides are removed from the lactosucrose-containing saccharide solution to obtain 45 w
/ W% or more of a lactosucrose-rich liquid, which is collected as it is or spray-dried to obtain a liquid or powdery lactosucrose-rich material, a method for producing the same, and a lactosucrose-rich material. Oral or tube ingestion.

[0002]

2. Description of the Related Art Lactosucrose is a trisaccharide consisting of galactose, glucose and fructose,
Its chemical structure is O-β-D-Gal- (1 → 4) -O
-Α-D-Glc- (1 ← 2) -β-D-Fru (however, Gal indicates a galactose residue, Glc indicates a glucose residue, and Fru indicates a fructose residue). This substance is known to be a very useful saccharide as a selective saccharide source for bifidobacteria as described in Japanese Patent Application Publication No. 53835/1984.

[0003] As a method for producing lactose, it is known that a sugar transferase such as fructose transferase or galactose transferase is allowed to act on an aqueous solution containing sucrose and lactose. Examples of the fructose transferase include, for example, Japanese Patent Application Publication No. 58905/1982 and Patent Application Publication No. 5/1979.
No. 3835, Bacill (Bacill)
rus) genus, Aerobacter
Further, levansucrase derived from a microorganism belonging to the genus Arthra belongs to the genus Arthrobacter described in Japanese Patent Application No. 160660 filed on June 26, 1999 by the present inventors. Microorganisms belonging thereto, for example, Arthrobacter sp.
obactor sp. ) K-1 (FERM PB-3)
192) can be advantageously used, and examples of galactose transferase include, for example, Canadian Journal of Chemistry.
Istry), Vol. 43, pp. 2259-2264 (1
965) and Japanese Patent Application Publication No. 85090/1988.
microorganisms belonging to the genus oboromyces) and Heisei 2
In the microorganisms belonging to the genus Rahnella described in Japanese Patent Application Publication No. 35095, and these microbial cells or extracts thereof are usefully used.

[0004] However, lactosucrose produced by allowing these glycosyltransferases to act on an aqueous solution containing sucrose and lactose usually contains only about 5 to 30% of a sugar composition. Dry as it is,
The powdered product has a drawback that it is very easy to absorb moisture, easily solidifies, and only an unstable powder that loses fluidity can be obtained.

[0005]

In utilizing lactose as a sugar source for bifidobacteria in a wide range of ingestions, a lactose-rich product containing a high concentration of lactose and a stable lactose-rich product are used. It is strongly desired to establish an industrial manufacturing method. However, the manufacturing method has not been established yet.

[0006]

Means for Solving the Problems The present inventors have intensively studied an industrial production method for obtaining a high content of lactosucrose from a sugar solution containing lactosucrose produced by a glycosyltransferase.

As a result, a glycosyltransferase is allowed to act on an aqueous solution containing sucrose and lactose, and contaminating saccharides are removed from the resulting lactosucrose-containing saccharide solution to obtain a sugar composition of 4%.
Lactosucrose-rich liquid of 5 w / w% or more was obtained, and it was found that a lactosucrose-rich material could be easily and mass-produced by collecting the solution, and in addition, lactose was contained by containing it. The present inventors have found that various oral or tube ingestible products can be easily produced, and completed the present invention.

[0008] That is, according to the present invention, first, a sugar solution is produced by producing a lactosucrose by allowing a glycosyltransferase to act on an aqueous solution containing sucrose and lactose.

At this time, the ratio of sucrose to lactose is usually selected from the range of about 1:10 to 10: 1, and the concentration thereof is generally selected from the range of about 10 to 60 w / w% in total. The pH at the time of action may be selected from a stable pH and an optimum pH of the glycosyltransferase to be used.
Is selected from the range. The temperature at the time of the action is selected from the stable temperature and the optimum temperature of the glycosyltransferase used, and is usually selected from the range of 5 to 70 ° C.

[0010] As the glycosyltransferase, the above-mentioned fructose glycosyltransferase or galactose glycosyltransferase may be allowed to act in a free state or in an immobilized state.
A sugar solution containing about 5 to 30 w / w% of lactosucrose in sugar composition is obtained.

Next, in order to increase the lactosucrose content of the present sugar solution, the present sugar solution is used as it is or according to a conventional method.
After decolorization, desalting, after concentration if necessary, a method for removing contaminating saccharides, desirably, using only water without using an organic solvent, for example, an alkali metal type or alkaline earth metal type strong acid cation One or more purification methods such as column chromatography using an exchange resin, a method for removing monosaccharides by fermentation using an invertase-deficient yeast, and a method for selectively crystallizing and removing lactose with low solubility By doing so, 45
A lactosucrose-rich liquid of w / w% or more is obtained. Examples of invertase-deficient yeast used in the fermentation method include, for example, JP-A-61-231993 and JP-A-62-23.
Yeast disclosed in Japanese Patent No. 14792 or a commercially available invertase-deficient yeast can be used. Furthermore, after lactose is selectively crystallized and removed, column chromatography using an alkali metal type or alkaline earth metal type strongly acidic cation exchange resin, one or two types selected from fermentation methods using invertase-deficient yeast The contaminating saccharides can be removed by the above method to obtain a lactosucrose-rich material.

The alkali metal type or alkaline earth metal type strongly acidic cation exchange resin used in the present invention is an alkali metal such as a Na ⁺ type or K ⁺ type styrene-divinylbenzene crosslinked copolymer having a sulfone group bonded thereto. Type, or M
One or more of alkaline earth metal types such as g ⁺⁺ type and Ca ⁺⁺ type are appropriately used. As a commercially available product, for example, Dowex 50WX manufactured by Dow Chemical Company
1, Dowex 50WX2, Dowex 50WX4
50, trade names Amberlite CG-120, Amberlite CG-6000, manufactured by Rohm and Haas
Amberlite XT-1022E and Amberlite XT-1007 manufactured by Tokyo Organic Chemical Industry Co., Ltd., and Diaion SK1 manufactured by Mitsubishi Chemical Industry Co., Ltd.
B, Diaion SK102, Diaion SK104
and so on.

More specifically, the column chromatography method is as follows. An alkali metal type or alkaline earth metal type strongly acidic cation exchange resin is suspended in water and packed in a column, and the total length of the resin layer is usually 3 m or more. Desirably, it should be 5 m or more. The temperature in this column is 30 to 7
While maintaining the temperature at 0 ° C., the concentration is about 40 to 70 w / w%
The lactosucrose-containing sugar solution as a raw material was added to the resin in an amount of about 1 to 50 v / v%, and water was flowed at an SV of about 0.1 to 2.0 at a flow rate of up or down to elute fractions. Then, the desired lactosucrose-rich fraction is collected to obtain a lactosucrose-rich liquid having a sugar composition of 45 w / w% or more.

At this time, a lactosucrose-poor fraction is collected separately from the lactosucrose-rich fraction, and a part of the eluate is replaced with the lactosucrose-poor fraction in the next chromatography. The raw sugar solution, then the lactosucrose-poor fraction, and further the elution water are flowed in that order to fractionate, so that the lactosucrose-rich liquid can be collected at a high concentration and a high yield. At this time, any of a fixed bed system, a moving bed system, and a simulated moving bed system may be used.

The lactosucrose-rich saccharified solution thus obtained is used as it is or, if necessary, decolorized, desalted, concentrated and spray-dried to obtain a lactosucrose-rich powder. . As the spray drying method, various spray drying methods such as a known rotating disk method, a pressure nozzle method, and a two-fluid nozzle method can be appropriately selected.

The sugar composition thus obtained has a composition of 45 w /
It was found that the powder having a high lactosucrose content of not less than w% was a stable powder having good fluidity and low hygroscopicity at any of the injection angle, the discharge angle and the slip angle. This powder may be used as it is or in combination with other suitable substances, in powder form or, if necessary, for example, formed into various shapes such as granules, spheres, short rods, cubes, tablets, etc. Not only as a selected sugar source, but also a sweetener, an excipient, a stabilizer, a thickener, a carbohydrate inhibitor, a humectant, a shining agent, and a mineral such as calcium, magnesium, phosphorus or a salt thereof. It has been found that it can be widely used for various oral and tube ingestions as a blended mineral compound.

In particular, the powder has an elegant sweetness,
Its sweetness is about half that of sugar, and it is in harmony with various substances having other tastes such as sourness, salty taste, astringency, umami, and bitterness. In addition, it can be advantageously used not only as a selective sugar source for Bifidobacteria but also as a sweetener, a taste improver and the like.

Further, for domestic animals, poultry, other breeding animals such as bees, silkworms, fish, etc., they can be used in feeds, feeds, pet foods and the like for the purpose of selecting a source of bifidobacteria and improving palatability.

In addition, oral medicines, tube nutrition, troches,
Bifidobacterium selective sugar source for various medicines such as liver oil drop, probiotic agent, intestinal medicine, mouth freshener, mouthwash tablet, cosmetics,
It can be advantageously used as a sweetener, a flavoring agent, an excipient, a stabilizer, a thickener and the like.

As described above, the ingestion referred to in the present invention includes a sweetener containing a high content of lactosucrose of the present invention, a mineral compounding agent, a food and drink, a favorite food, a feed, a feed, a pharmaceutical, It means all things used orally or by tube such as cosmetics.

In the present invention, the expression of a substance containing a high content of lactosucrose in the present invention means that the production of the ingested product is completed until the production of the ingested product is completed. As the method, known methods such as mixing, kneading, dissolving, dipping, spraying, coating, spraying, and pouring are appropriately selected.

When various oral or tube ingested products containing the thus obtained lactosucrose-rich material are used as a selective sugar source for bifidobacteria, one day per kg of human or animal body weight may be used. The amount of lactosucrose is usually selected from 0.01 to 5.0 g per day, and it is preferable to take this amount every day or every other day. In this case, lactosucrose is not substantially digested and absorbed to the small intestine, and bifidobacteria selectively assimilate and grow it mainly in the large intestine.

Therefore, lactosucrose has almost no increase in blood sugar level due to its ingestion, does not stimulate insulin secretion, and can be said to be a substantially low calorie or diet sweetener. In general, in order to expect a high selective growth effect of bifidobacteria in the obtained ingested food, it is preferable to contain a lactose-rich substance in an amount of about 0.1 w / w% or more, preferably 0.5 w / w% or more, as lactosucrose. It is.

[0024] The selective growth of Bifidobacteria produces organic acids such as acetic acid and lactic acid, lowers the pH in the large intestine, and suppresses the growth of harmful bacteria such as bacteria causing spontaneous infections and putrefactive bacteria. . It also suppresses the generation of harmful putrefactive substances such as ammonia, indole and cresol produced by putrefactive bacteria. In addition, it moderately stimulates the intestine, moderately promotes peristalsis, and exhibits intestinal action. In addition, the production of organic acids by Bifidobacteria promotes mineral absorption, promotes bone formation, and can be advantageously used for treatment and prevention of osteoporosis and the like.

Due to these effects, a lactosucrose-rich substance or various intakes containing the same can contribute to the maintenance and promotion of beauty and health, as well as prevent and prevent adult diseases such as diabetes, hypertension, and colorectal cancer. Medium, promotion of recovery after disease,
It can be advantageously used for treatment and prevention of hyperammonemia, hepatic encephalopathy and the like. Further, it can be advantageously used for prevention of infection of domestic animals such as livestock and poultry, prevention of diarrhea, enhancement of appetite, promotion of fattening, promotion of spawning, and suppression of odor in feces.

Hereinafter, the present invention will be clarified by experiments.

Experiment 1 Lactosucrose with glycosyltransferase
Experiment 1-1 Preparation of levansucrase defatted soybean 3 w / v%, glucose 2 w / v%, sugar
Loin 4w / v%, (NH₄)₂HPO₄ 0.6w
/ V%, MgSO₄・ 7H₂O 0.03w / v%, K
C 10.02w / v%, calcium acetate 0.02w
/ V%, MnSO ₄・ 4H₂O 0.001w / v%, water
A 60 L liquid medium consisting of tap water was adjusted to pH 7.0, and 1
After sterilizing at 20 ° C for 20 minutes, Bacillus subtilis
Squirrel (Bacillus subtilis) ATCC
6051 was inoculated and cultured at 37 ° C. for 3 days with aeration and stirring.
After completion of the culture, the culture was centrifuged to collect the supernatant, which was then collected.
Add an equal volume of cold ethanol to the mixture and centrifuge the resulting precipitate.
And then harvested containing 2 mM calcium chloride.
This solution was dissolved in 0 mM acetate buffer (pH 5.0).
Was dialyzed overnight against the same buffer, and the supernatant obtained by centrifugation was subjected to DEA.
Flow through a column filled with E-cellulose,
Adsorbed, washed with the same salt buffer, and washed with 1M NaCl.
Elute with the same acetate buffer containing
90% saturation, collect the resulting precipitate by centrifugation,
Dissolve in the same 500 mL of acetate buffer
Solution. This enzyme solution shows an activity of about 120 units / mL.
did.

The method for measuring the activity of levansucrase is as follows. A total of 2 mL reaction containing 10 w / v% sucrose, 50 mM phosphate buffer, pH 7.0 and levansucrase is kept at 30 ° C. After 30 minutes, the reaction solution is heated to inactivate the enzyme, and the amount of generated glucose is measured by a glucose oxidase method. One unit of levansucrase is defined as the amount of enzyme that produces 1 μmol of glucose per minute under these conditions.

Experiment 1-2 Preparation of Lactosucrose-Containing Sugar Solution After heating and dissolving 5 parts by weight of sucrose and 3 parts by weight of lactose in 9 parts by weight of water, cooled to 40 ° C.
The levansucrase prepared by the method of Example 1 was added at a rate of 1 unit per g of sucrose, and kept at 40 ° C. and pH 7.0.
After reacting for 4 hours, the enzyme was inactivated by heating, filtered, and the filtrate was decolorized with activated carbon according to a conventional method, then desalted with H-type and OH-type ion exchange resins, and concentrated to a concentration of about 60%.
A w / w% sugar solution was obtained. The present saccharide solution is a saccharide mixture of glucose, fructose, sucrose, lactose, lactose and the like. Of these, lactosucrose contains about 25% w / w in terms of saccharide composition. Only unstable powders with very high hygroscopicity were obtained.

Experiment 2 Influence of Lactosucrose-Containing Material on Lactosucrose Content and Its Stability The lactose-containing sugar solution used as a raw material was prepared in Experiment 1-2. The resin for fractionation was an alkali metal type strongly acidic cation exchange resin (manufactured by Tokyo Organic Chemical Industry Co., Ltd., trade name: Amberlite XT-1007, Na ⁺ type, degree of crosslinking: 6%), and a jacket having an inner diameter of 5.4 cm. A stainless steel column was filled with an aqueous suspension, and two columns were connected so that the liquid flowed in series, so that the total length of the resin layer was 10 m. While maintaining the temperature in the column at 55 ° C, the raw sugar solution was added at 5 v / v% to the resin, and hot water at 55 ° C was added to the solution.
V. 0.5 fractions were obtained to obtain various lactosucrose-rich liquids, which were decolorized, desalted, concentrated, spray-dried and powdered to a water content of less than 2 w / w% according to a conventional method. I got Lactose sucrose powder of various contents was placed in an aluminum container and left in an environment of a relative humidity of 50% and a temperature of 25 ° C. for 2 days, and the degree of moisture absorption (weight increase), fluidity, and presence or absence of consolidation were observed. . Further, in order to quantitatively grasp the degree of fluidity, various angles of repose, that is, injection angle, discharge angle, and slip angle were measured. The injection angle is 110m in diameter of the cylindrical part
m, a height of 20 mm, a height of a conical portion of 100 mm, and a polyethylene funnel having a diameter of 10 mm at the outlet are held at the level of the outlet at a height of 80 mm, and the lactosucrose powder of various contents filled in the funnel is allowed to fall naturally. The angle of the falling object was measured and measured. The discharge angle is 80 mm in diameter, 40 mm in height, and various contents of lactosucrose powder filled in a polyethylene cylindrical container having an opening with a diameter of 25 mm at the bottom are discharged from the opening at the bottom. The angle was determined by measuring. The slip angle was determined by placing 10 g of lactosucrose powder of various contents on a glass plate having a length of 95 mm, a width of 40 mm and a thickness of 5 mm and gradually tilting the glass plate, and measuring the angle at which the lactosucrose powder began to slide. The results are summarized in Table 1.

[0031]

[Table 1]

As is clear from Table 1, the sugar composition is 45 watts.
It has been found that a lactosucrose-rich content of / w% or more has low hygroscopicity, good stability, and is easy to handle.

Experiment 3 Effect of Lactosucrose High Content on Human Intestinal Flora In order to confirm that a lactosucrose high content powder is useful in vivo as a bifidobacterium selective sugar source, two healthy subjects were examined. A 47-year-old man) was orally administered with 20 g of the lactosucrose containing about 55% of the sugar composition obtained by the method of Experiment 2 every day for 10 days in 3 to 4 divided doses. The flora was searched for by weighing 1 g immediately after collecting the stool sample, placing it in 9 mL of diluent,
"World of Intestinal Bacteria (Anaerobic Separation and Fixation)", 53-6
The search was performed at the bacterial group level in accordance with the method described on page 5, Sobunbunsha (1984). The flora was searched once or twice before administration, and on days 5 and 10 during the administration period.
1 or 2 times after the administration was stopped.

The results were as follows for each of the two subjects.
This is shown as FIG. 1 and FIG. As is evident from both figures, administration of the high content of lactosucrose significantly affected the microflora, particularly causing an increase in bifidobacteria (Bifidobacterium group) and a decrease in Bacteroidase group.

As a result, the ratio of bifidobacteria to the total flora was 1.2 days in the subject of FIG.
From 0% to 70.22%, 16.96 in the subject of FIG.
Percentage has increased to 93.44%. Abdominal symptoms such as abdominal pain, diarrhea, flatulence, constipation, and discomfort did not occur during the entire test period.

Therefore, the lactosucrose-rich material of the present invention can be advantageously used as a selective sugar source for Hifidobacteria.

In the following, Example A describes a substance containing a high content of lactosucrose, and Example B describes various ingestions using the powder.

Example A-1 Lactose High Content 3 parts by weight of sucrose and 3 parts by weight of lactose were dissolved by heating in 10 parts by weight of water, cooled to 40 ° C.
-1 sucrose g
The reaction was continued for 16 hours while maintaining the temperature at 40 ° C. and the pH at 6.0. Then, the enzyme was deactivated by heating, filtered, and the filtrate was decolorized with activated carbon according to a conventional method.
The mixture was desalted with a type- and OH-type ion exchange resin, and concentrated to obtain a sugar solution having a concentration of about 50% w / w. This sugar solution has a sugar composition of about 3
It contained 0% w / w lactosucrose.

The fractionation resin used was an alkali metal-type strongly acidic cation exchange resin (manufactured by Dow Chemical Company, trade name: Dowex 50WX4, K ⁺ type, degree of crosslinking 4%), and a jacketed stainless steel having an inner diameter of 5.4 cm. The column was filled with an aqueous suspension, and four columns were connected so that the liquid flowed in series, so that the total length of the resin layer was 8 m. Column temperature 6
While maintaining the temperature at 0 ° C., the raw sugar solution is added to the resin at about 10 v / v
Then, hot water at 60 ° C. was flowed at an SV of about 0.3 to fractionate, and a lactosucrose-containing liquid having a sugar composition of about 50 w / w% was obtained. The liquid was decolorized and desalted according to a conventional method, and concentrated. And
Spray drying to obtain a powder having a water content of less than 2% w / w
%.

The lactosucrose-rich material is a powder having low hygroscopicity, good fluidity and stability, and is easy to handle.

The product can be used as such or in combination with other appropriate substances to serve not only as a selective sugar source for Bifidobacterium but also as a sweetener, excipient, thickener, etc. It can be used advantageously.

Example A-2 Lactosucrose High Content About 30 w / l lactosucrose obtained by the method of Example A-1
Except for using a w% -containing sugar solution as a fractionation resin, an alkaline earth metal type strongly acidic cation exchange resin (manufactured by Mitsubishi Kasei Kogyo Co., Ltd., trade name Diaion SK104, Ca ⁺⁺ type, degree of crosslinking 4%) Was fractionated in the same manner as in Example A-1 to obtain a liquid containing about 75% w / w of lactosucrose by sugar composition, and after decolorization and desalting, concentrated, spray-dried, and dried
Less than w / w% powder was obtained in about 30% yield.

The lactosucrose-rich material is a powder having low hygroscopicity, good fluidity and stability, and is easy to handle.

This product can be used as it is or in combination with other appropriate substances as it is in Example A-1, and can be used not only as a selective sugar source for Bifidobacteria but also as a sweetener, a stabilizer, and a shaping agent. It can be advantageously used for various oral or tube ingestions as agents, thickeners and the like.

Example A-3 Lactosucrose High Content (1) Arthrobacter sp. K-1 (FER)
MPB-3192) and cultured at 37 ° C. for 2 days. One platinum loop was taken out, and yeast extract 1.2 w / v was used.
%, Polypeptone 0.8 w / v%, soluble starch 4 w /
v%, (NH ₄ ) ₂ HPO ₄ 0.4 w / v%, MgS
_{O 4 · 7H 2 O 0.1w /} v%, liquid medium consisting of tap water (pH 7.0) to a 500mL volume shaking flask 60
The solution was dispensed in mLs and inoculated into a sterilized one, followed by shaking culture at 37 ° C. for 5 days. After completion of the culture, the culture was centrifuged, and β-
About 1.1 L of a supernatant containing fructofuranosidase was obtained.
This enzyme solution showed a β-fructofuranosidase activity of about 30 units / mL.

The method for measuring the activity of β-fructofuranosidase is as follows. 20% sucrose solution containing 40% xylose (500 mM phosphate buffer, pH 6.5) 20
Add 200 μL of the enzyme solution appropriately diluted to 0 μL, and add
After 10 minutes of action at 10 ° C., a part of the reaction solution was put into boiling water to inactivate the enzyme, and then the amounts of glucose and fructose released were measured using an F-1 kit (Boehringer Mannheim Yamanouchi Co., Ltd.) And the amount of fructose transferred is calculated by subtracting the amount of fructose from the amount of glucose. One unit is defined as the amount of enzyme that transfers 1 μmol of fructose per minute.

(2) 1 part by weight of sucrose and lactose 1
And dissolved in 3 parts by weight of water and then cooled to 50 ° C., and β-fructofuranosidase prepared by the method (1) was added thereto at a ratio of 5 units per g of sucrose.
After the reaction was carried out for 5 hours while maintaining the temperature at 6.5 ° C. and the pH 6.5, the enzyme was inactivated, filtered, and the filtrate was decolorized and desalted according to a conventional method.
It was concentrated to obtain a sugar solution having a concentration of about 60 w / w%. This sugar solution is
The sugar composition contained about 30% w / w lactosucrose.

The present sugar solution was used as a fractioning resin in Example A-
Fractionation was performed in the same manner as in Example A-1, except that the resin used in Step 2 was changed to an alkaline earth metal type (Mg ⁺⁺ type), and a liquid containing about 55 w / w% of lactosucrose in sugar composition was obtained. Similarly, purification, concentration, and spray drying were performed to obtain a powder having a water content of less than 2% w / w in a yield of about 45%.

The lactosucrose-rich powder is a powder having low hygroscopicity, good fluidity and stability, and easy to handle.

This product can be used as it is or in combination with other appropriate substances as in Example A-1 to provide not only a selective sugar source for Bifidobacteria, but also sweeteners, stabilizers, and excipients. It can be advantageously used for various oral or tube ingestions as agents, thickeners and the like.

Example A-4 Lactosucrose High Content About 30 w / lactose of lactosucrose obtained by the method of Example A-3
w% -containing sugar solution is used as a fractionation resin as an alkali metal type strongly acidic cation exchange resin (manufactured by Rohm and Haas Co., trade name Amberlite CG-6000, Na ⁺ type,
Except for using a crosslinkage degree of 6%), fractionation was carried out in the same manner as in Example A-1 to obtain a liquid containing about 70% w / w% of lactosucrose in sugar composition, and further purified, concentrated and spray-dried in the same manner. , A powder having a water content of less than 2 w / w% was obtained in a yield of about 35%.

The lactosucrose-rich powder is a powder having low hygroscopicity, good fluidity and stability, and is easy to handle.

This product can be used as it is or in combination with other appropriate substances as it is in Example A-1, and can be used not only as a selective sugar source for bifidobacteria but also as a sweetener, a stabilizer, and a vehicle. It can be advantageously used for various oral or tube ingestions as agents, thickeners and the like.

Example A-5 Lactosucrose-rich material 1 part by weight of sucrose and 1 part by weight of lactose were dissolved by heating in 2 parts by weight of water, and then cooled to 50 ° C.
-3. The β-fructofuranosidase prepared by the method of (1) was added at a ratio of 8 units per g of sucrose.
The reaction was maintained at H 6.0 for 20 hours, then the enzyme was deactivated and filtered to obtain a filtrate.

[0055] This solution contained monosaccharides such as glucose and fructose, and oligosaccharides such as lactose and sucrose as contaminating sugars in addition to about 35 w / w% of lactosucrose in sugar composition.

This solution was adjusted to a sugar concentration of about 30 w / w%, and an invertase-deficient yeast (trade name “LS-Y”, manufactured by Oriental Yeast Co., Ltd., water content: about 65%) was added at 5 w / w per solid sugar. %, PH 6.5 to 7.0, temperature 3
The mixture was kept at 0 ° C. for 24 hours, and the monosaccharides contained in the sugar solution were fermented and digested to give a solution containing about 46% w / w of lactosucrose in the sugar composition, and then decolorized, desalted, concentrated and concentrated according to a conventional method. Spray-dry to reduce powder with less than 3% moisture to about 64% yield
I got it.

The lactosucrose-rich material is a powder having low hygroscopicity, good fluidity and stability, and is easy to handle.

This product can be used as it is or in combination with other appropriate substances as it is in Example A-1, and can be used not only as a selective sugar source for bifidobacteria, but also as a sweetener, a stabilizer, and an excipient. It can be advantageously used for various oral or tube ingestions as agents, thickeners and the like.

Example A-6 Lactosucrose High Content About 35 w / lactose of lactosucrose obtained by the method of Example A-5
w% -containing sugar solution is concentrated to a concentration of about 78 w / w%,
Then, a small amount of crystalline lactose was added as a seed crystal, and the lactose was crystallized while gradually cooling to 30 ° C and stirring.
The crystalline lactose is removed by centrifugation and filtration to give a liquid containing about 46% w / w of lactosucrose in sugar composition, then decolorized, desalted, concentrated, spray-dried, and dried to give a water content of 3% in accordance with a conventional method. % Powder was obtained with a yield of about 66%.

The lactosucrose-rich material is a powder having low hygroscopicity, good fluidity and stability, and is easy to handle.

This product can be used as it is or in combination with other appropriate substances as in the case of Example A-1, and can be used not only as a sugar source for selecting bifidobacteria, but also as a sweetener, a stabilizer, and an excipient. It can be advantageously used for various oral or tube ingestions as agents, thickeners and the like.

Example A-7 Lactose-rich content 4 parts by weight of sucrose and 5 parts by weight of lactose were added to water.
After heating and dissolving in 5 parts by weight, the mixture was cooled to 55 ° C., and β-fructofuranosidase prepared by the method of Example A-3 (1) was added thereto at a ratio of 10 units per g of sucrose.
Reaction was carried out for 15 hours at 5 ° C. and pH 6.5, and then
The enzyme was inactivated and filtered to obtain a filtrate.

This solution contained, in addition to lactosucrose of about 33% w / w in sugar composition, monosaccharides such as glucose and fructose and oligosaccharides such as lactose and sucrose as contaminating saccharides.

This solution was concentrated to about 78% w / w, a small amount of crystalline lactose was added as a seed crystal, and lactose was crystallized while stirring at 50 ° C., and then filtered to remove crystalline lactose. The obtained filtrate was adjusted to a concentration of about 30 w / w%, and monosaccharides were fermented and digested with an invertase-deficient yeast in the same manner as in Example A-5 to obtain a liquid containing about 50 w / w% of lactosucrose in sugar composition. Then, according to a conventional method, decolorization, desalting, concentration and spray drying were performed to obtain a powder having a water content of less than 2% in a yield of about 52%.

The lactosucrose-rich material is a powder having low hygroscopicity and good fluidity and stability, and is easy to handle.

This product can be used as it is or in combination with other appropriate substances as in the case of Example A-1, and can be used not only as a selective sugar source for bifidobacteria but also as a sweetener, a stabilizer, and a shaping agent. It can be advantageously used for various oral or tube ingestions as agents, thickeners and the like.

Example B-1 Mixed Sweetener High Lactosucrose Content 1 Obtained by the Method of Example A-2
0.05 parts by weight of α-glycosyl stevioside (manufactured by Toyo Seishi Co., Ltd., trade name α-G suite) was uniformly mixed with the parts by weight to prepare a powdered sweetener. The product has an elegant sweetness, about twice the sweetness of sugar. In addition, this product has the effect of promoting the growth of bifidobacteria, and is useful for the maintenance and promotion of beauty, health, prevention of adult diseases, promotion of recovery during and after illness, treatment and prevention of hyperammonemia, hepatic encephalopathy, etc. It can be used advantageously. In addition, it can be advantageously used for prevention of infection of domestic animals such as livestock and poultry, prevention of diarrhea, enhancement of appetite, promotion of fattening, and suppression of odor in feces.

Example B-2 Hard candy 15 parts by weight of the high content of lactosucrose obtained by the method of Example A-1 was dissolved in 95 parts by weight of reduced maltose starch syrup (water 25 w / w) under heat and reduced pressure. The mixture was concentrated until the water content was less than 2 w / w%, and 1 part by weight of citric acid and an appropriate amount of a lemon flavor and a coloring agent were mixed with the concentrate, and then molded according to a conventional method to produce a hard candy.

This product is a low caries hard candy having an elegant sweetness. In addition, this product exhibits a bifidobacterium growth promotion effect, and as in Example B-1,
It can be advantageously used for maintenance and promotion of beauty and health, prevention of adult diseases, promotion of recovery during and after illness, and treatment and prevention of various diseases.

Example B-3 Chewing gum 2 parts by weight of a gum base were heated and melted to such an extent that they became soft,
After mixing 4 parts by weight of the lactosucrose-rich material obtained by the method of Example A-3, 3 parts by weight of glucose, and an appropriate amount of peppermint flavor and coloring agent, kneading with a roll according to a conventional method and molding. Produced chewing gum.

This product has good texture and sweetness. In addition, this product exhibits a bifidobacterium growth promotion effect, and similarly to the case of Example B-1, beauty and health maintenance and promotion, prevention of adult diseases, promotion of recovery during and after disease, treatment of various diseases It can be advantageously used for prevention and the like.

Example B-4 40 parts by weight of chocolate cocoa paste, 10 parts by weight of cocoa butter,
Lactosucrose-rich material 1 obtained by the method of Example A-4
5 parts by weight, 10 parts by weight of sucrose and 15 parts by weight of whole milk powder were mixed and passed through a refiner. After reducing the particle size, the mixture was placed in a conche, to which 0.5 part by weight of lecithin was added, and kneaded at 50 ° C. for two days and nights. Subsequently, it was poured into a molding machine according to a conventional method, and was molded and solidified to be manufactured.

This product has no fear of fat bloom and sugar bloom and has good melting and flavor when placed on the tongue. In addition, the product exhibits a bifidobacterium growth promotion effect, and can be used for promoting beauty, maintaining and promoting health, and promoting recovery during and after disease as in Example B-1.

Example B-5 Milk Beverage 70 parts by weight of coffee leachate, 110 parts by weight of milk, 14 parts by weight of the high-lactosucrose content obtained by the method of Example A-5, and appropriate amounts of coffee flavor and coloring agent were mixed. After homogenization, sterilization, cooling, filling, and packaging were performed according to a conventional method to produce a milk beverage.

The product is coffee milk having good aroma and taste. In addition, this product exhibits a bifidobacterium growth promotion effect, and is advantageously used for beauty, maintenance and promotion of health, prevention of adult diseases, promotion of recovery during and after illness, etc. as in Example B-1. it can.

Example B-6 500 parts by weight of custard cream corn starch, 500 parts by weight of a high content of lactosucrose obtained by the method of Example A-6, and maltose (available from Hayashibara Shoji Co., Ltd., registered trademark: San Mart) 400
Parts by weight and 5 parts by weight of salt were thoroughly mixed through a sieve, 1400 parts by weight of eggs were added, and the mixture was stirred. 5000 parts by weight of boiled milk was gradually added to the mixture. The custard cream was prepared by turning off the fire when it was completely gelatinized and becoming totally translucent, cooling it and adding an appropriate amount of vanilla flavoring.

The product has a smooth luster, is not too sweet, and is delicious. In addition, this product exhibits a bifidobacterium growth promotion effect, and as in the case of Example B-1, cosmetics,
It can be advantageously used for maintaining and improving health, preventing adult diseases, promoting recovery during and after illness, and the like.

Example B-7 Instant corn potage soup 30 parts by weight of α-modified corn powder, 5 parts by weight of α-modified starch, α
4 parts by weight of modified potato starch, 12 parts by weight of α-modified waxy corn starch, 8 parts by weight of a high content of lactosucrose obtained by the method of Example A-7, 5 parts by weight of sodium glutamate
Parts by weight, 8.5 parts by weight of salt, 7 parts by weight of skim milk powder, and 0.5 part by weight of onion powder are ground and mixed well, and then 0.5 part by weight of sorbitan fatty acid ester and 9 parts by weight of vegetable hardened oil are mixed. And heat-melted parts are added and mixed,
Further, lactose (10 parts by weight) was added and mixed, and the mixture was charged into a fluidized-bed granulator, sprayed with a small amount of water and granulated.
It was dried with hot air at 0 ° C. and sieved to produce an instant corn potage soup.

This product, when poured into boiling water, easily dissolves and disperses into a soup with excellent flavor. In addition, this product exhibits a bifidobacterium growth promotion effect, and is advantageously used for beauty, maintenance and promotion of health, prevention of adult diseases, promotion of recovery during and after illness, etc. as in Example B-1. it can.

Example B-8 Tube-feeding nutrient Lactosucrose-rich material 5 obtained by the method of Example A-1
80 parts by weight, dried egg yolk 190 parts by weight, skim milk powder 209 parts by weight, sodium chloride 4.4 parts by weight, potassium chloride 1.
85 parts by weight, magnesium sulfate 4 parts by weight, thiamine 0.
01 parts by weight, 0.1 part by weight of sodium ascorbate,
A formulation is prepared consisting of 0.6 parts by weight of vitamin E acetate and 0.04 parts by weight of nicotinamide. Each 25 g of this compound is filled into a laminated aluminum pouch,
Heat sealing was performed to produce a solid tube nutritional supplement.

This product maintains its quality for a long time,
Good solubility and dispersibility. This drug can be used for about 15 bags.
It is dissolved in 0 to 300 mL of water to prepare a nutritional supplement, and administered to the nasal cavity, esophagus, stomach and the like by the tube method.
In addition, this product exhibits a bifidobacterium growth promotion effect, and similarly to the case of Example B-1, beauty and health maintenance and promotion, prevention of adult diseases, promotion of recovery during and after disease, treatment of various diseases ,
It can be advantageously used for prevention, further promotion of recovery during and after disease of livestock and poultry, and promotion of fattening.

Example B-9 Tablets Lactosucrose-rich material 4 obtained by the method of Example A-2
After uniformly mixing 0 parts by weight, 10 parts by weight of maltose, 1 part by weight of tribasic calcium phosphate, 1 part by weight of sugar ester and an appropriate amount of powdered flavor, about 35 tablets per tablet according to a conventional method.
Tableting was performed with a tableting machine so as to obtain 0 mg to obtain tablets.

This product is an easy-to-drink tablet having good stability without cracking. By taking about 1 to 40 tablets, preferably about 2 to 20 tablets per day for an adult, the effect of promoting the growth of bifidobacteria and the absorption of calcium can be obtained. It exerts an accelerating effect, as in the case of Example B-1, as well as beauty, maintenance and promotion of health, prevention of adult diseases, promotion of recovery during and after illness, treatment of various diseases,
It can be used advantageously for prevention.

Example B-10 Formulated feed 40 parts by weight of powdered flour, 38 parts by weight of skim milk powder, 12 parts by weight of a lactosucrose-rich material obtained by the method of Example A-3, 10 parts by weight of vitamin preparation, 5 parts by weight of fish meal , 5 parts by weight of dibasic calcium phosphate, 3 parts by weight of liquid oil and fat, 3 parts by weight of calcium carbonate, 2 parts by weight of sodium chloride and 2 parts by weight of a mineral agent to prepare a compound feed.

The product is a feed for livestock and poultry with improved palatability, and is particularly suitable as a feed for piglets. In addition, this product exhibits a bifidobacterium growth-promoting effect and a mineral-promoting effect, and can be advantageously used to prevent infection of breeding animals, prevent diarrhea, increase appetite, promote fattening, and control odor in feces. In addition, the product may be used in combination with other feed materials, such as cereals, flour, starch, oil cakes, sucrose, etc., and rough feed materials such as straw, hay, bagasse, corn cob, etc., if necessary. Then, it can be made into other compound feed.

[0086]

As described in the text, the present invention establishes a high content of lactosucrose, which has been impossible so far, and an industrial production method and use thereof. That is, contaminating saccharides are removed from a lactosucrose-containing sugar solution produced by reacting a glycosyltransferase with an aqueous solution containing sucrose and lactose to give a lactosucrose-rich solution having a sugar composition of 45 w / w% or more. To establish a stable high lactosucrose content and an industrial production method thereof, and to establish an oral or tube ingestion containing the high lactosucrose content. is there.

The lactosucrose-rich material thus obtained can be used not only as a selective sugar source for bifidobacteria,
It can be widely used as a sweetener, a mineral absorption enhancer, an excipient, a stabilizer, a thickener, a carbohydrate crystal inhibitor, a humectant, an illuminant and the like for various oral or tube feedings.

Further, since the lactosucrose-rich material or various ingested foods containing the same can exert the effect of promoting the growth of bifidobacteria, it can not only contribute to the maintenance and promotion of beauty and health, but also prevent adult diseases, It can be advantageously used for promoting recovery during and after illness, treating and preventing hyperammonemia, hepatic encephalopathy and the like.

Further, it can be advantageously used for prevention of infection of domestic animals, poultry, and other domesticated animals, prevention of diarrhea, enhancement of appetite, promotion of fattening, promotion of spawning, suppression of odor in feces, and the like. The industrial significance in this field is extremely large.

[Brief description of the drawings]

FIG. 1 is a diagram showing search results of intestinal flora.

FIG. 2 is a diagram showing search results of intestinal flora.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI theme coat ゛ (Reference) A23L 1/30 A23L 1/30 Z 2/60 3/46 3/46 A61K 31/00 601 A61K 31/00 601 31/70 31/70 47/26 L 47/26 BJ C12N 9/10 C12N 9/10 A23C 9/13 // A23C 9/13 A23K 1/16 303D A23K 1/16 303 C07H 3/04 C07H 3 / 04 A23L 2/00 C (72) Inventor Kozo Hara 2-6-3-104 Namiki, Kanazawa-ku, Yokohama-shi, Kanagawa Prefecture (72) Inventor Takaki Fujita 2-6-308 Morinomiya, Joto-ku, Osaka-shi, Osaka ( 72) Inventor Masayuki Yamashita 10-16, Tsutsujigaoka 2-chome, Tarumizu-ku, Kobe City, Hyogo Prefecture Gunze Town Ejiri Asahigaoka 1-chome No. 3 No. 41 (72) inventor Toshio Miyake Okayama Ishima-cho 1-chome No. 3 No. 23

Claims (8)

[Claims] 1. A sugar transferase is reacted with an aqueous solution containing sucrose and lactose to remove contaminating saccharides from the obtained lactosucrose-containing saccharide solution to obtain 45 w / w saccharide composition.
% Or more of a liquid or powdery lactosucrose-rich liquid obtained by collecting or spray-drying a liquid having a high lactosucrose content of at least 10%. 2. An aqueous solution containing sucrose and lactose is reacted with a glycosyltransferase, and the resulting lactosucrose-containing saccharide solution is subjected to column chromatography using an alkali metal or alkaline earth metal type strongly acidic cation exchange resin, invertase. The contaminating saccharide is removed by one or more methods selected from a fermentation method using a deficient yeast and a method of crystallizing and removing lactose.
A liquid or powdery lactosucrose-rich substance obtained by collecting or spray-drying a lactosucrose-rich liquid of 5 w / w% or more. 3. An aqueous solution containing sucrose and lactose is reacted with a glycosyltransferase to remove contaminating saccharides from the resulting lactosucrose-containing saccharide solution to obtain a 45% w / w saccharide composition.
% Of a lactosucrose-rich liquid, and collecting or spray-drying the liquid as it is, or a liquid or powdery lactosucrose-rich material. 4. An aqueous solution containing sucrose and lactose is reacted with a glycosyltransferase, and the resulting lactosucrose-containing sugar solution is subjected to column chromatography using an alkali metal or alkaline earth metal type strongly acidic cation exchange resin, invertase. The contaminating saccharide is removed by one or more methods selected from a fermentation method using a deficient yeast and a method of crystallizing and removing lactose to obtain a sugar composition of 4%.
A method for producing a liquid or powdery lactosucrose-rich material, characterized in that a lactosucrose-rich liquid of 5 w / w% or more is collected and directly collected or spray-dried. 5. An oral or tube ingestion containing the lactosucrose-rich substance according to claim 1 or 2. 6. The oral or tube feed according to claim 5, wherein the lactosucrose suppresses the generation of putrefactive substances produced by putrefactive bacteria in the intestine. 7. The oral or tube ingestible product according to claim 5, which contains a mineral together with lactosucrose. 8. The oral or tube ingestion according to claim 7, wherein the mineral is calcium, magnesium, phosphorus or a salt thereof.