JPH1057092A - Substance containing lactosucrose in high content and its production and use - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable easy and mass production of the subject substance- containing solution by allowing a glycosyltransferase to act on an aqueous solution containing sucrose and lactose and removing contaminant saccharides from the resultant lactosucrose containing aqueous solution. SOLUTION: A glycosyltansferase (for example, fructose glycosy-transferase) is allowed to act on an aqueous solution containing sucrose and fructose. Then, the contaminants are removed from the resultant lactosucrose-containing saccharide solution, preferably by alkali or alkaline earth metal type storing acid cationic ion-exchange resin column chromatography using only water to give a solution containing more than 45w/w% of lactosucrose and the solution is collected. usually, the glysyltransferase is allowed to act on an aqueous solution containing sucrose and lactose at a ratio of 1/10-10/1 in the total saccharide concentration of 10-60wt.% at a pH of 3-9 and a temperature of 5-70 deg.C.

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. Contaminant saccharides are removed from the lactosucrose-containing saccharide solution to obtain 45 w
The present invention relates to a lactosucrose-rich liquid obtained by extracting a lactosucrose-rich liquid of at least / w%, a method for producing the lactosucrose-rich substance, and a method for producing the lactosucrose-rich liquid.

[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 of a 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. Commercial products include, for example, Dowex 50WX1, trade name manufactured by Dow Chemical Company,
Dowex 50WX2, Dowex 50WX45
0, Amberlite CG-120 and Amberlite CG-6000 manufactured by Rohm and Haas Company, Amberlite X manufactured by Tokyo Organic Chemical Industry Co., Ltd.
T-1022E, Amberlite XT-1007, trade name Diaion SK1B manufactured by Mitsubishi Chemical Industry Co., Ltd.,
There are Diaion SK102 and Diaion SK104.

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.

[0027]

[Experiment 1] <Preparation of lactosucrose-containing sugar solution using glycosyltransferase>

[Experiment 1-1] <Preparation of Levansucrase> 3% w / v defatted soybean, 2% w / v% glucose, 4% w / v sucrose, (N
H ₄ ) ₂ HPO ₄ 0.6 w / v%, MgSO ₄ .7H ₂ O
0.03w / v%, KC 10.02w / v%, calcium acetate _{0.02w / v%, MnSO 4 ·} 4H 2 O
A 60 L liquid medium consisting of 0.001 w / v% tap water was adjusted to pH 7.0, sterilized at 120 ° C. for 20 minutes, and then Bacillus subtilis (Bacillus su).
btilis) ATCC6051 and inoculated at 37 ° C. for 3 hours.
The culture was performed with aeration and stirring for days. After completion of the culture, the culture was centrifuged to collect the supernatant, to which an equal volume of cold ethanol was added, the resulting precipitate was collected by centrifugation, and then a 20 mM acetate buffer containing 1 mM calcium chloride was collected. (PH
5.0), and this solution was dialyzed overnight against the same buffer, and the supernatant obtained by centrifugation was passed through a column filled with DEAE-cellulose to adsorb levansucrase, washed with the same salt buffer, and washed with 1M NaCl. And eluted with 90% saturation of ammonium sulfate. The resulting precipitate was collected by centrifugation, and the same acetate buffer was used.
It was dissolved in 0 mL to obtain a levansucrase solution. This enzyme solution showed an activity of about 120 units / mL.

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.

[0029]

[Experiment 1-2] <Preparation of lactose-containing sugar solution> 5 parts by weight of sucrose and 3 parts by weight of lactose were dissolved by heating in 9 parts by weight of water, and then cooled to 40 ° C. After adding levansucrase prepared in the above at a rate of 1 unit per g of sucrose and reacting at 40 ° C. and pH 7.0 for 24 hours, the enzyme was inactivated by heating, filtered, and the filtrate was filtered with activated carbon in a usual manner. Decolorized, then desalted with H-type and OH-type ion exchange resins, and concentrated to obtain a sugar solution having a concentration of about 60 w / w%. 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.

[0030]

[Experiment 2] <Influence of Lactosucrose-Containing Material on Lactosucrose Content and Its Stability> The lactosucrose-containing sugar solution used as a raw material was prepared in Experiment 1-2. The fractionation resin is an alkali metal type strongly acidic cation exchange resin (manufactured by Tokyo Organic Chemical Industry Co., Ltd., trade name: Amberlite XT-1)
007, Na ⁺ type, degree of crosslinking 6%) and an inner diameter of 5.4
cm of a jacketed 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. Column temperature is 55 ° C
The raw sugar solution was added at 5 v / v% to the resin while maintaining the temperature at 55 ° C., and hot water at 55 ° C. was flowed at SV 0.5 to fractionate the solution.
A lactosucrose-rich liquid of various contents was obtained, and this was decolorized, desalted, concentrated, spray-dried according to a conventional method,
A powder having a water content of less than 2% w / w was obtained. Lactose sucrose powder of various contents is put in an aluminum container, and the relative humidity is 50%.
It was left in an environment at a temperature of 25 ° C. for 2 days, and the degree of moisture absorption (weight increase), the possibility of fluidity, and the 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 maintained at a height of 80 mm at the level of the outlet by holding a polyethylene funnel having a diameter of 110 mm in the cylindrical portion, a height of 20 mm, a height of the conical portion of 100 mm, and a diameter of the outlet of 10 mm. The angle of the fallen object when the lactosucrose powder was allowed to fall naturally was measured and determined.
The discharge angle is 80mm in diameter, 40mm in height and 2mm in diameter at the bottom.
The angle of the remaining lactosucrose powder formed after the lactosucrose powder of various contents filled in the polyethylene cylindrical container having the opening of 5 mm was discharged from the opening at the bottom was measured. The slip angle is 95mm in length and 40 in width
10 g of lactosucrose powder of various contents was placed on a glass plate having a thickness of 5 mm and a thickness of 5 mm, and gradually tilted, and the angle at which the lactosucrose powder began to slide was measured and determined.
The results are tabulated.

[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.

[0033]

[Experiment 3] <Effect of lactosucrose-rich substance on human intestinal flora>
To confirm its usefulness as a selective sugar source for bifidobacteria, two healthy subjects (both 47-year-old males) were
Lactosucrose about 55% in sugar composition obtained by the method of Experiment 2
The content was orally administered at 20 g per day in 3 to 4 divided doses for 10 consecutive days. To search for the microflora, 1 g of a fecal sample was immediately collected and weighed and placed in 9 mL of a diluent, and written by Tomoari Mitsuoka, “World of Intestinal Bacteria (Anaerobic Separation and Fixation)”, pp. 53-65, Monososha (1984), a search was performed at the bacterial group level. 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.

[0038]

Example A-1 <Lactosucrose high content> After dissolving 3 parts by weight of sucrose and 3 parts by weight of lactose in 10 parts by weight of water,
After cooling to 40 ° C, levansucrase prepared by the method of Experiment 1-1 was added thereto at a ratio of 1.5 units per sucrose, and the mixture was reacted at 40 ° C and pH 6.0 for 16 hours. The mixture was deactivated 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 obtain a sugar solution having a concentration of about 50 w / w%. This sugar solution contained about 30% w / w lactosucrose in sugar composition.

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 0 ° C., the raw sugar liquid is added to
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.

[0042]

Example A-2 <Lactosucrose High Content> The sugar solution containing about 30 w / w% lactosucrose obtained by the method of Example A-1 was used as a fractionation resin as an alkaline earth metal-type strongly acidic cation. Exchange resin (manufactured by Mitsubishi Chemical Industries, Ltd., trade name Diaion S
K104, Ca ⁺⁺ type, degree of crosslinking 4%)
Fractionation was carried out in the same manner as in Example A-1 to obtain a liquid containing about 75 w / w% of lactosucrose in sugar composition, and after decolorization and desalting, concentrated and spray-dried to obtain a powder having a water content of less than 2 w / w%. Was obtained in a yield of about 30%.

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.

[0045]

Example A-3 <Lactosucrose High Content> (1) Arthrobacter sp. K-1 (FER) on a normal agar slant medium
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%, MgSO _{4
· 7H 2 O 0.1w / v%} , 60mL liquid medium (pH 7.0) to a 500mL volume shaking flask made of tap water
Each was dispensed and sterilized, and inoculated, and cultured with shaking at 37 ° C. for 5 days. After completion of the culture, the culture was centrifuged to obtain about 1.1 L of a supernatant containing β-fructofuranosidase. 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-
The fractionation was carried out 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) to obtain a liquid containing about 55 w / w% of lactosucrose in sugar composition. Further, purification, concentration, and spray drying were performed in the same manner 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.

[0051]

Example A-4 <Lactosucrose High Content> An alkali metal-type strongly acidic cation exchange resin containing about 30 w / w% lactosucrose-containing sugar solution obtained by the method of Example A-3 as a fractionation resin. (Manufactured by Rohm and Haas Co., trade name Amberlite CG-6000, Na ⁺ type, degree of cross-linking 6%), and fractionated in the same manner as in Example A-1. A liquid containing 70 w / w% was obtained, and further purified, concentrated and spray-dried in the same manner to obtain a powder having a water content of less than 2 w / w% 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.

[0054]

Example A-5 <Lactose-rich sucrose> 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 dissolved in 5 parts by weight.
After cooling to 0 ° C., β-fructofuranosidase prepared by the method of Example A-3 (1) was added thereto at a rate of 8 / g sucrose.
The reaction was carried out for 20 hours while maintaining the temperature at 50 ° C. and the pH at 6.0, and then the enzyme was inactivated 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.

[0059]

Example A-6 <Lactosucrose high content> The sugar solution containing about 35 w / w% of lactosucrose obtained by the method of Example A-5 was concentrated to a concentration of about 78 w / w%, and at 50 ° C, A small amount of crystalline lactose was added as a seed crystal, and the lactose was crystallized while being slowly cooled to 30 ° C. and stirred, and then centrifuged and filtered to remove crystalline lactose, and the sugar composition was about 46% w / w of lactose in sugar composition. The resulting solution was decolorized, desalted, concentrated and spray-dried according to a conventional method to obtain a powder having a water content of less than 3% in 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.

[0062]

Example A-7 <Lactose-rich sucrose> 4 parts by weight of sucrose and 5 parts by weight of lactose were dissolved by heating in 13.5 parts by weight of water, cooled to 55 ° C., and cooled to 55 ° C. Β-fructofuranosidase prepared by the method of (1) was added to sucrose g
The reaction was carried out for 15 hours while maintaining the temperature at 55 ° C. and the pH at 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.

[0067]

Example B-1 <Mixed sweetener> α-glycosyl stevioside (manufactured by Toyo Seika Co., Ltd., trade name α-G suite) is added to 1 part by weight of the high-lactosucrose content obtained by the method of Example A-2. .
05 parts by weight were uniformly mixed to produce a powdered sweetener. The product has an elegant sweetness, about twice the sweetness of sugar. In addition, this product exerts the effect of promoting the growth of bifidobacteria, maintaining and improving the beauty and health, preventing adult diseases, promoting recovery during and after illness, treating hyperammonemia, hepatic encephalopathy,
It can be used advantageously for prevention. In addition, prevention of infection of domestic animals such as livestock and poultry, prevention of diarrhea, promotion of appetite, promotion of fattening,
It can also be used advantageously to control the odor of feces.

[0068]

[Example B-2] <Hard candy> Reduced maltose syrup (water 25w /
w) 15 parts by weight of the lactosucrose-rich material obtained by the method of Example A-1 was dissolved by heating in 95 parts by weight under heat and concentrated under reduced pressure until the water content was less than 2 w / w%, and citric acid was added thereto. Mix 1 part by weight and appropriate amount of lemon flavor and colorant,
Then, molding was performed according to a conventional method to produce 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.

[0070]

<Example B-3><Chewinggum> 2 parts by weight of a gum base were heated and melted so as to be soft, and 4 parts by weight of a lactosucrose-rich material obtained by the method of Example A-3, 3 parts by weight of glucose and an appropriate amount After mixing the peppermint perfume and the colorant, the mixture was kneaded with a roll according to a conventional method and molded to produce a 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.

[0072]

<Example B-4><Chocolate> 40 parts by weight of cocoa paste, 10 parts by weight of cocoa butter, 15 parts by weight of high-lactosucrose content obtained by the method of Example A-4, 10 parts by weight of sucrose, and whole milk powder 15 The parts by weight 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.

[0074]

<Example B-5><Milkdrink> 70 parts by weight of a coffee infusion, 110 parts by weight of milk, 14 parts by weight of a high content of lactosucrose obtained by the method of Example A-5, and an appropriate amount of coffee flavor and coloring agent After homogenization, sterilization, cooling, filling,
The milk beverage was manufactured by packaging.

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.

[0076]

Example B-6 <Custard cream> 500 parts by weight of corn starch
Lactosucrose-rich material 5 obtained by the method of Example A-6
00 parts by weight, 400 parts by weight of maltose (available from Hayashibara Shoji Co., Ltd., registered trademark of San Mart) and 5 parts by weight of salt,
Thoroughly mix through a sieve, add 1400 parts by weight of chicken eggs and stir, slowly add 5,000 parts by weight of boiled milk, further stir the mixture over a low heat, and corn starch is completely gelatinized and the whole is half in total. The custard cream was prepared by stopping the fire when it became clear, cooling it and adding an appropriate amount of vanilla flavor.

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.

[0078]

Example B-7 <Instant corn potage soup> α-modified corn powder 30
Parts by weight, α-modified starch 5 parts by weight, α-modified potato starch 4
Parts by weight, 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, 8.5 parts by weight of sodium chloride, 7 parts by weight of skim milk powder, After 0.5 part by weight of onion powder is ground and mixed well, a mixture obtained by heating and melting 0.5 part by weight of a sorbitan fatty acid ester and 9 parts by weight of a vegetable hardened oil is added thereto and mixed. 10 parts by weight of lactose was added and mixed, charged into a fluidized bed granulator, sprayed with a small amount of water and granulated, and then dried with hot air at 70 ° C.
Immediate corn potage soup was produced by sieving.

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.

[0080]

<Example B-8><Tubenutrition> 580 parts by weight of a high lactosucrose content obtained by the method of Example A-1, 190 parts by weight of dried egg yolk, 209 parts by weight of skim milk powder, 4.4 parts by weight of sodium chloride Parts, 1.85 parts by weight of potassium chloride, 4 parts by weight of magnesium sulfate, 0.01 parts by weight of thiamine, 0.1 parts by weight of sodium ascorbate, 0.6 parts by weight of vitamin E acetate and 0.04 parts by weight of nicotinamide. A formulation is prepared. Each 25 g of the blend was filled into a laminated aluminum pouch and heat-sealed to produce a solid tube-feeding nutrient.

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.

[0082]

Example B-9 <Tablet> 40 parts by weight of lactose rich content obtained by the method of Example A-2, 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 powder After uniformly mixing the fragrance, 1
The tablets were tableted with a tableting machine so as to be about 350 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.

[0084]

<Example B-10><Formulatedfeed> 40 parts by weight of powdered wheat glue, 38 parts by weight of skim milk powder, lactose-rich content 12 obtained by the method of Example A-3
Parts by weight, 10 parts by weight of vitamin preparation, 5 parts by weight of fish meal, 5 parts by weight of dicalcium phosphate, 3 parts by weight of liquid oil and fat, 3 parts by weight of calcium carbonate, 2 parts by weight of salt and 2 parts by weight of mineral preparation Was manufactured.

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.

──────────────────────────────────────────────────の Continuation of the front page (51) Int.Cl. ⁶ Identification code Agency reference number FI Technical display A61K 31/70 ACJ A61K 33/42 ABU 33/06 ADF ABY 33/42 ABU ADP ABY ADT ADP ADU ADT AEX ADU AFD AEX AGZ AFD 47/26 ABJJ AGZ ACRL 47/26 ABJ ADDB ACR C12S 3/02 ADD A23K 1/16 303D C12S 3/02 C07H 3/04 // A23K 1/16 303 A23L 3/00 C0 / 04 F (C12S 3/02 C12R 1: 865) (72) Inventor Takaki Fujita 2-6308 Morinomiya, Joto-ku, Osaka City, Osaka (72) Inventor Masayuki Yamashita 2--16 Tsutsujigaoka, Tarumizu-ku, Kobe City, Hyogo Prefecture Address 10 (72) Inventor Yasuhiko Tsunetomi Kobe, Hyogo Kita-ku Dojochoshiota address 1 of 2475 (72) inventor Shuzo Sakai Okayama Prefecture Akaiwa-gun Seto-cho, Ejiri Asahigaoka 1-chome No. 3 No. 41 (72) inventor Toshio Miyake Okayama Ishima-cho 1-chome No. 3 No. 23

Claims (10)

[Claims] The present invention relates to an aqueous solution containing sucrose and lactose, which 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 obtained by collecting a lactosucrose-rich liquid of at least 10%. 2. An aqueous solution containing sucrose and lactose is reacted with a glycosyltransferase, and the resulting lactosucrose-containing sugar solution is subjected to column chromatography, invertase using an alkali metal or alkaline earth metal type strongly acidic cation exchange resin. 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 lactosucrose-rich liquid obtained by collecting a 5 w / w% or more lactosucrose-rich liquid and collecting the same. 3. The lactosucrose-rich material according to claim 1, wherein the lactosucrose-rich material is a liquid or a solid. 4. An aqueous solution containing sucrose and lactose is reacted with a glycosyltransferase to remove contaminating saccharides from the obtained lactosucrose-containing saccharide solution to obtain 45 w / w saccharide composition.
% Of a lactosucrose-rich liquid, and collecting the lactosucrose-rich liquid. 5. An aqueous solution containing sucrose and lactose is reacted with a glycosyltransferase, and the resulting lactosucrose-containing sugar solution is subjected to column chromatography, invertase using an alkali metal or alkaline earth metal type strongly acidic cation exchange resin. 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 lactosucrose-rich product, which comprises collecting a 5 w / w% or more lactosucrose-rich liquid and collecting the liquid. 6. The method according to claim 4, wherein the lactosucrose-rich material is a liquid or a solid. 7. An oral or tube ingestion containing the high-lactosucrose content according to claim 1, 2 or 3. 8. The oral or tube feed according to claim 7, wherein the lactosucrose inhibits the generation of putrefactive substances produced by putrefactive bacteria in the intestine. 9. The oral or tube ingestible product according to claim 7, which contains a mineral together with lactosucrose. 10. The oral or tube ingestion according to claim 9, wherein the mineral is calcium, magnesium, phosphorus or a salt thereof.