JP3401203B2 - Foods containing galactosyl trehalose - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a food product containing galactosyltrehalose. Foods containing galactosyltrehalose are expected to have physiological effects such as anti-cariogenicity, bifidobacteria growth action, calcium absorption promoting action, immunostimulation, and carcinogenesis inhibition by eating this.

[0002]

2. Description of the Related Art Conventionally, oligosaccharides are known to exhibit physiological effects such as anti-cariogenic activity, bifidobacteria growth activity, calcium absorption promotion activity, immunostimulation, and carcinogenesis inhibition. Galactosyl trehalose, which is a type of galacto-oligosaccharide, is a non-reducing oligosaccharide composed of galactose and two molecules of glucose.
It is known to be produced by the action of galactosidase (Carbohydrate reseach.199 (1990) 227-234). However, this method has a drawback that the yield based on the raw material trehalose is as low as 5.1 mol%. A method for efficiently producing galactosyltrehalose is not known, and therefore foods and drinks containing galactosyltrehalose have not been known so far.

[0003]

SUMMARY OF THE INVENTION In view of the above-mentioned situation, the present inventors have made earnest studies with the object of providing a food containing galactosyltrehalose, and as a result, the action of β-galactosidase from lactose and trehalose. Have found that galactosyltrehalose can be easily produced in high yield, and the present invention has been completed based on such findings.

[0004]

[Means for Solving the Problems] The present invention relates to a food containing galactosyltrehalose, and a food or drink containing galactosyltrehalose obtained by adding β-galactosidase to a food or drink containing lactose and trehalose,
Furthermore, the present invention relates to foods and drinks to which galactosyltrehalose produced from lactose, trehalose and β-galactosidase is added. The galactosyltrehalose contained in the food of the present invention is obtained by adding β-galactosidase to lactose or a raw material containing lactose and trehalose or a raw material containing trehalose. In the production of galactosyltrehalose, lactose or a raw material containing lactose,
In addition to milk, commercially available lactose, skim milk, lactose-containing substances such as whey can be exemplified. Examples of trehalose or a raw material containing trehalose include commercially available trehalose, yeast containing trehalose, trehalose fermentation liquid, trehalose-forming enzyme reaction liquid, and the like.
The origin of β-galactosidase is not particularly limited, and examples thereof include β-galactosidase derived from Aspergillus, β-galactosidase derived from yeast, and β-galactosidase derived from lactic acid bacteria. Further, not only the purified enzyme but also a crude enzyme can be used and can also be used as an immobilized enzyme.

In the present invention, the concentrations of lactose or a substance containing lactose and trehalose or a substance containing trehalose are not particularly limited as long as they are concentrations at which galactosyltrehalose is substantially produced. As the lactose or the lactose-containing material, a solution prepared to have a lactose concentration of 1 to 50% by weight is used, and for the trehalose or the trehalose-containing material, a solution prepared to have a trehalose concentration of 1 to 50% by weight is used. The blending ratio of lactose and trehalose is 0.2 to 5 trehalose, preferably 0.5 to 2.
It is 0. The amount of β-galactosidase used is not particularly limited, but is 1 to 1000 units / ml, preferably 10 to 600 units / ml. The enzymatic reaction is p
H3-9, 10-80 degreeC, It is preferable to carry out at 20-50 degreeC for 0.5 hour to several days.

The above-mentioned enzymatic reaction causes a transfer reaction in which the galactose group of lactose is transferred to trehalose, and galactosyltrehalose is produced. The galactosyltrehalose produced by the transfer reaction of the present invention is O-β-
The main component is D-galactopyranosyl- (1,6) -O-α-D-glucopyranosyl- (1,1) -α-D-glucose. In addition, the galactose group is 4 of trehalose.
Galactosyltrehalose bound at position 3 or 3 is produced. The reaction solution containing the galactosyltrehalose thus produced can be used as it is, but it is also possible to obtain the galactosyltrehalose-containing solution after deactivating the enzyme by temperature treatment. Also,
This galactose group transfer reaction from lactose proceeds even in the presence of ethanol, and galactosyltrehalose can be produced even in an ethanol drink or the like. In the reaction in the presence of ethanol, the hydrolysis of lactose proceeds more slowly than in an aqueous solution, and galactosyltrehalose can be efficiently produced by the transfer reaction. In this case, the concentration of ethanol in the enzyme reaction system is 30% or less.

The galactosyltrehalose-containing liquid thus obtained by the transfer reaction can be directly used as food and drink, but galactosyltrehalose is separated from this liquid by a known separation means, or this liquid is As it is, it is added to other food ingredients and mixed, and biscuits containing galactosyltrehalose, candy, chewing gum, chocolate, snacks such as snacks, breads, noodles such as udon, soba, ramen, bifu, somen (improved noodles are also available. ), Canned bamboo shoots, asparagus, mushrooms, nameko and other vegetables, canned mandarin oranges, apples, figs, pears, canned fruits, garden products such as marmalade, jam, dried fruits and dried vegetables, tofu and its processing Products, soy protein, agricultural products such as margarine, alcoholic beverages, miso, natto, pickles, soy Fermented foods such as, condensed milk, milk powder, dairy products such as butter, cheese, ice cream, lactic acid bacteria products such as yogurt, ham, sausage, livestock foods such as canned meat, kamaboko, chikuwa, fried kamaboko, rice cake, fish sausage, etc. It is also possible to produce seafood products such as seafood products such as fish paste products, sardines, grated herring, shark fins, kazunoko, tatami sardines, sardines, dried shrimps and dried abalone. Hereinafter, the present invention will be specifically described with reference to Reference Examples and Examples for producing galactosyltrehalose, but the present invention is not limited to these examples.

[0008]

[Reference Example 1] 50 mM phosphate buffer pH 7.0, lactose 2.4 g, trehalose 2.5 g, Escherichia coli-derived β-galactosidase 4000 units, and reaction solution 10 ml were used.
The reaction was carried out at a temperature of 35 ° C. for 14 hours. H after the reaction
It was analyzed by PLC. The HPLC pattern is shown in FIG. The concentration of galactosyltrehalose produced as the main component was about 80 mg / ml (21.8 mol% based on the raw material trehalose).

[0009]

[Reference Example 2] The reaction-completed solution containing galactosyltrehalose produced in Reference Example 1 was passed through an activated carbon column,
Disaccharides or more sugars were adsorbed, washed with purified water, eluted with 50% ethanol, concentrated, and fractionated by HPLC. For preparative separation, use a polyamine column (YMC pack ^™
Polyamine 10mm × 250mm, YMC
Manufactured by K.K.), eluent acetonitrile: water = 65: 35, and flow rate 5 ml / min. A peak having a retention time of 13.5 minutes to 15 minutes was collected, and this fraction was concentrated to obtain 710 mg of a white powder (19.3 mol% based on trehalose as a raw material). ¹ H N of white powder obtained as described above
According to various spectra of MR and ¹³ C NMR, the carbon at the 1-position of the galactose group (shift value 106.15 ppm)
Is the 6th carbon of trehalose (shift value 70.97p
It was revealed from the coupling constant of hydrogen at the 1-position of galactose (J = 3.5 Hz) that it was β-bonded to pm).

When the white powder was hydrolyzed with 0.05N sulfuric acid, 2 mol of glucose was obtained based on 1 mol of galactose. Furthermore, galactose, glucose and trehalose were detected in the partial hydrolysis. In addition, as a result of examining the reducing property of this white powder by the Somogene Nelson method, the reducing property was not shown. From these results, this white powder is O-β-D-
It was galactopyranosyl- (1,6) -O-α-D-glucopyranosyl- (1,1) -α-D-glucose. The ¹³ C NMR spectrum of this white powder is shown in FIG.

[0011]

[Reference Example 3] 40 μl of 1M MES buffer pH 6.0
l, 0.5M lactose and trehaltose solution 250μ
1, 1.9 unit / ml β-galactosidase solution derived from E. coli, 10 μl of ethanol, 25 μl of ethanol, and 75 μl of purified water were used, and the reaction was carried out at a temperature of 30 ° C. for 23 hours. The sugar solution after the reaction was analyzed by HPLC. As a result, the amount of galacrosyl trehalose produced was 34 mM.

[0012]

Example 1 Lactose (48 g) and trehalose (50 g) were added to 200 ml of water to dissolve it, and the pH was adjusted to 7.0.
Add 8,000 units of β-galactosidase and add 35
It was performed at 24 ° C. for 24 hours. After the reaction, the enzyme was deactivated, and 20 g of a galactosyltrehalose fraction was obtained by activated carbon column chromatography and preparative HPLC. 98 g of galactosyltrehalose prepared above was added to 1000 ml of fresh milk containing 15 g of skim milk powder prepared separately.
Sterilized at ℃ for 3 minutes. This prepared milk was inoculated with 4 ml of a starter consisting of Lactobacillus bulgaricus 10 ⁸ / ml and Streptococcus thermophilus 10 ⁹ / ml, and cultured at 40 ° C for 9 hours. The yogurt obtained has a smooth texture, mild flavor and pH
It was possible to produce a galactosyltrehalose-containing yogurt having 4.7, and it was a preferable food product as yogurt.

[0013]

Example 2 53 parts by weight of wheat flour, 3 parts by weight of galactosyltrehalose were mixed with 8 parts by weight of shortening and 26 parts by weight of sugar, and 1 part by weight of egg, 1 part by weight of baking powder and 8 parts by weight of water were added and kneaded well, It was molded and baked at 180 ° C. for 10 to 15 minutes to obtain a galactocyltrehalose-containing biscuit.

[0014]

Example 3 20 parts by weight of unsalted butter is dissolved in a cream, 18 parts by weight of sugar is added and mixed, 19 parts by weight of egg is further mixed, and a small amount of vanilla essence is added and well mixed. To this, 40 parts by weight of soft flour, 1 part by weight of baking powder, and 2 parts by weight of galactosyltrehalose were added and kneaded well. This dough was molded and baked in an oven preheated to 170 to 180 ° C for 10 to 15 minutes to obtain a galactosyltrehalose-containing cookie.

[0015]

Example 4 Vinyl acetate resin (average degree of polymerization 650) 20
Parts by weight, polyisobutylene (average molecular weight 300,00
0) 30 parts by weight, ester gum 5 parts by weight, wax 20
Parts by weight were mixed at 110 to 120 ° C., and after the gum base was melted, mixed for about 1 hour. Then, 7 parts by weight of fatty acid monoglyceride, 6 parts by weight of acetylated monoglyceride and 12 parts by weight of calcium carbonate were added and mixed for 15 minutes, and then cooled to room temperature to obtain a gum base. This gum base (25 parts by weight), powdered sugar ( 51 parts by weight), glucose (5 parts by weight), starch syrup (12 parts by weight), glycerin (1 part by weight), fragrance (1 part by weight), galactosyltrehalose (5 parts by weight) were mixed and stirred for 15 minutes, and then rolled and cut into galactosyl. A chewing gum with trehalose was obtained.

[0016]

Example 5 14.5 parts by weight of milk, 1 part by weight of galactosyltrehalose, 0.1 part by weight of yeast food, 1 part of salt
Mix well, 6 parts by weight of sugar, 6 parts by weight of egg. This mixture is uniformly mixed with 58 parts by weight of well-sieved wheat flour (strong flour). Add 12 parts by weight of water to 1 part of dry yeast
A suspension of 1 part by weight and 0.4 part by weight of sugar is added and mixed. After the powder is gone, 20 parts by weight of butter are added in small amounts so that the whole blends in and mixed, and the mixture is mixed evenly and then kneaded for several hundred times. The dough thus obtained is fermented at a temperature of about 30 ° C. for about 1 hour 30 minutes. Divide this dough and roll it so that the surface is smooth. This divided and rolled dough is aged at 30 ° C. for 15 minutes. Roll the dough into thin triangles with a rolling pin, roll from the widest side, and place it on a thinly oiled plate with the seam down,
Roast for 0 minutes. Apply egg yolk and bake at 180 ° C for 15 minutes in a preheated oven. Thus, butter roll containing galactosyl trehatose was obtained.

[0017]

INDUSTRIAL APPLICABILITY According to the present invention, galactosyltrehalose in which the galactose group of lactose is transferred to trehalose can be produced in high yield. The resulting galactosyltrehalose is expected to have a bifidobacteria-proliferating effect of conventional galactosyloligosaccharides, an effect of reducing the content of cholesterol and neutral fat, and an effect of suppressing tooth decay, and various foods containing galactosyltrehalose can be produced. . In addition, trehalose is expected to have enzymes such as storage stability of proteins and proteins, and drying and freezing of foods and drinks.

[Brief description of drawings]

FIG. 1 is an HPLC profile of a sugar solution after the reaction in Reference Example 1. The peak at 10.4 minutes shows glucose, the peak at 15.9 minutes shows lactose and trehalose, and the peak at 29.3 minutes shows galactosyltrehalose.

FIG. 2 shows galactosyltrehalose in Reference Example 2.
^{It is a 13} C NMR spectrum.

Front Page Continuation (72) Inventor Yuko Konai 1480-2 A-404 Nogayacho, Machida City, Tokyo (56) References Carbohydrate Research Arch, Vol. 199 (1990), p.227-234 (58) Fields investigated (Int.Cl. ⁷ , DB name) A23L 1/30 A23G 3/00-3/30 A23L 1/09 C12P 19/14 CA (STN) JISC file (JOIS)

Claims (4)

(57) [Claims] 1. A raw material containing lactic acid or lactose and trehalo
Β-galactosider as a raw material containing sucrose or trehalose
The reaction mixture produced by the reaction of
Garakuro separated and purified by the Sonson method until its reducing property becomes negative.
A lactic acid bacterium product containing siltrehalose. 2. The lactic acid bacterium product is yogurt.
1 lactic acid bacteria product. 3. A raw material containing lactic acid or lactose and trehalo
Β-galactosider as a raw material containing sucrose or trehalose
The reaction mixture produced by the reaction of
Galacto separated and purified until the reducing property becomes negative by the Sonson method
Chewing gum containing siltrehalose. 4. A raw material containing lactic acid or lactose and trehalo
Β-galactosider as a raw material containing sucrose or trehalose
The reaction mixture produced by the reaction of
Garakuro separated and purified by the Sonson method until its reducing property becomes negative.
Biscuits, cookies containing siltrehalose and
A food selected from the group consisting of breads.