JP2001258489A - Method for producing wheat flour containing low allergen - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing wheat flour containing a low allergen by decomposition-treating the allergen in the wheat flour by using enzymes. SOLUTION: This method for producing the wheat flour containing the low allergen is characterized by mixing the wheat flour with a carbohydrase for hydrolysis-treating carbohydrate-based allergen contained in the above wheat flour and then treating with a protease for decomposing protein-based allergen.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for producing allergen-reduced wheat flour, and more particularly to a method for producing enzyme-treated allergen-reduced wheat flour.

[0002]

2. Description of the Related Art In recent years, particularly in developing countries, the number of patients with allergies tends to increase. Allergy is defined as a state of hypersensitivity in which a physiologically exaggerated, sometimes fatal, immune response occurs. Food allergies are sensitivities caused by consuming food, more precisely by ingesting certain components called allergens. In many cases,
These allergens are proteins. It is well known that continuous ingestion of allergens in eggs, milk, soy and wheat often causes allergies.

[0003] Wheat allergy is an allergic reaction that occurs in those who continue to consume wheat foods, including bread, noodles, cookies, cakes and even alcoholic beverages. The main symptom commonly observed is atopic dermatitis, which occurs shortly after ingestion of wheat products, usually resulting in breakouts and itching if the patient does not stop eating. Therefore, wheat allergy is a serious problem in many countries that have long had a habit of consuming wheat.

Although phylogenetic data showing a rapid increase in wheat allergy are not officially presented, such situations are often discussed in the field of allergology and the number of wheat allergies is increasing. There are many opinions.

[0005] Against this background, the major allergen in wheat is low molecular weight glutenin in gluten, which is identified as Gln-Gln-Gln-Pro-Pro-Pro as an IgE-binding antigenic determinant. It is known to have a motif.

[0006]

However, it has been difficult to decompose the sugar chain portion because allergens have several sugar chain proteins belonging to the α-amylase inhibitor family.

[0007] Therefore, an object of the present invention is to provide a method for producing an allergen-reduced wheat flour by decomposing an allergen in the flour by using an enzyme.

[0008]

Means for Solving the Problems In order to achieve the above object, the present inventors have found an enzyme capable of efficiently decomposing gluten in an allergen, and have applied this enzyme treatment to flour under appropriate conditions. As a result, they have found a method for producing the allergen-reduced flour of the present invention. That is, a two-step method using carbohydrase and protease is proposed to produce a low-allergenized wheat food that can be taken by most wheat allergic patients.

[0009] In the method for producing a low-allergen wheat flour according to the present invention, the carbohydrate allergen contained in the wheat flour is mixed with flour and carbohydrase, and then the protease is mixed to degrade the protein allergen. It is characterized by doing.

[0010] In a preferred embodiment of the present invention, the carbohydrase comprises at least one selected from the group consisting of cellulase and mannanase.

In a preferred embodiment of the present invention, the protease comprises at least one selected from the group consisting of actinase and alkaline protease.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION It is preferable that the enzymes used in the present invention have a low starch hydrolyzability in order to prevent the use of these enzymes from losing the inherent properties of flour. Therefore, the starch hydrolysis activity of the enzyme is also a guide when selecting the enzyme.

Test for Starch Hydrolysis Activity Potato starch (1 g) was gelatinized in water (100 ml) at 100 ° C. for 20 minutes and then cooled to 37 ° C. Starch solution (1m
L) was hydrolyzed at 37 ° C. for 30 minutes using the following enzymes (1 mg). The produced reducing sugar was quantified using 3,5-dinitrosalicylic acid by the method of Luuchsinger and Cornesky (1962). Starch hydrolysis activity was expressed as glucose equivalent μmol / min / mg enzyme.

As the enzyme, actinase AS (250 Tyr u
nits / mg solid, Kaken Pharma), Bromelain (1030 uni
ts / mg solid, Wako Pure Chemical Industries, collagenase (Clostridium hi
stilyticum-derived, 150 units / mg solid, Wako Pure Chemical Industries, transglutaminase (Streptoverticillum sp., derived, 3 units / mg solid, Ajinomoto), cellulase (Trichoder
ama viride, derived, 86.8 units / mg solid, Amano Pharmaceutical),
Seobiase (105 units / mg solid, Amano Pharmaceutical) and hemicellulase (98.6 units / mg solid, Amano Pharmaceutical) were used. The results are shown in Table 1.

[0015]

[Table 1]

As a result of Table 1, it was determined that the enzymes used in the present invention preferably include actinase, bromelain, collagenase, transglutaminase, cellulase, seobiase and hemicellulase.

Enzyme Treatment In the method for producing low-allergen wheat flour according to the present invention, first, carbohydrate allergen contained in wheat flour is hydrolyzed with carbohydrase. This is to avoid carbohydrase inactivation by proteases.

The carbohydrase is not particularly limited as long as it can hydrolyze a carbohydrate allergen contained in flour, and examples thereof include cellulase, mannanase, and hemicellulase. From the viewpoint of reducing allergens, preferred are cellulase and mannanase.

The reaction temperature between flour and carbohydrase is 45-55 ° C, more preferably around 50 ° C. The reaction time is 0.5 to 3 hours, preferably 1 to 3 hours.
2 hours. The ratio of carbohydrase to flour is enzyme: flour = 0.2 to 1: 1, preferably enzyme: flour = 0.4 to 0.6: 1.

Next, a proteolytic treatment is performed to further degrade the proteinaceous allergen. That is, in order to further decompose the wheat flour hydrolyzed by carbohydrase, the protease is subjected to proteolytic treatment.

The protease is not particularly limited as long as it can decompose the allergen. Preferably, actinase, alkaline protease and the like can be mentioned. These have high allergen-reducing ability, low cost, low cross-reactivity with serum, and low starch hydrolytic ability (Table
From 1).

After the carbohydrase treatment, the obtained reaction mixture and protease are mixed and reacted. The protease reaction temperature is preferably lower than the carbohydrase reaction temperature. The reaction temperature with the protease is, for example, 35 to 45 ° C, and preferably around 40 ° C.

The reaction time is 0.5 to 3 hours, preferably 1 to 2 hours. The ratio of protease to flour is enzyme: flour = 0.2-1: 1, preferably enzyme: flour = 0.4-0.6: 1.

Confirmation of Allergenicity of Wheat Flour An immunoassay was performed to check the allergenicity of flour obtained by various enzyme treatments.

The enzymatically treated samples (25 mg each) were
M urea (UTH buffer) containing 0.1 M Tris-HCl buffer (pH 8.6,
1 mL). The extract (0.1 ml) was evaluated by ELISA. Biotinylated anti-human IgE (Kirkegaad & Perry Lah. I
nc), streptavidin-peroxidase complex (Ro
che Diagnostics Co.) and o-phenylenediamine (Wa
ko Pure Chemical Ind.) was used for the measurement. Blood of eight wheat allergy outpatients was used for the measurement. Raw flour (control, 2.5 mg) and enzyme (2.5 mg) were tested in the same manner. Allergen, relative to control 1
Expressed as SA value. Table 2 shows the results.

[0026]

[Table 2]

As a result, actinase, collagenase,
Transglutaminase, cellulase, seobiase,
Hemicellulase has been found to be preferred as the enzyme.

Components of Low-Allergen Flour In order to examine the components of low-allergen flour (hereinafter also referred to as low-allergen batter), enzyme-treated low-allergen wheat flour was subjected to gel filtration.

[0029] Hypoallergenic batter (1 g) is added to water or UT
The H buffer (100 ml each) was extracted and the protein content of the extract was measured using a protein assay kit (Bio? Rad Lab). Solubilized proteins were expressed as the ratio of water soluble protein to UTH buffer soluble protein.

Gel filtration was performed as follows. Cellulase-water extract from flour treated with actinase (10 μL)
After concentration, a polyhydroxymethacrylate column (Jap
AnSpectroscopic Co., Biofine PO-4, 7.5 × 300 mm) and eluted with water at a flow rate of 1 mL / min. Eluate at 220 nm
The absorbance was measured. Vacitracin A (MW 1422), fluorescein isothiocyanate-glutaminyl glutaminyl glutaminyl prolyl proline (MW967), glutathione (MW 307) and lusyl glycine (MW188) were used as molecular weight markers. The results are shown in Figure 1. As a result, it was found that the reduced-allergen wheat flour contains at least starch, oligopeptide, and oligosaccharide.

Processing of allergen-reduced wheat flour into food The allergen-reduced wheat flour obtained according to the present invention cannot be used in principle like ordinary wheat flour. The allergen-reduced flour is batter-like, and gelatinization of starch in flour or addition of a surfactant has an advantageous effect on food processing. By reducing the allergen of flour,
The dough forming ability is lost.

Gelatinization of low-allergen wheat flour mainly includes partial gelatinization and complete gelatinization. In the case of partial gelatinization, for example, in a water bath at 50 ° C to 75 ° C for 15 minutes to
There is a method of heating for 45 minutes. For complete gelatinization, for example, there is a method of steaming for 15 to 30 minutes.

The allergen-reduced flour thus obtained can be processed into foods such as cupcakes, noodles, pizzas, cookies and wafers.

[0034]

Here, one embodiment of the present invention will be described. However, the present invention is not construed as being limited to the following embodiment.

Example 1 Commercial flour (Showa Sangyo, trade name: Cleopatra) was used as a starting material. Allergenicity of flour when cellulase and actinase were used as enzymes was examined.

In the one-step process, flour (100 g) was mixed with an aqueous cellulase or actinase solution (60 ml) and the mixture was incubated at 50 ° C. for cellulase and 40 ° C. for actinase.

In a two-stage process, the volume of water in the first stage
(55 ml), and the same operation was performed. The remaining water (5 ml)
It was used to dissolve the enzyme used in the second step. Table 3 shows the results
Shown in

[0038]

[Table 3]

As a result, when cellulase was used as the first step (hydrolysis of carbohydrate allergen) and actinase was used as the second step (decomposition of allergen), the reaction time was 1 hour and the enzyme / flour ratio was , 0.5% was sufficient.

Example 2 The allergen-reduced wheat flour that had been subjected to the enzyme treatment was partially gelatinized or completely gelatinized and used for food processing.

The allergen-reduced batter (100 g each) was partially gelatinized by heating in a water bath at 65 ° C. for 30 minutes. Further, the mixture was completely gelatinized by steaming at 65 ° C. for 20 minutes in a water bath. The heat-treated batter sample was subjected to viscosity measurement, gelatinization degree and physical property measurement.

Regarding the viscosity, a low allergenic flour batter (30 g) was mixed with a rapid viscometer (Newport Scient
ific) cup and measure the change in viscosity of the sample.
It was measured by the method of nbaugh et al. (1989). The sample temperature was maintained at 65 ° C. The result is shown in figure 2.

Β-amylase pullulanase method (Kainuma
Et al., 1981). Partially and completely gelatinized batter samples (30 mg each) were suspended in water (8 ml). The suspension was divided into two portions, one of which was completely gelatinized with 1NNaOH and then hydrolyzed with an enzyme, and the reducing sugar during hydrolysis was measured by the method of Luuchsinger and Cornesky (1962). . The gelatinization degree was expressed as a ratio of values before and after alkali gelatinization.

The allergen-reduced flour batter was placed in an alumina cup (diameter 38 mm, height 13 mm) and steamed for 10 minutes to gel. The gel was cut to a height of 13 mm. Gel hardness, cohesiveness and adhesiveness
(13mm diameter) texture measuring instrument (General Food
s, GTX-2). Clearance, sample temperature and bite speed, 2mm, 65 ° C and 6 times, respectively
/ Min. Pizza was measured under similar conditions. The cupcake was cut into a cube (20 × 20 × 20 mm). For each cube, the clearance was set to 4 mm, and the hardness and adhesion were measured at 25 ° C. using a nickel plunger (50 mm in diameter) in the same manner.

Example 3 Flour (100 g) was mixed with an aqueous cellulase solution (55 ml),
The mixture was incubated at 50 ° C. Thereafter, an actinase aqueous solution (5 ml) was added, and the reaction was performed at 40 ° C. for 1 hour.
A low allergen flour was prepared. Various foods were manufactured using the allergen-reduced flour. Table 4 shows the ingredients of the food.

[0046]

[Table 4]

Low allergen margarine derived from palm oil,
Tapioca starch, stearic acid sugar ester (SE S-167
0) was provided by Asahi Denka Kogyo, Showa Sangyo and Kao.
The reagents used were of reagent grade. Other materials were purchased from the market.

First, a method of processing each food will be described. In noodles, batter completely gelatinized, 65-70 ℃
And the mixture was extruded on a spaghetti maker (Yasukawa Electric Mgf.). For the cupcakes, the material mixture (20 g each) was placed in an alumina cup (6 cm diameter) and steamed for 7 minutes. For the pizza, the ingredient mixture (130 g each) was cast on a plate (18 cm diameter) and baked at 170 ° C. for 10 minutes. For cookies, the material mixture was molded into a diameter of 3 cm and a thickness of 5 mm and baked at 160 ° C. for 7 minutes. In wafer, water (50mL)
, Butter (10 g) was suspended therein, and first, a part of the allergenized flour was heated to another 90 ° C, and other ingredients were blended. Test oven with two plates (156 x 110 mm each) (Heb
The wafer was fired at 130 ° C. using enstrit Co, model 965) to obtain a wafer. For puffed food, use noodles cut into grits, and noodle grits (10 g each) are molded (diameter).
Rice cake making machine (We-Pop Co, mode 10cm, depth 1cm)
l AP-01).

A texture measuring instrument (compression speed 1 mm / sec, polyacetal plunger (diameter 5 mm)) (Yamaden Co.,
Rheoner RE 3305) was used to measure the breaking strength of cookies, wafers and puffed foods.

The specific volume was measured by the rapeseed replacement method (A
American Association of Cereal Chemists, 1983). The specific volume was expressed as volume (mL) / weight (g). Table 5 shows the results.

[0051]

[Table 5]

As a result, it was found that using the partially gelatinized butter, the cake had a higher specific volume of 3.8 (Table 5). The addition of surfactant was preferred for cupcakes, pizzas and wafers.

[0053]

The allergen-reduced wheat flour of the present invention has an advantageous effect that the allergenicity can be extremely reduced by a simple enzyme treatment without greatly changing the characteristics of the raw wheat flour.

Further, the allergen-reduced wheat flour of the present invention comprises:
Since the dough characteristics of wheat flour are supplemented by performing an appropriate gelatinization treatment, there is an advantageous effect that it can be easily used for food processing.

[Brief description of the drawings]

FIG. 1 shows a pattern of gel filtration of a water-soluble fraction of flour treated with cellulase and actinase.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (reference) A21D 13/00 A21D 13/00 13/08 13/08 A23L 1/16 A23L 1/16 A F-term (reference) 4B018 LB01 LB02 MD22 ME07 MF12 4B023 LC09 LE26 LK17 LP16 4B032 DB05 DB21 DB28 DB32 DG02 DK51 DL20 DP01 4B035 LC06 LC09 LE01 LG35 LG51 LP41 4B046 LA06 LC06 LG46 LG46

Claims (3)

[Claims] 1. A low-allergenized wheat flour, comprising mixing wheat flour and carbohydrase to hydrolyze a carbohydrate allergen contained in the wheat flour, and then mixing a protease to degrade a protein allergen. Production method. 2. The method according to claim 1, wherein the carbohydrase comprises at least one selected from the group consisting of cellulase and mannanase. 3. The protease according to claim 1, wherein the protease is at least one selected from the group consisting of actinase and alkaline protease.
3. The method according to claim 1, wherein the method comprises a seed.