WO2000024267A1 - Gelatins with low allergenicity - Google Patents

Abstract

Gelatins for edible, medicinal or cosmetic use originating in fowl skin, bone and/or tendon, characterized by having an isoelectric point of 7 to 10 and undergoing no or little antigen-antibody reaction with the serum of a patient with gelatin allergy; and peptides obtained by hydrolyzing the gelatins. Because of having low allergenicity, these gelatins and peptides can be safely supplied to patients with gelatin allergy. Thus, they are useful as gelatins to be employed in the fields of foods, drugs or cosmetics with a strong need for safety.

Description

 Description Low allergenic gelatin Technical field

 The present invention relates to hypoallergenic gelatins and peptides. More specifically, the present invention relates to gelatin and peptides which are used for food, medical use (including pharmaceutical use) and cosmetics and do not cause allergic symptoms. Conventional technology

 Gelatin is a protein that is most contained in vertebrates, collagen, which is denatured by physicochemical treatment to make it soluble, and then purified and dried. Thus, gelatin is produced from collagen-containing tissue. Examples of collagen-containing m¾ include the skin, bone, cartilage, tendons, placenta, etc. of animals (eg, cows, pigs, rabbits, sheep, chickens, etc.). Inexpensive cow and pig bones and hides are used.

 Conventionally, gelatin has been produced by a method in which raw materials are pretreated with alcohol or acid and then extracted with warm water. In recent years, pretreatment with an enzyme (protease) has been studied from the viewpoints of efficiency and high purity.

 The alkali pretreatment is a method in which the raw material is immersed in a 1-5% lime solution for 30-100 days and then washed with neutralized water. This removes the impurities and makes the gelatin readily extractable with warm water. Since the amide group of the side chain of arginine, glutamine, and asparagine is carboxylated during alkali treatment, the isoelectric point of alkali-treated gelatin is 4 to 5.5, which is lower than that of collagen.

Pretreatment with acid is a method in which the raw material is immersed in a dilute acid solution such as sulfuric acid or hydrochloric acid for about 1 to 2 days, and then washed with water. The acid-treated gelatin has an isoelectric point of 7 to 10, usually 7.5 to 9.5, which is almost the same as collagen. Isoelectric point of enzyme-treated gelatin is also acid-treated zera It is almost the same as Chin.

 Gelatin has many properties such as foaming properties, film-forming ability, water retention, protective colloid properties, elasticity, thermoreversible sol-gel conversion properties (gelling properties), and is available inexpensively and in large quantities. Therefore, it is used in a wide range of applications such as food, medical, cosmetics, photography, and industrial use.

 As an edible product, it is used as a raw material for jelly that makes use of its gelling properties. In addition to gelatin, vegetable polysaccharides such as agar, carrageenan, and pectin have gelling properties, but all are vegetable, have poor digestion and absorption, and have low nutritional value. On the other hand, gelatin is a typical animal protein with good digestion and absorption, and has an excellent texture when using jelly.

 Furthermore, it is used as a raw material for marshmallows by utilizing its foaming properties. In addition, it is widely used as a primary or secondary raw material for gummy candy, yogurt, ham, sausage, soup, nokubaroa, ice cream, etc.

 For medical use, it is applied to hard and soft capsules. These depend on the various properties of gelatin, such as viscosity, gelling properties, and film-forming ability.

 It is also used as a poultice because of its excellent affinity for skin, water retention and adhesiveness. Other applications include binders for tablets and iLiiL materials.

 For cosmetics, it is incorporated into emulsions and packs as moisturizing ingredients.

 For photographic purposes, it is used as a binder for silver halide as a photosensitive material. A substance that can replace gelatin in terms of its gelling properties and protective colloid properties has not yet been developed. For industrial use, it is used as an adhesive for woodworking such as plywood and furniture.

Peptides obtained by hydrolyzing gelatin with acid, alkali, oxygen or heat are also widely used for food, medical use and cosmetics. For food use, it is more soluble than gelatin and has better absorption and absorption, so it is added as a source of amino acids such as nutrients. Utilizing its protective colloid properties, it is also used as an agent for lowering alcoholic beverages such as sake. For medical use, it is used in plasma substitutes, vaccine stabilizers, etc. For cosmetics, it is used in hair care ingredients. Gelatin is very similar in molecular structure irrespective of species, and it has been considered that even heterogeneous ones have little or no antigenicity. Edible gelatin has been evaluated as `` Class A (1), ADI not specified '' by the FA OZWH O Joint Food Additives Expert Committee, and has been listed as a pharmaceutical product by the Japanese Pharmacopoeia, three departments. The safety of humans in humans is widely accepted.

 However, in recent years, the number of infants who cause an allergic reaction with gelatin-containing ヮ Kutin ゃ foods is increasing rapidly. It has been reported that about 1 in 500 vaccine recipients causes immediate-type and 1 in 110 cases causes non-immediate-type gelatin allergy (Ministry of Health, Labor and Welfare, Japan) Vaccination Research Center: Comprehensive study on effective implementation and side effects of vaccination, 151-154, 1998). It has been pointed out that the main cause of this sensitization is a triple vaccine or a baby food containing gelatin. Gelatin is used for a wide variety of purposes, including food, medical, cosmetic, photographic, and industrial uses.However, when a patient who receives or consumes a gelatin-containing product is exposed to allergic symptoms. Can cause.

 Particularly severe gelatin allergies result from live vaccine vaccination. Raw vaccines required the addition of protein stabilizers, such as gelatin or its hydrolysates or human serum albumin, to prevent loss of titer. Human serum albumin is problematic for its widespread use, including the presence of unknown pathogens. On the other hand, saccharides such as lactose, sorbitol, glucose, purified sucrose and dextran did not provide a sufficient stabilizing effect. In recent years, the danger of gelatin-containing pectin has been widely recognized, and although gelatin-free vaccines have been developed, measles vaccines, which require a low titer and have a stable effect, are supplemented with evening protein stabilizers Is essential. Therefore, there has been a strong demand for the development of an alternative stabilizer or gelatin that functions as a stabilizer without causing allergic symptoms.

Although allergy to gelatin by vaccination has been recognized, little attention has been paid to gelatin allergy by using food, other pharmaceuticals, and cosmetics. Therefore, serious accidents may occur due to the use of these gelatin-containing products. Is feared. Therefore, it is socially important to avoid this before it occurs.

 Gelatin-containing foods include many everyday foods such as jelly, nopa loa, gummy candy, yogurt, nose, sausage, soups, and ice cream. In addition, gelatin hydrolyzed peptides are also used in foods, and in particular, foods containing large amounts of gelatin hydrolyzed peptides, such as beverages, have recently become commercially available. The content of gelatin hydrolyzed peptides in these foods may be 5 g at a time. This is enough to cause anaphylactic shock. Furthermore, gelatin is a main raw material for soft capsules and hard capsules for pharmaceutical use. Ingestion of these foods and medicines by patients with gelatin allergies may cause allergic symptoms such as itching in the mouth and eyes, nasal discharge, fever, vomiting, headache, diarrhea, dermatitis, and asthma, and in some cases anaphylactic shock. It can happen. —On the other hand, as described above, gelatin is an essential component for maintaining the shape, physical properties, taste, and the like of these foods and pharmaceuticals, and no inexpensive substances have been developed.

 In cosmetics, gelatin is used as a moisturizing component in emulsions and lotions. Gelatin is also used in cataplasms. The use of these cosmetics / pharmaceuticals in gelatin allergy patients may be accompanied by skin irritation such as itchiness, erythema, papules, vesicles, and surface detachment. There was no inexpensive alternative material that had the same strength, water affinity, and adhesion to the skin.

 As mentioned above, the use of gelatin in foods, pharmaceuticals, cosmetics, etc. often depends on its various properties, and most of them are irreplaceable uses, so they have the properties of conventional gelatin. The development of safe and inexpensive substances that do not cause allergic problems was desired.

From this point, attention has been paid to the antigenicity of the raw material itself, and gelatin with low antigenicity, which is mainly composed of type II collagen, has been developed (Japanese Patent Application No. 9-1210, Ryo 5). reference). However, it is not easy to obtain large amounts of raw materials containing type II collagen such as cartilage, which is expensive compared to cow or pig skin and bone, and has a low yield of gelatin, which poses a problem for commercial production. Many. In addition to the above-mentioned inventions, various inventions utilizing hydrolysis to remove the antigenicity of gelatin have been proposed (Japanese Patent Publication No. 62-36495, Japanese Patent Application Laid-Open No. 7-8). Japanese Patent Application Laid-Open No. 22999, Japanese Patent Application Laid-Open No. Hei 8-193939, Japanese Patent Application Laid-Open No. 9-176196, etc.). When hydrolyzing is performed, the properties specific to gelatin (eg, gelling properties, film-forming ability, etc.) are lost, and the range of use is extremely limited. Further, as shown in the examples described below, gelatin extracted from the skin or bone of a commercially available cow or pig has allergenic properties. Further, gelatin prepared by alkali treatment has allergenicity regardless of the type of raw material, and it is highly probable that allergenicity remains in the gelatin hydrolyzed peptide prepared in the above publication. .

 As described above, the present invention cannot be said to be a safe and inexpensive gelatin which has various properties of conventional gelatin which is the object of the present invention and which does not cause allergic symptoms, and a method for producing the same.

 In order to solve the above-mentioned problems, the present inventors have found that they have the same processing suitability and taste characteristics as conventional gelatin, and do not cause antigen-antibody reaction with the serum of a gelatin allergy patient or have a low antigen-antibody reaction. Yes, intensive studies have been conducted to develop a method for preparing safe gelatin at low cost and in large quantities. As a result, it was found that gelatin prepared from poultry skin, bone and / or tendon, preferably chicken skin, bone and bone or foot or foot without alkali treatment can achieve the intended purpose. Was. The present invention has been made based on such knowledge, and when used for food, medical use, cosmetics, etc., gelatin, which does not cause allergic symptoms to gelatin allergic patients, can be produced inexpensively and in large quantities. It can be provided. Disclosure of the invention

 The present invention is based on poultry skin, bone and Z or tendon, having an isoelectric point of 7 to 10, and not producing an antigen-antibody reaction with a serum of a patient with gelatine allergies or having a low antigen-antibody reaction. An edible, medical or cosmetic gelatin.

Further, the present invention relates to a gelatin allergy patient containing the above gelatin. It is a hypoallergenic food, medical or cosmetic product.

 Further, the present invention comprises immersing poultry skin, bone and bone or tendon in an acid solution, collecting the precipitate, heating and extracting the obtained precipitate with an acid solution, and purifying the extract. The above is a method for producing gelatin.

 Further, the present invention is an edible, medical or cosmetic peptide obtained by hydrolyzing the above gelatin and collagen or its precursor. BRIEF DESCRIPTION OF THE FIGURES

 FIG. 1 shows the gelatin of the present invention (chicken foot gelatin and chicken skin gelatin), the gelatin hydrolyzing peptide (chicken foot gelatin hydrolyzate), the gelatin for sale food, the gelatin for pharmaceuticals, the gelatin for cosmetics and the gelatin hydrolyzate ( FIG. 2 is a graph showing antigen-specific IgE antibody levels of serum of a patient diagnosed as having gelatin allergy (black) and serum of a healthy individual (white) with respect to (a gelatin hydrolyzate for medical use).

 Figure 2 shows antigen-specific IgE antibody levels in serum of patients diagnosed with gelatin allergy to acid-treated gelatin (black) or alkali-treated gelatin (white) prepared from chicken feet, cow skin or pig skin. It is. BEST MODE FOR CARRYING OUT THE INVENTION

 As described above, the gelatin of the present invention is gelatin derived from poultry skin, bone and / or tendon, has an isoelectric point of 7 to 10, and recognizes conventional gelatin as an allergen. It is characterized in that no antigen-antibody reaction occurs or the antigen-antibody reaction force is low when an antigen-antibody reaction is performed with serum. A low antigen-antibody reaction as described above means that when a statistically significant difference test is performed based on the test results, no significant difference is observed as compared to the serum of a healthy subject, and the condition is considered to be low. .

Poultry skin, bone and Z tendon are used as starting materials for the hypoallergenic gelatin according to the present invention. These raw materials can utilize by-products of the meat industry. Poultry can be exemplified by chickens, ducks, geese, turkeys, etc. It is profitable. Therefore, chicken skin, bone and Z or foot are preferably used as starting materials of the present invention. The origin of the gelatin of the present invention can be confirmed by an antigen-antibody reaction.

 The gelatin of the present invention is prepared using the above-mentioned raw materials. An important point in production is that no alkali treatment is performed. Many commercially available gelatins are pretreated with alkali, and as shown in the test examples below, allergenicity of gelatin is increased regardless of the animal species and location when treated with alcohol. Therefore, it is necessary to pay close attention to pretreatment.

 The presence or absence of the alkaline treatment can be confirmed by examining the isoelectric point of gelatin. The isoelectric point of acid-treated gelatin is about 7 to 10 which is almost the same as collagen, usually 7.5 to 9.5, whereas the isoelectric point of alkali-treated gelatin is that of arginine, glutamine and asparagine side chains. It is 4 to 5.5, which is lower than that of collagen because the amide group is carboxylated.

 In addition to the isoelectric point, the presence or absence of an alkaline treatment can be confirmed by detecting ornithine by amino acid analysis. When the alkali treatment is performed, a part of arginine is changed to ordinine. This can be confirmed by amino acid analysis because there are several residues in the 100,000 amino acid residues. On the other hand, ordinin is not detected in native collagen and acid-treated gelatin.

 More specifically, as an example of the method for producing the gelatin of the present invention, the above-mentioned raw material is immersed in an acid solution (a hydrochloric acid solution, a sulfuric acid solution, or the like, preferably about 3% hydrochloric acid solution) for about 2 days. Thereafter, the precipitate is collected and washed with water, and then an acid solution having a pH of about 4 (a hydrochloric acid solution, a sulfuric acid solution or the like, preferably a hydrochloric acid solution) is added, and the mixture is heated and extracted. The gelatin of the present invention can be obtained by filtering, desalting, and drying the obtained extract. Gelatin thus obtained may be further purified, if necessary, according to a conventional protein purification method.

As shown in the test examples described below, the gelatin of the present invention has a jelly strength equivalent to that of conventional gelatin. Furthermore, it has foaming properties, film-forming ability, water retention, protective colloid properties, elasticity, etc., and can be used for various applications in which conventional gelatin is used. It is particularly preferably used for food, medical use, and cosmetics.

 It is used for jelly, gummy candy, yogurt, ham, sausage, soup, bavarois, ice cream, and so on. By using the gelatin of the present invention, it is possible to produce a food with extremely low allergenicity without changing palatability.

 Capsules for medical use. It is used as a patch and vaccine stabilizer. The use of the gelatin of the present invention makes it possible to ensure the safety without changing the basic properties of conventional gelatin. It may also be used to detoxify toxins in vaccine production.

 For cosmetics, it is used in creams, ointments, and lotions. Since the gelatin of the present invention has no or very low allergenicity, it is possible to avoid skin irritation caused by gelatin contained in these cosmetics.

 As long as low allergenicity is ensured, the gelatin of the present invention may be mixed with other gelatin (eg, alkali-treated gelatin, gelatin derived from animal species other than poultry, etc.) and used. As a result, it is possible to maintain the processing suitability and the Z or taste characteristics, and it is possible to supply a more inexpensive product.

 The peptide of the present invention is a low-molecular substance obtained by hydrolyzing the above-mentioned gelatin according to a conventional method. Further, the peptide of the present invention does not produce an antigen-antibody reaction when it is subjected to an antigen-antibody reaction with a patient serum that recognizes conventional gelatin produced from bovine or pig bone and skin as an allergen. It is characterized by a low value.

 The peptide of the present invention can be obtained by hydrolyzing the gelatin of the present invention and collagen or a precursor thereof with an enzyme, an acid or the like according to a conventional method. The molecular weight can be appropriately selected depending on the application, but is preferably from 50,000 to 50,000.

The peptide of the present invention provides safety without changing the basic properties of the conventional gelatin hydrolyzate or collagen hydrolyzate. It can also be used for various applications. In particular, low molecular weight peptides are very well soluble in water and have no gelling properties even at low temperatures, such as foodstuffs such as lavenders, beverages, confectionery, stabilizers for vaccines, tablets, plasma substitutes, etc. It can also be used for pharmaceuticals such as and cosmetics such as shambhu. Industrial applicability

 The gelatin and the peptide of the present invention do not impair the characteristics of conventional gelatin and gelatin-degraded peptides, and have no or very low allergenicity. Therefore, it is useful for foods, pharmaceuticals, cosmetics, etc., where safety is particularly required in the fields of use of conventional gelatin and gelatin-decomposed peptides. Moreover, the skin, bone and Z or tendon of poultry, which is a raw material of the gelatin and peptide of the present invention, is a by-product of the meat industry, and since it can be used, resources can be effectively used. It can be manufactured at low cost. Example

 Hereinafter, the present invention will be described in more detail based on examples and test examples, but the present invention is not limited to these examples.

Example 1

Preparation of low allergenic gelatin (1)

 After the chicken legs were crushed, they were immersed in 5 times the volume of 3% hydrochloric acid solution. After 48 hours, the precipitate was collected and washed with water to remove excess acid. A 5-fold amount of hydrochloric acid solution (pH 4) was added, and the mixture was heated and extracted for 60 hours, applied to a mixed-bed ion exchange resin (Bio-Radonnay earth, AG501-X8) column washed with distilled water, and the pass fraction was collected. Then, freeze-drying was performed to obtain the hypoallergenic gelatin of the present invention. Example 2

Manufacture of gummy candy

Yogurt powder in water (14 parts per 100 parts by weight of the blended component mixture, and so on) (6 parts) was added and heated and dissolved, and sugar (25 parts), starch syrup (30 parts) and the low-allergenic gelatin (25 parts) obtained in Example 1 were added, and the mixture was boiled down for 20 minutes. This was filled into a suitable casing, molded, and then cooled to produce gummy candy. Example 3

Jelly production

 In hot water (85 parts), add the low allergenic gelatin (3 parts) obtained in Example 1 and sugar (10 parts) to dissolve, add sliced lemon juice (2 parts), and add for 20 minutes boiled. This was poured into a mold and cooled to produce jelly. Example 4

Manufacture of hard capsules

 After attaching a 5% solution of the low allergenic gelatin obtained in Example 1 to a forcepsell type pin, the solution was cooled and dried. When dried to a moisture content of 15 to 18%, the pin was pulled out, and further dried to a moisture content of 12 to 15% to produce a hard capsule. Example 5

Production of low allergenic gelatin (2)

 The hypoallergenic gelatin of the present invention was obtained in the same manner as in Example 1 except that chicken skin was used instead of the chicken feet of Example 1. Example 6

Preparation of low allergenic gelatin hydrolyzate peptide

10 g of the hypoallergenic gelatin obtained in Example 1 was dissolved in 500 ml of a 3% hydrochloric acid solution, followed by hydrolysis for 73 hours. After neutralization, this was applied to a mixed-bed ion-exchange resin (manufactured by Bio-Rad, AG501-X8) column washed with distilled water, and the pass fraction was collected and spray-dried to obtain the peptide of the present invention. . Example 7

Manufacture of juice drinks

 To water (56.8 parts) add citrate (0.2 part), orange juice (35 parts), sugar (5 parts) and gelatin hydrolyzed peptide obtained in Example 6 (3 parts). After heat-dissolving and cooling, the mixture was filled in a container and sterilized by heating at 85 ° C for 30 minutes to produce a fruit juice beverage. Example 8

Production of low allergenic gelatin (3)

 The hypoallergenic gelatin of the present invention was obtained in the same manner as in Example 1 except that duck skin was used in place of the chicken feet of Example 1. Test example 1

Proof of low allergenicity (1)

 The reactivity of the gelatin of the present invention prepared by the method described in Examples 1, 5 and 6, and the gelatin hydrolyzed peptide and the commercially available gelatin with the antigen-specific IgE antibody in the serum of a gelatin allergy patient was examined.

① Test sample

The above-mentioned gelatin and hydrolyzed gelatin of the present invention, and commercially available edible gelatin, pharmaceutical gelatin, cosmetic gelatin and pharmaceutical gelatin hydrolyzed peptide were used as test samples. Table 1 shows the properties of the test samples. The jelly strength was measured according to the method described in Japanese Industrial Standard JIS K 6503-1977. Test sample material Raw material strength (bloom)

 Chicken leg gelatin C Example 1) Chicken leg 4 1 4

 Chicken skin gelatin Example 5) Chicken skin 3 7 5

 ゼ ラ チ ン foot gelatin chicken foot without gelation

 Hydrolyzed peptides (Example 6)

 Commercially available edible gelatin beef skin 100 to 300

 Commercially available pharmaceutical gelatin bovine bone 2 8 4

 Gelatin for commercial cosmetics Beef bone, cowhide No gelation

 Commercially available pharmaceutical gelatin, beef bone, cowhide, not gelled

 Hydrolyzate

② Preparation of sample solution

 The gelatin, the gelatin hydrolyzed peptide and the commercially available gelatin of the present invention are diluted with PBS (phosphate-buffered physiological saline H7.2) so as to obtain a 0.1% (W / V) solution, and the resulting mixture is heated with hot water. The mixture was sufficiently stirred to dissolve. These solutions were used as sample solutions.

 ③ Serum of allergic patients

 A doctor collected a small amount of blood from 15 (6 males and 9 females) allergic patients who were diagnosed as gelatin allergy and showed RAST (radio allergosorbent test) positive. One used.

 血清 Serum of healthy person

 Umbilical cord blood was collected from 6 newborn babies (same number of men and women), and serum was separated and frozen and stored by a conventional method.

⑤Test method

 The reactivity (antigen-specific IgE antibody value) with the antigen-specific IE antibody in the serum of a gelatin allergy patient to each of the above sample solutions was examined by ELISA (enzyme-linke {i immunosorbent assay). Incidentally, ELISA was performed according to the method described in "Daimi Fujiwara et al., Immunology Research Method Book, 199-206, 1992, Chugai Medical Company, Tokyo".

 The operation outline of the ELISA method is shown below.

(1) Add the above sample solution to a 96-well ELISA microplate, and immobilize the antigen protein on the plate. (2) After washing the plate, block by adding human serum albumin to prevent nonspecific adsorption of the sample or labeled antibody.

 (3) After washing the plate, add allergic patient serum and healthy subject serum as samples to separate wells and react with antigen.

 (4) After washing the plate, add alkaline phosphatase-labeled anti-human IgE e-chain antibody and react.

 (5) After washing the plate, the substrate (Lumiphos 530; 4-Methoxy-4- (3-phosphate-phenyl) spiro [1,2-dioxeta e-3,2'-adamantane] disodium salt, And the amount of luminescence generated by the dephosphorylation reaction of alkaline phosphatase is measured. (6) Perform the measurement with a plate reader (LUMINOUS CT-9000D, Diayato K.K.). ELISA using serially diluted standard antibodies is performed simultaneously, and the antigen-specific IgE antibody concentration is calculated from the standard curve created by this.

⑥ Result

 The results of the above tests are shown in Figure 1 (all are mean soil standard errors). In addition, the solid black indicates the specific IgE antibody value of the serum of the patient with gelatin allergy for each sample solution, and the white outline indicates the specific IgE antibody value of the serum of the healthy individual.

As shown in FIG. 1, the antigen-antibody reaction against the gelatin and the gelatin-hydrolyzed peptide of the present invention and the commercially available gelatin was examined using the serum of a patient with gelatin allergy and the serum of a healthy subject. The specific IgE antibody reactivity of the serum of a gelatin allergy patient to the degraded peptide was low, and no significant difference was observed between the specific IgE antibody values of the serum of a healthy subject and those. On the other hand, in the case of commercially available edible gelatin, pharmaceutical gelatin, cosmetic gelatin, and pharmaceutical gelatin hydrolyzing peptide, the specific IgE antibody level of the serum of a patient with gelatin allergy was high for all of them, and the serum of healthy subjects against them was high. A significant difference was found between the specific IgE antibody value and the specific antibody. Test example 2 Proof of low allergenicity (2)

 To examine allergenicity due to differences in livestock species and production methods, prepare acid-treated gelatin or gelatin-treated gelatin from chicken feet, cowskin or pigskin according to the conventional method, and use the serum in patients with gelatin allergy to these gelatins. Was tested for its reactivity with antigen-specific IgE antibodies. The acid-treated gelatin derived from chicken feet refers to the gelatin of the present invention prepared by the method described in Example 1.

 ① Test sample

 Acid-treated gelatin or Arikari-treated gelatin prepared from chicken foot, cow skin or pig skin according to a conventional method was used as a test sample. Table 2 shows the number of orditin (in 100,000 residues) and the isoelectric point of the obtained gelatin.

 Table 2

 ② Preparation of sample solution

 It was prepared in the same manner as in Test Example 1.

 ③ Serum of allergic patients

 The same serum as in Test Example 1 was used.

 ④Test method

 The test was performed in the same manner as in Test Example 1.

 ⑤ Result

The results of the above test are shown in Figure 2 (average soil standard error). Black singular I g E antibody value of the gelatin allergic patients serum to acid-processed gelatin prepared from each raw material, white is the specific I _g E antibody value of Zerachinarerugi one patient serum against Al force re-processed gelatin prepared from the raw materials Show. As shown in FIG. 2, when the acid-treated gelatin of the animal species was examined, the specificity of the serum of a patient with gelatin allergy against the gelatin of the present invention was low. The Ig E antibody value was significantly higher.

 When alkaline treatment was performed as a pretreatment, the specific IgE antibody levels of allergic patient sera increased uniformly. Furthermore, in a test conducted separately, no significant difference was observed between the specific IgE antibody value of the serum of one patient with gelatin allergy against the gelatin of the present invention and the specific IgE antibody value of the serum of a healthy subject against these. Was.

 That is, almost no specific IgE antibody recognizing the gelatin of the present invention as an antigen (allergen) was found in the serum of a patient with gelatin allergy. However, gelatin prepared from chicken feet, which is a raw material of the gelatin of the present invention, by an alkali treatment showed a specific IgE antibody. Gelatin prepared from cow skin and pig skin had allergenic properties regardless of the pretreatment.

 The reason for the higher allergenicity of Al-treated gelatin compared to acid-treated gelatin is that the alkali treatment changes the primary structure of the gelatin molecule and loses sequence similarity to human collagen (gelatin). It is inferred.

 Although not shown, a sample solution was also prepared and tested for commercially available gummy candy, jelly, and hard capsules. The results were almost the same as those of commercially available gelatin. On the other hand, in Examples 2, 3, 4, and 7, gummy candies, jellies, hard capsules and fruit juice drinks made using the gelatin or peptide of the present invention were gelatin-allergic patients. Reactivity with antigen-specific IgE antibodies in serum was hardly observed.

Based on these results, commercially available edible gelatin, pharmaceutical gelatin, cosmetic gelatin, and pharmaceutical gelatin hydrolyzed peptide are highly reactive with antigen-specific IgE antibodies in blood for patients with gelatin allergy. In many cases, the gelatin must be avoided, whereas the gelatin of the present invention has low reactivity with antigen-specific IgE antibodies and is safe. Also, gummy candy, jelly produced using the gelatin or peptide of the present invention. — And fruit juice beverages can be safely used in gelatin allergy patients.

 In the case of commercially available gelatin hydrolyzed peptides, allergenicity was expected to be reduced due to the reduction in molecular weight, but the effect was insufficient as evident from the above test results. On the other hand, the peptide of the present invention is effective in preventing allergic reactions upon administration, because the peptide of the present invention, which is originally low in allergenicity, is hydrolyzed and reduced in molecular weight. Test example 3

Proof of safety (in vivo study)

 In order to investigate the safety of Gummy candi prepared by the method of Example 2, a load test was performed on mild gelatin allergic patients with the cooperation of medical institutions. Incidentally, the load test was performed according to the method described in "Handbook for Food Allergy Countermeasures, edited by Shuichi Uenogawa et al., 143-150, 1996, Science Forum, Tokyo".

① Target patients

 Four subjects (same number of men and women, ages 1 to 5) showed allergic symptoms due to ingestion of gelatin-containing food. Before the study, the purpose and contents of the study were fully explained, and then the consent was obtained from the legal representative of the patient and, if possible, the patient. Patients who fall under any of the following items were excluded.

 · Patients who have received steroids, antihistamines, antiallergic drugs, etc. that modify the reaction.

 • Patients whose symptoms have not been relieved due to removal of gelatin or have been removed for less than one month. • Patients at risk of anaphylactic shock or acute asthma.

 • Others who are judged inappropriate by the attending physician.

② Test food

Gummy candies prepared according to the method of Example 2 to a size of 2.5 g per grain were used. ③ Administration method

 All patients were kept in the hospital, and always had an intravenous route to prevent anaphylactic shock, and were administered on an empty stomach in the presence of a legal representative. The dose was determined with reference to the results of the specific IgE antibody value for gelatin, and the dose was determined in a single blind manner. The dose was 10 tablets per person, and a small dose (1 tablet) was given at first. If there was no response, the dose was increased at 15-minute intervals, and all doses were completed in 1 hour. If symptoms appeared on the way, the load was stopped at that point and observation was made.

 ④ Observation method

 Observations were divided into skin symptoms, gastrointestinal symptoms, respiratory symptoms, and anaphylaxis, respectively, before administration, at the time of administration, 30 minutes after administration, 1 hour, 2 hours, 3 hours, and 5 hours after administration. After 24 hours, follow-up until 48 hours.

 ⑤Evaluation items

 The physician evaluated the following three points.

 (1) Did the food produce any symptoms or findings (positive diagnosis)?

 (2) Where on the body what symptoms · The appearance of the findings (diagnosis of disease).

 (3) The time from the initial administration until the appearance of symptoms and findings (diagnosis of appearance time).

 ⑥ Judgment

 The physician comprehensively considered the evaluation items at each observation and evaluated the safety of the test food on a three-point scale: “no problem, slight problem, problem”.

⑦ Result

Table 3 shows the observation results of this load test. The overall evaluation result was that 3 out of 4 target patients were “no problem” and one was “somewhat problematic”. In this patient, a commercially available gummy candy was confirmed to have juniper after taking one grain, but a gummy candy that used the gelatin of the present invention had no problem with one grain, and mildly on her back 30 minutes after ingesting 10 grains. Juniper measles was confirmed. Table 3

 Based on the above results, this test food can be said to be a safe and delicious alternative food for gelatin allergy patients.

Claims

The scope of the claims 1. It must be derived from poultry skin, bone and Z or tendon and have an isoelectric point of 7 to 10 and no antigen-antibody reaction with the serum of a patient with gelatin argerium or low antigen-antibody reaction. Particularly, edible, medical or cosmetic gelatin.  2. The edible, medical or cosmetic gelatin according to claim 1, which is a gelatin obtained by acid-treating poultry skin, bone and Z or tendon.  3. A food having low allergenicity for a gelatin allergic patient, comprising the gelatin according to claim 1. 4. A jelly, a gummy candy, a yogurt, a nose, a sausage, a sp, a bavarois or an ice cream, which contains the gelatin according to claim 1 and has low allergenicity for a gelatin allergy patient.  5. A cosmetic product comprising the gelatin according to claim 1, which is hypoallergenic to a patient with gelatin allergy. 6. A cream, ointment or efficiency_ffiK which contains gelatin as described in claim 1 and is hypoallergenic for gelatin-allergic patients.  7. A medical product containing the gelatin described in claim 1 and having low allergenicity for a gelatin allergic patient.  8. A pharmaceutical capsule, poultice, or vaccine having low allergenicity for a gelatin allergic patient, comprising the gelatin according to claim 1.  9. Claims comprising immersing poultry skin, bone and / or tendon in an acid solution, collecting the precipitate, heat-extracting the obtained precipitate with an acid solution, and purifying the extract. 1. The method for producing gelatin described in 1.  10. An edible, medical or cosmetic peptide obtained by hydrolyzing the gelatin according to claim 1 and Z or collagen which is a precursor thereof.